LOCUS Exported 9550 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Lentiviral expression of negative control sgRNA for CRISPRi knockdown. ACCESSION . VERSION . KEYWORDS Lenti_CRISPRi_sgNeg2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9550) AUTHORS Ma M, Ghosh S, Tavernari D, Katarkar A, Clocchiatti A, Mazzeo L, Samarkina A, Epiney J, Yu YR, Ho PC, Levesque MP, Ozdemir BC, Ciriello G, Dummer R, Dotto GP TITLE Sustained androgen receptor signaling is a determinant of melanoma cell growth potential and tumorigenesis. JOURNAL J Exp Med. 2021 Feb 1;218(2). pii: 211509. doi: 10.1084/jem.20201137. PUBMED 33112375 REFERENCE 2 (bases 1 to 9550) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9550 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 13..239 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 240..420 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 467..592 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1085..1318 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1503..1547 /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 1725..1965 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 1725..1745 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 1896..1915 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 1996..2071 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_feature 2127..2244 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2383..3561 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2614..3552 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3509..3529 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 3568..3577 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3583..4599 /codon_start=1 /product="hygromycin B phosphotransferase" /label=HygR /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK" CDS 4624..4680 /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 4681..5397 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4726..4747) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4987..5006) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5334..5355 /label=EGFP-C /note="EGFP, forward primer" misc_feature 5415..6003 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5468..5488) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5886..5897) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6075..6308 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 6386..6507 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6423..6442) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6477..6496 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 6547..6682 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6609..6628 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(6702..6721) /label=T7 /note="T7 promoter, forward primer" promoter complement(6703..6721) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6731..6748) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(6731..6747) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6740..6762) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 6889..7344 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6976..6995) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7186..7207 /label=F1ori-F /note="F1 origin, forward primer" promoter 7370..7474 /gene="bla" /label=AmpR promoter CDS 7475..8335 /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(7693..7712) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8506..9094 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8995..9014 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 9248..9265 /label=L4440 /note="L4440 vector, forward primer" protein_bind 9382..9403 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 9418..9448 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9456..9472 /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 9461..9483 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 9480..9496 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 9480..9496 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 9515..9535 /label=T3 /note="T3 promoter, forward primer" promoter 9517..9535 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga 61 gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa 121 ggtggtacga tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa 181 ccactgaatt gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg 241 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 301 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 361 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 421 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag 481 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc 541 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa 601 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa 661 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc 721 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct 781 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt 841 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca 901 aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag 961 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat 1021 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg 1081 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt 1141 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca 1201 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca 1261 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg 1321 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt 1381 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag 1441 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag 1501 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta 1561 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag 1621 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac 1681 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cagagagggc ctatttccca 1741 tgattccttc atatttgcat atacgataca aggctgttag agagataatt agaattaatt 1801 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg 1861 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt 1921 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accgtgcgat 1981 gggggggtgg gtagcgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta 2041 tcaacttgaa aaagtggcac cgagtcggtg cttttttaag cttggcgtaa ctagatcttg 2101 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt 2161 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa 2221 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccactttgg 2281 cgccggctcg agggggcccg ggtgcaaaga tggataaagt tttaaacaga gaggaatctt 2341 tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag 2401 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga 2461 tccggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc 2521 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt 2581 ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg 2641 cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctgcag 2701 tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc 2761 gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc 2821 cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc 2881 atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat 2941 gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc 3001 cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc 3061 ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt 3121 atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga 3181 tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag 3241 cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca 3301 tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc gagcttttgg 3361 agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc ccacactgag 3421 tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc 3481 ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt 3541 cttccatttc aggtgtcgtg acgtacggcc accatggaat tcaaaaagcc tgaactcacc 3601 gctacctctg tcgagaagtt tctgatcgaa aagtttgaca gtgtatccga cctgatgcag 3661 ctctccgagg gcgaagaatc tcgggctttc agcttcgatg tgggagggcg tggatatgtc 3721 ctgcgggtga atagctgcgc cgatggtttc tacaaagatc gctatgttta tcggcacttt 3781 gcatccgccg ctctccctat tcccgaagtg cttgacattg gggagttcag cgagagcctg 3841 acctattgca tctcccgccg tgcacagggt gtcaccttgc aagacctgcc tgaaaccgaa 3901 ctgcccgctg ttctccagcc cgtcgccgag gccatggatg ccatcgctgc cgccgatctt 3961 agccagacca gcgggttcgg cccattcgga cctcaaggaa tcggtcaata cactacatgg 4021 cgcgatttca tctgcgctat tgctgatccc catgtgtatc actggcaaac tgtgatggac 4081 gacaccgtca gtgcctccgt cgcccaggct ctcgatgagc tgatgctttg ggccgaggac 4141 tgccccgaag tccggcacct cgtgcacgcc gatttcggct ccaacaatgt cctgaccgac 4201 aatggccgca taacagccgt cattgactgg agcgaggcca tgttcgggga ttcccaatac 4261 gaggtcgcca acatcttctt ctggaggccc tggttggctt gtatggagca gcagacccgc 4321 tacttcgagc ggaggcatcc cgagcttgca ggatctcctc ggctccgggc ttatatgctc 4381 cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct 4441 tgggctcagg gtcgctgcga cgcaatcgtc cggtccggag ccgggactgt cgggcgtaca 4501 caaatcgccc gcagaagcgc tgccgtctgg accgatggct gtgtggaagt gctcgccgat 4561 agtggaaaca gacgccccag cactcgtcct agggcaaagg atctgcagtc tagaggatct 4621 ggtgccacga atttctcact actcaagcag gccggtgatg tcgaggaaaa ccctggtcct 4681 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac 4741 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac 4801 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc 4861 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag 4921 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc 4981 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg 5041 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac 5101 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac 5161 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc 5221 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac 5281 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc 5341 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtga 5401 cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc 5461 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg 5521 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc 5581 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg 5641 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg 5701 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga 5761 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct 5821 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg 5881 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc 5941 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc 6001 cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa 6061 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc 6121 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg 6181 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg 6241 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat 6301 ctctagcagt acgtatagta gttcatgtca tcttattatt cagtatttat aacttgcaaa 6361 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata 6421 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 6481 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact 6541 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 6601 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag 6661 tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt 6721 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc 6781 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc 6841 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta 6901 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca 6961 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct 7021 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc 7081 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat 7141 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc 7201 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc 7261 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta 7321 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc 7381 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg 7441 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc 7501 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt 7561 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct 7621 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac 7681 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact 7741 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa 7801 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga 7861 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt 7921 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga 7981 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg 8041 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat 8101 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat 8161 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc 8221 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga 8281 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc 8341 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag 8401 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc 8461 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt 8521 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt 8581 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat 8641 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc 8701 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa 8761 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg 8821 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag 8881 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag 8941 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa 9001 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt 9061 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg 9121 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc 9181 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac 9241 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct 9301 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc 9361 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt 9421 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac 9481 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca 9541 aaagctggag //