LOCUS        pLenti-Tet-coBxb1-2A-BFP_IRES-iCasp9-2A-Blast_rtTA3       12668 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentivector encoding a DNA recombinase landing pad with Bxb1 
            integrase and iCasp9 negative selection behind a Tet-inducible 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12668)
  AUTHORS   Shukla N, Roelle SM, Suzart VG, Bruchez AM, Matreyek KA
  TITLE     Mutants of human ACE2 differentially promote SARS-CoV and SARS-CoV-2
            spike mediated infection.
  JOURNAL   PLoS Pathog. 2021 Jul 16;17(7):e1009715. doi: 
            10.1371/journal.ppat.1009715. eCollection 2021 Jul.
  PUBMED    34270613
REFERENCE   2  (bases 1 to 12668)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            //gt12365.cn
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Pathog."; date: "2021-07-16"; volume: "17(7):e1009715. doi"; pages: 
            " 10.1371/journal.ppat.1009715. eCollection 2021 Jul"
COMMENT     Sequence Label: pLenti-Tet-coBxb1-2A-BFP_IRES-iCasp9-2A-Blast_...
FEATURES             Location/Qualifiers
     source          1..12668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(140..166)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(167..187)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    197..244
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     promoter        complement(246..595)
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     protein_bind    353..371
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    389..407
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    425..443
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    461..479
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    497..515
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    533..551
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    569..587
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    738..854
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(1121..1165)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1350..1583
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2076..2201
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             2248..2428
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(2429..2655)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(2683..2703)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2683..2701)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2722..2738)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2722..2738)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2735..2757)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2746..2762
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2770..2776,2777..2794,2795..2800))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2815..2836
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2953..2970)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3124..3712)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3204..3223)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3883..4674,4675..4743))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4506..4525
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4744..4848)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4874..5329)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5011..5032)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5223..5242
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5456..5478
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5470..5487
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5471..5487
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5497..5516
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5497..5515
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      5536..5671
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5711..5832
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5722..5741)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5776..5795
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             5904..6137
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     CDS             complement(6224..6931)
                     /codon_start=1
