LOCUS       lenti-sgRNA blast       10437 bp    DNA     circular SYN 14-APR-2025
DEFINITION  This 3rd generation lentiviral plasmid expresses a S. pyogenes 
            CRISPR chimeric RNA element with customizable sgRNA from a U6 
            promoter and blasticidin S resistance from an EF-1a promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10437)
  AUTHORS   Stringer BW, Day BW, D'Souza RCJ, Jamieson PR, Ensbey KS, Bruce ZC, 
            Lim YC, Goasdoue K, Offenhauser C, Akgul S, Allan S, Robertson T, 
            Lucas P, Tollesson G, Campbell S, Winter C, Do H, Dobrovic A, Inglis
            PL, Jeffree RL, Johns TG, Boyd AW
  TITLE     A reference collection of patient-derived cell line and xenograft 
            models of proneural, classical and mesenchymal glioblastoma.
  JOURNAL   Sci Rep. 2019 Mar 20;9(1):4902. doi: 10.1038/s41598-019-41277-z.
  PUBMED    30894629
REFERENCE   2  (bases 1 to 10437)
  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: "Sci Rep."; 
            date: "2019-03-20"; volume: "9(1):4902. doi"; pages: " 
            10.1038/s41598-019-41277-z"
COMMENT     Sequence Label: lenti-sgRNA blast
FEATURES             Location/Qualifiers
     source          1..10437
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..30
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(149..169)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        154..483
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      334..469
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     396..415
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        531..578
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             597..971
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    1101..1222
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1138..1157)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1192..1211
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1271..1287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1271..1287)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1284..1306)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1295..1311
                     /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(1319..1325,1326..1343,1344..1349))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1364..1385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1502..1519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1673..2261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1753..1772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2432..3223,3224..3292))
                     /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     3055..3074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3293..3397)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3472..3491)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3667..4046
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4048..4246
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4201..4221
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4264..4444
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4491..4616
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5109..5342
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5527..5571
                     /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    5869..5986
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6037..6285
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6037..6057
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6208..6227
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8167..8242
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8304..8515
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      8534..8543
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8540..8938
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8954..9542
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9007..9027)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9425..9436)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9614..9847
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9873..9890)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9879..10103
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(10149..10437,1..140)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10236..10255)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
       61 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      121 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
      181 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
      241 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
      301 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
      361 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
      421 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
      481 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
      541 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
      601 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
      661 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
      721 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
      781 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
      841 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
      901 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
      961 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
     1021 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     1081 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     1141 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1201 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     1261 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     1321 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     1381 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     1441 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     1501 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1561 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1621 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1681 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1741 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1801 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1861 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1921 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1981 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2041 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2101 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2161 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2221 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2281 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2341 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2401 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2461 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     2521 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     2581 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     2641 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     2701 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     2761 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     2821 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     2881 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     2941 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3001 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3061 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3121 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3181 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3241 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3301 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3361 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     3421 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     3481 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     3541 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     3601 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
