LOCUS       pLenti-PGK-Neo-PIP-FUCCI       10145 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentiviral expression of the cell cycle marker PIP-FUCCI.
ACCESSION   .
VERSION     .
KEYWORDS    .pLenti-PGK-Neo-PIP-FUCCI
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10145)
  AUTHORS   Grant GD, Kedziora KM, Limas JC, Cook JG, Purvis JE
  TITLE     Accurate delineation of cell cycle phase transitions in living cells
            with PIP-FUCCI.
  JOURNAL   Cell Cycle. 2018;17(21-22):2496-2516. doi: 
            10.1080/15384101.2018.1547001.
  PUBMED    30421640
REFERENCE   2  (bases 1 to 10145)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            //www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell 
            Cycle."; date: "2018"; volume: "17(21-22):2496-2516. doi"; pages: " 
            10.1080/15384101.2018.1547001"
COMMENT     Sequence Label: pLenti-PGK-Neo-PIP-FUCCI
FEATURES             Location/Qualifiers
     source          1..10145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /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    1575..1692
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1738..2248
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2167..2185
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     protein_bind    2303..2327
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2399..2419
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2429..2455
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2459..2485
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(2492..2494,2495..2497,2498..3205)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(2537..2558)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2798..2817)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3145..3166
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3221..3277
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3284..3991
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3429..3447)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3697..3716
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3998..4327
                     /codon_start=1
                     /product="degron consisting of residues 1-110 of human 
                     Geminin (Zielke and Edgar, 2015)"
                     /label=Gem1 (1-110)
                     /translation="MNPSMKQKQEEIKENIKNSSVPRRTLKMIQPSASGSLVGRENELS
                     AGLSKRKHRNDHLTSTTSSPGVIVPESSENKNLGGVTQESFDLMIKENPSSQYWKEVAE
                     KRRKAL"
     protein_bind    complement(4344..4368)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4399..4987
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4452..4472)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4870..4881)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        5030..5529
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5065..5088)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5462..5481
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             5543..6346
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5606..5625)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6216..6235
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             6437..6670
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6742..6863
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6779..6798)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6833..6852
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6903..7038
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7058..7077)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7059..7077)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7087..7104)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7087..7103)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7096..7118)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7245..7700
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7332..7351)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7542..7563
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7726..7830
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7831..7899,7900..8691)
                     /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     complement(8049..8068)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8862..9450
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9351..9370
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9604..9621
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9738..9759
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9774..9779,9780..9797,9798..9804)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9812..9828
                     /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)."
     primer_bind     9817..9839
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9836..9852
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9836..9852
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9871..9891
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9873..9891
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9919..10145
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     1501 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     1561 cggtatcggt taacttttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     1621 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     1681 attcaaaatt ttatcgatca cgagactagc ctcgagaagc ttgatatcga attccacggg
     1741 gttggggttg cgccttttcc aaggcagccc tgggtttgcg cagggacgcg gctgctctgg
     1801 gcgtggttcc gggaaacgca gcggcgccga ccctgggtct cgcacattct tcacgtccgt
     1861 tcgcagcgtc acccggatct tcgccgctac ccttgtgggc cccccggcga cgcttcctgc
     1921 tccgccccta agtcgggaag gttccttgcg gttcgcggcg tgccggacgt gacaaacgga
     1981 agccgcacgt ctcactagta ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg
     2041 ccgaccgcga tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg
     2101 ccgggaaggg gcggtgcggg aggcggggtg tggggcggta gtgtgggccc tgttcctgcc
     2161 cgcgcggtgt tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt
     2221 tgaccgaatc accgacctct ctccccaggg ggatccaccg gttcgtcgac tagtccagtg
     2281 tggtggaatt ctgcagatat caacaagttt gtacaaaaaa gcaggcttcg aattcatgga
     2341 gcagcgccgc gtcaccgact tcttcgcgcg ccgccgcccc gggccccgga cacgtggccc
     2401 aaaaaagaag agaaaggtag gagctcaata cccatacgat gttcctgact atgcgggcta
     2461 tccctatgac gtcccggact atgcaactag tatggtgagc aagggcgagg agctgttcac
     2521 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2581 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac
     2641 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcctgca
     2701 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2761 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2821 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2881 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     2941 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
     3001 caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     3061 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa
     3121 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     3181 cactctcggc atggacgagc tgtacctcga gggaagcgga gctactaact tcagcctgct
     3241 gaagcaggct ggagacgtgg aggagaaccc tggacctcaa ttgatggtga gcaagggcga
     3301 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     3361 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     3421 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     3481 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     3541 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     3601 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     3661 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     3721 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     3781 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     3841 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     3901 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     3961 ctccaccggc ggcatggacg agctgtacaa ggtcgagatg aatcccagta tgaagcagaa
     4021 acaagaagaa atcaaagaga atataaagaa tagttctgtc ccaagaagaa ctctgaagat
     4081 gattcagcct tctgcatctg gatctcttgt tggaagagaa aatgagctgt ccgcaggctt
     4141 gtccaaaagg aaacatcgga atgaccactt aacatctaca acttccagcc ctggggttat
     4201 tgtcccagaa tctagtgaaa ataaaaatct tggaggagtc acccaggagt catttgatct
     4261 tatgattaaa gaaaatccat cctctcagta ttggaaggaa gtggcagaaa aacggagaaa
     4321 ggcgctgtaa caattgggat cctacccagc tttcttgtac aaagtggttg atatccagca
     4381 cagtggcggc cgctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     4441 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     4501 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     4561 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     4621 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     4681 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     4741 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     4801 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     4861 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     4921 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     4981 tccccgcctg gaattctgca gatatccgga ctagtgatct aattctaccg ggtaggggag
     5041 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc
     5101 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct
     5161 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc
     5221 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt
     5281 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc
     5341 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg
     5401 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg
     5461 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg
     5521 ggcctttcga cctgcagcca atatgggatc ggccattgaa caagatggat tgcacgcagg
     5581 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5641 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5701 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     5761 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5821 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5881 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5941 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     6001 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     6061 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     6121 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     6181 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     6241 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     6301 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagggg atccgtcgag
     6361 ggcccggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     6421 aaagaaaagg ggggactgga agggctagct cactcccaac gaagacaaga tctgcttttt
     6481 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     6541 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     6601 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     6661 atctctagca gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
     6721 tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc
     6781 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     6841 tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc
     6901 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6961 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     7021 gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc
     7081 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     7141 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     7201 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     7261 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     7321 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     7381 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     7441 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     7501 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     7561 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     7621 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     7681 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     7741 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     7801 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     7861 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     7921 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     7981 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     8041 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     8101 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     8161 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     8221 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     8281 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     8341 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     8401 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     8461 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     8521 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     8581 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     8641 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     8701 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     8761 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     8821 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     8881 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     8941 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     9001 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     9061 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     9121 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     9181 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     9241 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     9301 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     9361 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     9421 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     9481 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     9541 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     9601 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     9661 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     9721 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     9781 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     9841 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
     9901 ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa
     9961 cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg
    10021 aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg
    10081 gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca
    10141 taaac
//