                     /product="reverse tetracycline transactivator 3"
                     /label=rtTA3
                     /note="activates expression from Tet-responsive promoters 
                     in the presence of doxycycline"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPG"
     protein_bind    complement(6934..6958)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(7063..7265)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        7266..7569
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7885..7904
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    7903..7958
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     8004..8023
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(8116..8511)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(8512..8565)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(8584..8610)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             9124..9135
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             complement(9487..9807)
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    join(9817..9825,9826..9828,9829..10403)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(9990..10009)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     10219..10236
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(10528..11235)
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMR
                     IKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDM
                     ALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVA
                     RYCDLPSKLGHKLN"
     CDS             complement(11236..11295)
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
ORIGIN
        1 cgtccagatc ctctgccacg ccgaccacat cccaaccccg ctgagcacag agctgctggc
       61 atgattccag ctgtctctcg ggacttgtgg tggcgtctgt cactcggctc agtctaatca
      121 cgaccagggc tcttgaaccg gcataatcgg ggacatcata gggatacact tttcttttct
      181 tttttggcat ggtggcggtt tgtaccgtac accactgaga ccgcggtggt tgaccagaca
      241 aacccaagta taagacaaaa gtgttgtgga attgctccag gcgatctgac ggttcactaa
      301 acgagctctg cttttatagg cgcccaccgt acacgcctac ctcgacatac gttctctatc
      361 actgataggg agtaaactgg atatacgttc tctatcactg atagggagta aactgtagat
      421 acgttctcta tcactgatag ggagtaaact ggtcatacgt tctctatcac tgatagggag
      481 taaactcctt atacgttctc tatcactgat agggagtaaa gtctgcatac gttctctatc
      541 actgataggg agtaaactct tcatacgttc tctatcactg atagggagta aactcgagtg
      601 aagacgaaag ggcctcatcg cataagcctt aatatcgata aactggatct ctgctgaaga
      661 cgaaagggcc tcatcgcaat ttataaacat ccgataaact ggatctctgc tgtccctgaa
      721 ctcccaagct tatcgataaa attttgaatt ttgtaatttg tttttgtaat tctttagttt
      781 gtatgtctgt tgctattatg tctactattc tttcccctgc actgtacccc ccaatccccc
      841 cttttctttt aaaagttaac cgataccgtc gagatccgtt cactaatcga atggatctgt
      901 ctctgtctct ctctccacct tcttcttcta ttccttcggg cctgtcgggt cccctcgggg
      961 ttgggaggtg ggtctgaaac gataatggtg aatatccctg cctaactcta ttcactatag
     1021 aaagtacagc aaaaactatt cttaaaccta ccaagcctcc tactatcatt atgaataatt
     1081 ttatatacca cagccaattt gttatgttaa accaattcca caaacttgcc catttatcta
     1141 attccaataa ttcttgttca ttcttttctt gctggttttg cgattcttca attaaggagt
     1201 gtattaagct tgtgtaattg ttaatttctc tgtcccactc catccaggtc gtgtgattcc
     1261 aaatctgttc cagagattta ttactccaac tagcattcca aggcacagca gtggtgcaaa
     1321 tgagttttcc agagcaaccc caaatcccca ggagctgttg atcctttagg tatctttcca
     1381 cagccaggat tcttgcctgg agctgcttga tgccccagac tgtgagttgc aacagatgct
     1441 gttgcgcctc aatagccctc agcaaattgt tctgctgctg cactatacca gacaataatt
     1501 gtctggcctg taccgtcagc gtcattgacg ctgcgcccat agtgcttcct gctgctccca
     1561 agaacccaag gaacaaagct cctattccca ctgctctttt ttctctctgc accactcttc
     1621 tctttgcctt ggtgggtgct actcctaatg gttcaatttt tactacttta tatttatata
     1681 attcacttct ccaattgtcc ctcatatctc ctcctccagg tctgaagatc agcggccgct
     1741 tgctgtgcgg tggtcttact tttgttttgc tcttcctcta tcttgtctaa agcttccttg
     1801 gtgtctttta tctctatcct ttgatgcaca caatagaggg ttgctactgt attatataat
     1861 gatctaagtt cttctgatcc tgtctgaagg gatggttgta gctgtcccag tatttgtcta
     1921 cagccttctg atgtttctaa caggccagga ttaactgcga atcgttctag ctccctgctt
     1981 gcccatacta tatgttttaa tttatatttt ttctttcccc ctggccttaa ccgaattttt