     3661 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     3721 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     3781 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     3841 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     3901 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     3961 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     4021 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     4081 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     4141 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     4201 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
     4261 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     4321 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     4381 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     4441 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     4501 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     4561 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     4621 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     4681 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     4741 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     4801 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     4861 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     4921 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
     4981 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     5041 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     5101 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     5161 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     5221 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     5281 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     5341 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     5401 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     5461 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     5521 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     5581 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     5641 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     5701 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     5761 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg
     5821 gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa
     5881 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     5941 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     6001 ggacagcaga gatccagttt ggttaattaa ggtaccgagg gcctatttcc catgattcct
     6061 tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta
     6121 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
     6181 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
     6241 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggaga cggttgtaaa
     6301 tgagcacaca aaatacacat gctaaaatat tatattctat gacctttata aaatcaacca
     6361 aaatcttctt tttaataact ttagtatcaa taattagaat ttttatgttc ctttttgcaa
     6421 acttttaata aaaatgagca aaataaaaaa acgctagttt tagtaactcg cgttgttttc
     6481 ttcaccttta ataatagcta ctccaccact tgttcctaag cggtcagctc ctgcttcaat
     6541 cattttttga gcatcttcaa atgttctaac tccaccagct gctttaacta aagcattgtc
     6601 tttaacaact gacttcatta gtttaacatc ttcaaatgtt gcacctgatt ttgaaaatcc
     6661 tgttgatgtt ttaacaaatt ctaatccagc ttcaacagct atttcacaag ctttcatgat
     6721 ttcttctttt gttaataaac aattttccat aatacattta acaacatgtg atccagctgc
     6781 tttttttaca gctttcatgt cttctaaaac taattcataa tttttgtctt ttaatgcacc
     6841 aatatttaat accatatcaa tttctgttgc accatcttta attgcttcag aaacttcgaa
     6901 tgcttttgta gctgttgtgc atgcacctag aggaaaacct acaacatttg ttattcctac
     6961 atttgtgcct tttaataatt ctttacaata gcttgttcaa tatgaattaa cacaaactgt
     7021 tgcaaaatca aattcaattg cttcatcaca taattgttta atttcagctt tcgtagcatc
     7081 ttgttttaat aatgtgtgat ctatatattt gtttagtttc attttttctc ctatatattc
     7141 atttttaatt ttaattcttt aataatttcg tctactttaa ctttagcgtt ttgaacagat
     7201 tcaccaacac ctataaaata aatttttagt ttaggttcag ttccacttgg gcgaacagca
     7261 aatcatgact tatcttctaa ataaaatttt agtaagtctt gtcctggcat attatacatt
     7321 ccatcgatgt agtcttcaac attaacaact ttaagtccag caatttgagt taagggtgtt
     7381 gctctcaatg atttcattaa tggttcaatt tttaatttct tttcttctgg tttaaaattc
     7441 aagtttaaag tgaaagtgta atatgcaccc atttctttaa ataaatcttc taaatagtct
     7501 actaatgttt tattttgttt tttataaaat caagcagcct ctgctattaa tatagaagct
     7561 tgtattccat ctttatctct agctgagtca tcaattacat atccataact ttcttcataa
     7621 gcaaaaacaa aatttaatcc gttatcttct tctttagcaa tttctctacc cattcattta
     7681 aatccagtta aagtttttac aatattaact ccatattttt catgagcgat tctatcaccc
     7741 aaatcacttg ttacaaaact tgaatataga gccggatttt ttggaatgct atttaagcgt
     7801 tttagatttg ataattttca atcaattaaa attggtcctg tttgatttcc atctaatctt
     7861 acaaaatgac catcatgttt tattgccatt ccaaatctgt cagcatctgg gtcattcata
     7921 ataataatat ctgcatcatg tttaatacca tattcaagcg gtatttttca tgcaggatca
     7981 aattctggat ttggatttac aacattttta aatgtttcat cttcaaatgc atgctcttca
     8041 acctcaataa cgttatatcc tgattcacgt aatatttttg gggtaaattt agttcctgtt
     8101 ccattaactg cgctaaaaat aatttttaaa tcttttttag cttcttgctc ttttttgtac
     8161 gtctctgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
     8221 aaaagtggca ccgagtcggt gcttttttga attcgctagc taggtcttga aaggagtggg
     8281 aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     8341 ggggggaggg gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg taaactggga
     8401 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     8461 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggaccg
     8521 gttctagagc gctgccacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga
     8581 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
     8641 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
     8701 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
     8761 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
     8821 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
     8881 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaaac
     8941 gcgttaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     9001 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     9061 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     9121 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     9181 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     9241 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     9301 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     9361 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     9421 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     9481 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     9541 gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     9601 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
     9661 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     9721 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     9781 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     9841 tctagcaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     9901 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     9961 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
    10021 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
    10081 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
    10141 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
    10201 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
    10261 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
    10321 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
    10381 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgtt
//