LOCUS Exported 11550 bp ds-DNA circular SYN 29-JUL-2021 DEFINITION Tet-regulated (Tet-on) lentiviral vector for transgene (hUbiquitin promoter) - AND/OR - shRNA (H1 promoter when subcloned from pLVTHM (Addgene#12247)) - 3rd generation. ACCESSION . VERSION . KEYWORDS pLVUT-tTR-KRAB SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11550) AUTHORS Szulc J, Wiznerowicz M, Sauvain MO, Trono D, Aebischer P TITLE A versatile tool for conditional gene expression and knockdown. JOURNAL Nat Methods. 2006 Feb . 3(2):109-16. PUBMED 16432520 REFERENCE 2 (bases 1 to 11550) AUTHORS . TITLE Direct Submission JOURNAL Exported Jul 29, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11550 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(42..61) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 233..612 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 614..812 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 767..787 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 830..1010 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1057..1182 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1675..1908 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2093..2137 /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 2435..2552 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2617..3828 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 3665..3684 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" regulatory 3876..3885 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3882..4601 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3882..4598 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3927..3948) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4188..4207) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4535..4556 /label=EGFP-C /note="EGFP, forward primer" misc_feature 4640..5191 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4784..4801) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5011..5030 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5194..5811 /codon_start=1 /gene="tetR from transposon Tn10" /product="tetracycline repressor TetR" /label=TetR /note="TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline." /translation="MARLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG" primer_bind complement(5267..5286) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" CDS 5815..5835 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5938..6132 /codon_start=1 /product="Kruppel-associated box (KRAB) transcriptional repression domain from the human zinc finger protein ZNF10 (Margolin et al., 1994)" /label=KRAB /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY QLTKPDVILRLEKGEEPWLV" misc_feature 6273..6861 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6326..6346) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6744..6755) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(6864..6880) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6865..6881) /label=pBluescriptKS /note="For pBluescript vector" protein_bind 7001..7271 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" protein_bind 7043..7313 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" LTR 7352..7532 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(7556..7573) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 7562..7786 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 7832..8260 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7919..7938) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8129..8150 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(8269..8289) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 8274..8603 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 8454..8589 /label=SV40 ori /note="SV40 origin of replication" primer_bind 8516..8535 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 8651..8698 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 8717..9091 /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 9221..9342 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(9258..9277) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 9312..9331 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(9391..9407) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9391..9407) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(9404..9426) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 9415..9431 /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(9439..9469) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9484..9505 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(9622..9639) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(9793..10381) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(9873..9892) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(10552..11412) /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 11175..11194 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(11413..11517) /gene="bla" /label=AmpR promoter ORIGIN 1 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc 61 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg 121 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt 181 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga 241 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg 301 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc 361 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat 421 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat 481 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat 541 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc 601 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac 661 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa 721 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt 781 aggcgtgtac ggtgggaggt ctatataagc agcgcgtttt gcctgtactg ggtctctctg 841 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 901 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 961 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg 1021 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt 1081 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg 1141 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga 1201 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta 1261 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta 1321 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga 1381 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg 1441 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt 1501 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga 1561 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc 1621 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc 1681 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct 1741 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag 1801 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca 1861 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg 1921 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa 1981 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa 2041 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 2101 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 2161 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 2221 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 2281 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 2341 tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac tgcgtgcgcc 2401 aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg 2461 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat 2521 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc 2581 cagtttggtt aattaacccg tgtcggctcc agatctggcc tccgcgccgg gttttggcgc 2641 ctcccgcggg cgcccccctc ctcacggcga gcgctgccac gtcagacgaa gggcgcagcg 2701 agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg 2761 ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg 2821 cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt 2881 ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg 2941 tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct 3001 gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg 3061 gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg 3121 agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga 3181 gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc 3241 atggtgggcg gcaagaaccc aaggtcttga ggccttcgct aatgcgggaa agctcttatt 3301 cgggtgagat gggctggggc accatctggg gaccctgacg tgaagtttgt cactgactgg 3361 agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc 3421 acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct 3481 tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg 3541 acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga 3601 ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag 3661 tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt 3721 ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact 3781 agtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacga agcttgggct 3841 gcaggtcgac tctagaggat ccccgggtac cggtcgccac catggtgagc aagggcgagg 3901 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca 3961 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt 4021 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct 4081 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt 4141 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact 4201 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga 4261 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca 4321 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca 4381 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca 4441 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg 4501 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg 4561 ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccgc gactctagaa 4621 ttccgccccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg 4681 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa 4741 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg 4801 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca 4861 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc 4921 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt 4981 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg 5041 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca 5101 tgctttacat gtgtttagtc gaggttaaaa aacgtctagg ccccccgaac cacggggacg 5161 tggttttcct ttgaaaaaca cgatgataat accatggcta gattagataa aagtaaagtg 5221 attaacagcg cattagagct gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa 5281 ctcgcccaga agctaggtgt agagcagcct acattgtatt ggcatgtaaa aaataagcgg 5341 gctttgctcg acgccttagc cattgagatg ttagataggc accatactca cttttgccct 5401 ttagaagggg aaagctggca agatttttta cgtaataacg ctaaaagttt tagatgtgct 5461 ttactaagtc atcgcgatgg agcaaaagta catttaggta cacggcctac agaaaaacag 5521 tatgaaactc tcgaaaatca attagccttt ttatgccaac aaggtttttc actagagaat 5581 gcattatatg cactcagcgc tgtggggcat tttactttag gttgcgtatt ggaagatcaa 5641 gagcatcaag tcgctaaaga agaaagggaa acacctacta ctgatagtat gccgccatta 5701 ttacgacaag ctatcgaatt atttgatcac caaggtgcag agccagcctt cttattcggc 5761 cttgaattga tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg gtcgccaaaa 5821 aagaagagaa aggtcgacgg cggtggtgct ttgtctcctc agcactctgc tgtcactcaa 5881 ggaagtatca tcaagaacaa ggagggcatg gatgctaagt cactaactgc ctggtcccgg 5941 acactggtga ccttcaagga tgtatttgtg gacttcacca gggaggagtg gaagctgctg 6001 gacactgctc agcagatcgt gtacagaaat gtgatgctgg agaactataa gaacctggtt 6061 tccttgggtt atcagcttac taagccagat gtgatcctcc ggttggagaa gggagaagag 6121 ccctggctgg tggagagaga aattcaccaa gagacccatc ctgattcaga gactgcattt 6181 gaaatcaaat catcagttta aggatctcga ctagctagta gctagctagc tagtcgagct 6241 caagcttcga attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg 6301 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt 6361 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata 6421 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg 6481 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc 6541 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct 6601 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt 6661 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg 6721 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc 6781 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct 6841 ccctttgggc cgcctccccg catcgatacc gtcgacctcg agtaccttta agaccaatga 6901 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc 6961 taattcactc ccaacgaaga caagatctcg agtttaccac tccctatcag tgatagagaa 7021 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta 7081 ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat 7141 agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg 7201 agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc 7261 agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga 7321 aagtcggatc gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc 7381 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 7441 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 7501 acccttttag tcagtgtgga aaatctctag cagcatctag aaacccgctg atcagcctcg 7561 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc 7621 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt 7681 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat 7741 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa 7801 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg 7861 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 7921 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 7981 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 8041 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 8101 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 8161 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg 8221 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc 8281 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 8341 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 8401 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 8461 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 8521 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt 8581 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg 8641 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa 8701 ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga 8761 cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga 8821 ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga 8881 ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta 8941 cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac 9001 cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg 9061 cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc 9121 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct 9181 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta 9241 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 9301 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc 9361 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 9421 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 9481 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 9541 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 9601 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 9661 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 9721 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 9781 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 9841 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 9901 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 9961 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 10021 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 10081 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 10141 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 10201 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 10261 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 10321 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 10381 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 10441 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 10501 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 10561 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 10621 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 10681 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 10741 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 10801 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 10861 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 10921 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 10981 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 11041 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 11101 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 11161 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 11221 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 11281 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 11341 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 11401 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 11461 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 11521 atttccccga aaagtgccac ctgacgtcga //