     2041 tcccatcgcg atctaattct cccccgctta atactgacgc tctcgcaccc atctctctcc
     2101 ttctagcctc cgctagtcaa aatttttggc gtactcacca gtcgccgccc ctcgcctctt
     2161 gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga gctcctctgg tttccctttc
     2221 gctttcaagt ccctgttcgg gcgccactgc tagagatttt ccacactgac taaaagggtc
     2281 tgagggatct ctagttacca gagtcacaca acagacgggc acacactact tgaagcactc
     2341 aaggcaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag agctcccagg
     2401 ctcagatctg gtctaaccag agagacccgt ttatgtatcg agctaggcac ttaaatacaa
     2461 tatctctgca atgcggcaat tcagtggttc gtccaatcca tgtcagaccc gtctgttgcc
     2521 ttcctaataa ggcacgatcg taccacctta cttccaccaa tcggcatgca cggtgctttt
     2581 tctctccttg taaggcatgt tgctaactca tcgttaccat gttgcaagac tacaagagta
     2641 ttgcataaga ctacattaag cttgcagctc cagcttttgt tccctttagt gagggttaat
     2701 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2761 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     2821 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2881 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2941 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3001 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3061 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3121 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3181 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3241 gcgctctcct gttccgaccc tgccgctcac cggatacctg tccgcctttc tcccttcggg
     3301 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3361 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3421 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3481 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3541 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3601 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3661 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3721 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3781 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3841 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3901 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3961 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4021 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4081 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4141 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4201 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4261 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4321 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4381 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4441 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4501 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4561 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4621 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4681 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4741 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4801 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4861 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     4921 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     4981 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     5041 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     5101 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     5161 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     5221 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     5281 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
     5341 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     5401 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     5461 tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa
     5521 ttgggtacgt ccctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
     5581 ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat
     5641 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg atagctagag
     5701 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     5761 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     5821 aataaacaag ttcctctcac tctctgatat tcatttcttt gcaagttata aatactgaat
     5881 aataagatga catgaactac tactgctaga gattttccac actgactaaa agggtctgag
