LOCUS Exported 9004 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral vector with empty U6 cassette containing LbCpf1 direct repeat and U6 terminator, with constitutive expression of puromycin resistance and Firefly luciferase.. ACCESSION . VERSION . KEYWORDS pRG01-U6-DR-crRNA-BsmbI(x2)-6T; EFS-Puro-2A-Fluc-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9004) AUTHORS Chow RD, Wang G, Ye L, Codina A, Kim HR, Shen L, Dong MB, Errami Y, Chen S TITLE In vivo profiling of metastatic double knockouts through CRISPR-Cpf1 screens. JOURNAL Nat Methods. 2019 May;16(5):405-408. doi: 10.1038/s41592-019-0371-5. Epub 2019 Apr 8. PUBMED 30962622 REFERENCE 2 (bases 1 to 9004) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9004 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 5..231 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 232..412 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 459..584 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1077..1310 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1495..1539 /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" promoter 1717..1957 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 1717..1737 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 1888..1907 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_feature 2069..2186 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2300..2511 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" CDS 2524..3120 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(2524..2543) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 3020..3040 /label=Puro-F /note="Puromycin resistance gene, forward primer" CDS 3130..3186 /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 3187..4839 /codon_start=1 /gene="luc+" /product="firefly luciferase" /label=luciferase /note="enhanced luc+ version of the luciferase gene" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" primer_bind complement(3253..3271) /label=LucNrev /note="Firefly luciferase, reverse primer" misc_feature 4861..5449 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4914..4934) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5332..5343) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5521..5754 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5832..5953 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5869..5888) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5923..5942 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 5993..6128 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6055..6074 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(6148..6167) /label=T7 /note="T7 promoter, forward primer" promoter complement(6149..6167) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6177..6194) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(6177..6193) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6186..6208) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 6335..6790 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6422..6441) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6632..6653 /label=F1ori-F /note="F1 origin, forward primer" promoter 6816..6920 /gene="bla" /label=AmpR promoter CDS 6921..7781 /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(7139..7158) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7952..8540 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8441..8460 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8694..8711 /label=L4440 /note="L4440 vector, forward primer" protein_bind 8828..8849 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 8864..8894 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8902..8918 /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 8907..8929 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 8926..8942 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 8926..8942 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 8961..8981 /label=T3 /note="T3 promoter, forward primer" promoter 8963..8981 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa 61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac 121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa 181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc 241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc 421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc 481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt 541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga 601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat 661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt 721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag 781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa 841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa 901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg 961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta 1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga 1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg 1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg 1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc 1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg 1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat 1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac 1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat 1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca 1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga 1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg 1681 tttcagaccc acctcccaac cccgagggga cccagagagg gcctatttcc catgattcct 1741 tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta 1801 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt 1861 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat 1921 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgtaat ttctactaag 1981 tgtagatgag acggacgtct cttttttaag cttggcgtgg atccgatatc aactagatct 2041 tgagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca 2101 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa 2161 aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga gatccacttt 2221 ggcgccggct cgagggggcc cggggaattc gctagctagg tcttgaaagg agtgggaatt 2281 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 2341 ggaggggtcg gcaattgatc cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 2401 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 2461 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggaccggttc 2521 tagatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc 2581 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg 2641 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc 2701 gacatcggca aggtgtgggt cgcggacgac ggcgccgccg tggcggtctg gaccacgccg 2761 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc 2821 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag 2881 gagcccgcgt ggttcctggc caccgtcgga gtctcgcccg accaccaggg caagggtctg 2941 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc 3001 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc 3061 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc 3121 ggatccggcg caacaaactt ctctctgctg aaacaagccg gagatgtcga agagaatcct 3181 ggaccgatgg aagacgccaa aaacataaag aaaggcccgg cgccattcta tccgctggaa 3241 gatggaaccg ctggagagca actgcataag gctatgaaga gatacgccct ggttcctgga 3301 acaattgctt ttacagatgc acatatcgag gtggacatca cttacgctga gtacttcgaa 3361 atgtccgttc ggttggcaga agctatgaaa cgatatgggc tgaatacaaa tcacagaatc 3421 gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc gttatttatc 3481 ggagttgcag ttgcgcccgc gaacgacatt tataatgaac gtgaattgct caacagtatg 3541 ggcatttcgc agcctaccgt ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac 3601 gtgcaaaaaa agctcccaat catccaaaaa attattatca tggattctaa aacggattac 3661 cagggatttc agtcgatgta cacgttcgtc acatctcatc tacctcccgg ttttaatgaa 3721 tacgattttg tgccagagtc cttcgatagg gacaagacaa ttgcactgat catgaactcc 3781 tctggatcta ctggtctgcc taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga 3841 ttctcgcatg ccagagatcc tatttttggc aatcaaatca ttccggatac tgcgatttta 3901 agtgttgttc cattccatca cggttttgga atgtttacta cactcggata tttgatatgt 3961 ggatttcgag tcgtcttaat gtatagattt gaagaggagc tgtttctgag gagccttcag 4021 gattacaaga ttcaaagtgc gctgctggtg ccaaccctat tctccttctt cgccaaaagc 4081 actctgattg acaaatacga tttatctaat ttacacgaaa ttgcttctgg tggcgctccc 4141 ctctctaagg aagtcgggga agcggttgcc aagaggttcc atctgccagg tatcaggcaa 4201 ggatatgggc tcactgagac tacatcagct attctgatta cacccgaggg ggatgataaa 4261 ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga aggttgtgga tctggatacc 4321 gggaaaacgc tgggcgttaa tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg 4381 tccggttatg taaacaatcc ggaagcgacc aacgccttga ttgacaagga tggatggcta 4441 cattctggag acatagctta ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg 4501 aagtctctga ttaagtacaa aggctatcag gtggctcccg ctgaattgga atccatcttg 4561 ctccaacacc ccaacatctt cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt 4621 gaacttcccg ccgccgttgt tgttttggag cacggaaaga cgatgacgga aaaagagatc 4681 gtggattacg tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt 4741 gtggacgaag taccgaaagg tcttaccgga aaactcgacg caagaaaaat cagagagatc 4801 ctcataaagg ccaagaaggg cggaaagatc gccgtgtaag aattcacgcg ttaagtcgac 4861 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 4921 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 4981 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 5041 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 5101 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 5161 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 5221 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc 5281 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 5341 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt 5401 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa 5461 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 5521 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc 5581 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 5641 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 5701 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtacgt 5761 atagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 5821 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 5881 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 5941 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc 6001 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 6061 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 6121 tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg 6181 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt 6241 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct 6301 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc 6361 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 6421 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 6481 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 6541 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 6601 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 6661 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 6721 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg 6781 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt 6841 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat 6901 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt 6961 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg 7021 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga 7081 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc 7141 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac 7201 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg 7261 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca 7321 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg 7381 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg 7441 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg 7501 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag 7561 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg 7621 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct 7681 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac 7741 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact 7801 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga 7861 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt 7921 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 7981 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 8041 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc 8101 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 8161 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 8221 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 8281 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 8341 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 8401 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 8461 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 8521 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt 8581 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta 8641 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt 8701 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc 8761 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca 8821 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc 8881 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg 8941 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc 9001 aagc //