     5941 ggatctctag ttaccagagt cacacaacag acgggcacac actacttgaa gcactcaagg
     6001 caagctttat tgaggcttaa gcagtgggtt ccctagttag ccagagagct cccaggctca
     6061 gatctggtct aaccagagag acccagtaca agcaaaaagc agatcttgtc ttcgttggga
     6121 gtgagctagc ccttccagtc cccccttttc ttttaaaaag tggctaagat ctacagctgc
     6181 cttgtaagtc attggtctta aaggtaccga gctcgaattg tgcttacccg gggagcatgt
     6241 caaggtcaaa atcgtcaagg gcatcggctg ggagcatgtc taagtcaaaa tcgtcaaggg
     6301 cgtcggtcgg cccgccgctt tcgcacttta gctgtttctc caggccacat atgattagtt
     6361 ccaggccgaa aaggaaggca ggttcggctc cctgccggtc gaacagctca attgcttgtc
     6421 tcagaagtgg gggcatagaa tcggtggtag gtgtctctct ttcctctttt gctacttgat
     6481 gctcctgttc ctccaatacg cagcccagtg taaagtggcc cacggcggac agagcgtaca
     6541 gtgcgttctc cagggagaag ccttgctgac acaggaacgc gagctgattt tccagggttt
     6601 cgtactgttt ctctgttggg cgggtgccga gatgcacttt agccccgtcg cgatgtgaga
     6661 ggagagcaca gcggtatgac ttggcgttgt tccgcagaaa gtcttgccat gactcgcctt
     6721 ccagggggca gaagtgggta tgatgcctgt ccagcatctc gattggcagg gcatcgagca
     6781 gggcccgctt gttcttcacg tgccagtaca gggtaggctg ctcaactccc agcttttgag
     6841 cgagtttcct tgtcgtcagg ccttcgatac cgacaccatt gagtaattcc agagctccgt
     6901 ttatgacttt gctcttgtcc agtctagaca tggtgccaac ttttttgtac aaacttgttg
     6961 atatctgcag aattccacca cactggacta gagtcggtgt cttctatgga ggtcaaaaca
     7021 gcgtggatgg cgtctccagg cgatctgacg gttcactaaa cgagctctgc ttatatagac
     7081 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     7141 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     7201 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaccgc
     7261 atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat
     7321 aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg
     7381 ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac
     7441 tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt
     7501 attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt
     7561 tatgtaacgc ggtttagatc tccataagag aagagggaca gctatgactg ggagtagtca
     7621 ggagaggagg aaaaatctgg ctagtaaaac atgtaaggaa aattttaggg atgttaaaga
     7681 aaaaaataac acaaaacaaa atataaaaaa aatctaacct caagtcaagg cttttctatg
     7741 gaataaggaa tggacagcag ggggctgttt catatactga tgacctcttt atagccaacc
     7801 tttgttcatg gcagccagca tatgggcata tgttgccaaa ctctaaacca aatactcatt
     7861 ctgatgtttt aaatgatttg ccctcccata tgtccttccg agtgagagac acaaaaaatt
     7921 ccaacacact attgcaatga aaataaattt cctttattag ccagaagtca gatgctcaag
     7981 gggcttcatg atgtccccat aatttttggc agagggaaaa agatctcagt ggtatttgtg
     8041 agccagggca ttggccacac cagccaccac cttctgatag gcagcctgca cctgaggagt
     8101 gaattcacgc gtttactagc cctcccacac ataaccagag ggcagcaatt cacgaatccc
     8161 aactgccgtc ggctgtccat cactgtcctt cactatggct ttgatcccag gatgcagatc
     8221 gagaagcacc tgtcggcacc gtccgcaggg gctcaagatg cccctgttct catttccgat
     8281 cgcgacgata caagtcaggt tgccagctgc cgcagcagca gcagtgccca gcaccacgag
     8341 ttctgcacaa ggtcccccag taaaatgata tacattgaca ccagtgaaga tgcggccgtc
     8401 gctagagaga gctgcgctgg cgacgctgta gtcttcagag atggggatgc tgttgattgt
     8461 agccgttgct ctttcaatga gggtggattc ttcttgagac aaaggcttgg cagggccggg
     8521 attctcctcc acgtcaccgc atgttagaag acttcctctg ccctctccgc tgccagagtc
     8581 gagtgcgtag tctggtacgt cgtacggata gtcgactgat gttttaaaga aaagtttttt
     8641 ccggaggaaa ttaaagcaac caggcatctg tttataaatc cctttcaccg aaacagcatt
     8701 agcgacccta agcaggaggg actgcaggtc ttcagagtga gcccactgct caaagatgtc
     8761 gtccagggtc tcaacgtacc aggagccact cttggggtcc ctccaggaaa caaaacctgg
     8821 gaaagtagag taggacacaa agatgtcact gggtgtgggc aaactagata tggcgtccag
     8881 ctggtcgaag gtcctcaaac cttcctggaa cggggtggca tctggctcgg ggttactgcc
     8941 aggggactcg tcttcagggg aagtggaggc cacctcaaac ccatggtctt tctgctcccc
     9001 accacaggcc tggatgaaaa agagcttggg cttccctccc aggctggggc agctggtccc
     9061 attgaagatg ttcacaatct tctcgaccga cacagggcat ccatctgtgc cgtagacagc
     9121 ccctgggaac tgcaggtggc tggcctgaca gccgtgagag agaatgacca ccacgcagca
     9181 gtccagagca ccgtggtcct gccgcgccag ctccagcaaa gccagcacca ttttcttggc
     9241 agtcaggtcg cccttcacct ccaccatgaa atgcagcgag gagaagcgac gccgcaactt
     9301 ctcacagtcg atgttggagc cagtgcgggt gcggagcccg gactcacggc agaagttcac
     9361 attgttgata atgaggcagt ggccacaggg ctccatgctc aggatgtaag ccaaatctgc
     9421 atttcccctc aaactctcaa gagcaccgac atcaccaaat ccgtcgactc cggatccacc
     9481 gccagattcc agttttagaa gctccacatc gaagacgaga gtggcatgtg gtgggatgat
     9541 gcctgggtgc ccagtggcac cataggcata atctggagat atagtcagtt tggctctctg
     9601 acccacactc atctgggcaa ccccttcttc ccagcctcgg atcacctcct gcttgcctag
     9661 cataaactta aagggcttgt ttctgtcccg ggaggaatca actttctttc catcttcaag
     9721 catcccggtg tagtgcacca cgcaggtctg gccgcgcttg gggaaggtgc gcccgtctcc
     9781 tggggagata gtctccacct gcactccctc gagcatggtt gtggccatat tatcatcgtg
     9841 tttttcaaag gaaaaccacg tccccgtggt tcggggggcc tagacgtttt tttaacctcg
     9901 actaaacaca tgtaaagcat gtgcaccgag gccccagatc agatcccata caatggggta
     9961 ccttctgggc atccttcagc cccttgttga atacgcttga ggagagccat ttgactcttt
    10021 ccacaactat ccaactcaca acgtggcact ggggttgtgc cgcctttgca ggtgtatctt
    10081 atacacgtgg cttttggccg cagaggcacc tgtcgccagg tggggggttc cgctgcctgc
    10141 aaagggtcgc tacagacgtt gtttgtcttc aagaagcttc cagaggaact gcttccttca
    10201 cgacattcaa cagaccttgc attcctttgg cgagagggga aagaccccta ggaatgctcg
    10261 tcaagaagac agggccaggt ttccgggccc tcacattgcc aaaagacggc aatatggtgg
    10321 aaaataacat atagacaaac gcacaccggc cttattccaa gcggcttcgg ccagtaacgt
    10381 tagggggggg agagggagag gggcagtctc tctgattcca cagtgatcag ttatctagat
    10441 ccggtggatc ccgggcccgc ggtaccgtcg actgcagacc tgaagagagg gagaaggagg
    10501 tcagaccggt gaattcttaa gttactaatt aagcttgtgc cccagtttgc tagggaggtc
    10561 gcagtatctg gccactgcca cctcgtgctg ctcgacgtag gtctcgttgt tggcctcctt
    10621 gattctttcc agtctgtagt ccacatagta gacgccaggc atcttgaggt tcttagcggg
    10681 tttcttggat ctatatgtgg tcttggcgtt tgcgatcaga tggctcccgc ccacgagctt
    10741 cagggccatg tcgtttctgc cttccaggcc gccgtcagcg gggtacagcg tctcggtgaa
    10801 ggcctcccag ccgagtgttt tcttctgcat cacagggccg ttggatgtga agttcacccc
    10861 tctgatcttg acgttgtaga tgaggcagcc gtcctggagg ctggtgtcct gggtagcggt
    10921 cagcacgccc ccgtcttcgt atgtggtgac tctctcccat gtgaagccct cagggaagga
    10981 ctgcttgaag aagtcgggga tgccctgggt gtggttgatg aaggtcttgc tgccgtagag
    11041 gaagctagta gccaggatgt cgaaggcgaa ggggagaggg ccgccctcga ccaccttgat
    11101 tctcatggtc tgggtgccct cgtagggctt gccttcgccc tcggatgtgc acttgaagtg
    11161 atggttgtcc acggtgccct ccatgtacag cttcatgtgc atgttctcct taatcagctc
    11221 ttcgccctta gacacgggac ctgggttgct ctcaacatct ccagccaatt tcaagagagc
    11281 ataattagta cactggcccg agccggacat cccagtgtac agtctttcga ccacggaccc
    11341 cagtctcagg tgctgttcat actcctgcag gtcgccaaaa tcgatggttc gagtcaggcc
    11401 accgcgcacg tcgaatgtga gccggacatt catggatctc agccatgtgt tcttggctgc
    11461 ggtgtcctgc tccctccacc aatccccaaa cctctgtccg gtttctcgcc actcccatcc
    11521 agagggcctg gcctccaggc cctccagttc ctcctgtcta gcagcgagtg cagcaattcg
    11581 tgcgtccaga gcttctctct gtggggaccc agctctatat gcaggagatc cgatgaggga
    11641 agtcagatcc acgagctcgg cattgacttc tgccagctcc acagcggagt cagatcctgc
    11701 cacccacact ttttccagcc gctcagcgtc acccaggaga tccagcacct gttcctcgca
    11761 aaaggcgtcc cactcagcca tggccactgt cccgtttcca cagtgcttag ggaaccccat
    11821 gctcctgcat cgatagcggg gatgtttgcg ccctccgccg gcaaacttgt atgctggctc
    11881 tccacagacg gcgcagaaca gcaccctcag gagcaggctt ggggtgctca cggcaggttt
    11941 ggcgcgacta gtcttgacca gttctgcccg cagagcttcg agctgctctc tggtgagaat
    12001 aggctcagcc ctcaccagag gggctccgtc atcgtcccgg acagttttgc cattgagtgt
    12061 tgcgtaaccc agcatagcct cactgatcat agatctcttc agtgcagtag cactccattc
    12121 tcttccctgt ggctctcttc cctgcagctg agcaaaataa tcctttggag agagcacgcc
    12181 gcgtctattc aggtcgtgtg ccaccagatg gagtggttcg tggttatcga ccactctgtg
    12241 gtacacttcc agaatccgct ctctctggac ggggtctggc accagtctcc actccccatc
    12301 caccctggta gggagatatc cccaaggggg cagggagcct ctgtatttac cggccctaat
    12361 attgaaatgg gctgcgctcc tgtttcgctc cttgatagct tcgagctcca tctgggccac
    12421 agtgcccatc agtgcgatga ccacagcggc aaaaggtgtg gtagtatcga agtgggcttc
    12481 tgttgcgctg accaccagtt tcttgtggtc ctcagcccaa tgcaccagct gctggagatg
    12541 tcgaattgag cgggtcagcc ggtccactct gtaggcgacg atcacatcaa agggctgttc
    12601 ctcgaaagcg agccatcttg ccaggttggg cctcctcttt ctgtcgaatg gatcgacggc
    12661 cccgctca
//