LOCUS Exported 9549 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Lentiviral expression of negative control sgRNA for CRISPRi knockdown. ACCESSION . VERSION . KEYWORDS Lenti_CRISPRi_sgNeg1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9549) 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 9549) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9549 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 72..1250 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 303..1241 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1198..1218 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 1257..1266 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1272..2288 /codon_start=1 /product="hygromycin B phosphotransferase" /label=HygR /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK" CDS 2313..2369 /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 2370..3086 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2415..2436) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2676..2695) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3023..3044 /label=EGFP-C /note="EGFP, forward primer" misc_feature 3104..3692 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3157..3177) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3575..3586) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 3764..3997 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4075..4196 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4112..4131) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4166..4185 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 4236..4371 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4298..4317 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(4391..4410) /label=T7 /note="T7 promoter, forward primer" promoter complement(4392..4410) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4420..4437) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4420..4436) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4429..4451) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4578..5033 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4665..4684) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4875..4896 /label=F1ori-F /note="F1 origin, forward primer" promoter 5059..5163 /gene="bla" /label=AmpR promoter CDS 5164..6024 /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(5382..5401) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6195..6783 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6684..6703 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6937..6954 /label=L4440 /note="L4440 vector, forward primer" protein_bind 7071..7092 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7107..7137 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7145..7161 /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 7150..7172 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7169..7185 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7169..7185 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 7204..7224 /label=T3 /note="T3 promoter, forward primer" promoter 7206..7224 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 7252..7478 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 7479..7659 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 7706..7831 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 8324..8557 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 8742..8786 /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 8964..9204 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 8964..8984 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 9135..9154 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 9234..9309 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_feature 9365..9482 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" ORIGIN 1 ggataaagtt ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag 61 gagtgggaat tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg 121 agaagttggg gggaggggtc ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa 181 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt 241 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac 301 aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg 361 tgccttgaat tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg 421 gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag 481 ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct 541 gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc 601 tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg 661 tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc 721 ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg 781 ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc 841 ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct 901 caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa 961 gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca 1021 ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt 1081 tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc 1141 acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca 1201 agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtga cgtacggcca 1261 ccatggaatt caaaaagcct gaactcaccg ctacctctgt cgagaagttt ctgatcgaaa 1321 agtttgacag tgtatccgac ctgatgcagc tctccgaggg cgaagaatct cgggctttca 1381 gcttcgatgt gggagggcgt ggatatgtcc tgcgggtgaa tagctgcgcc gatggtttct 1441 acaaagatcg ctatgtttat cggcactttg catccgccgc tctccctatt cccgaagtgc 1501 ttgacattgg ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg 1561 tcaccttgca agacctgcct gaaaccgaac tgcccgctgt tctccagccc gtcgccgagg 1621 ccatggatgc catcgctgcc gccgatctta gccagaccag cgggttcggc ccattcggac 1681 ctcaaggaat cggtcaatac actacatggc gcgatttcat ctgcgctatt gctgatcccc 1741 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcctccgtc gcccaggctc 1801 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgccg 1861 atttcggctc caacaatgtc ctgaccgaca atggccgcat aacagccgtc attgactgga 1921 gcgaggccat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccct 1981 ggttggcttg tatggagcag cagacccgct acttcgagcg gaggcatccc gagcttgcag 2041 gatctcctcg gctccgggct tatatgctcc gcattggtct tgaccaactc tatcagagct 2101 tggttgacgg caatttcgat gatgcagctt gggctcaggg tcgctgcgac gcaatcgtcc 2161 ggtccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgct gccgtctgga 2221 ccgatggctg tgtggaagtg ctcgccgata gtggaaacag acgccccagc actcgtccta 2281 gggcaaagga tctgcagtct agaggatctg gtgccacgaa tttctcacta ctcaagcagg 2341 ccggtgatgt cgaggaaaac cctggtccta tggtgagcaa gggcgaggag ctgttcaccg 2401 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt 2461 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca 2521 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt 2581 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg 2641 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg 2701 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact 2761 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg 2821 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca 2881 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg 2941 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag 3001 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca 3061 ctctcggcat ggacgagctg tacaagtgac gcgttaagtc gacaatcaac ctctggatta 3121 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg 3181 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc 3241 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca 3301 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac 3361 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact 3421 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc 3481 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg 3541 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc 3601 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac 3661 gagtcggatc tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag 3721 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac 3781 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc 3841 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg 3901 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc 3961 ctcagaccct tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat 4021 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg 4081 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 4141 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat 4201 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 4261 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 4321 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt 4381 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg 4441 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt 4501 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag 4561 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 4621 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt 4681 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc 4741 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga 4801 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 4861 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt 4921 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct 4981 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc 5041 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat 5101 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag 5161 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt 5221 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt 5281 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc 5341 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta 5401 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac 5461 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa 5521 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg 5581 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc 5641 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg 5701 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta 5761 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg 5821 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg 5881 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc 5941 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt 6001 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt 6061 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc 6121 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag 6181 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 6241 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 6301 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag 6361 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 6421 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 6481 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 6541 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 6601 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 6661 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 6721 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 6781 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac 6841 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga 6901 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg 6961 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc 7021 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt 7081 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt 7141 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag 7201 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt 7261 atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag 7321 gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat 7381 taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca 7441 gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga 7501 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct 7561 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat 7621 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt 7681 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg 7741 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg 7801 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg 7861 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt 7921 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga 7981 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact 8041 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa 8101 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc 8161 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa 8221 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg 8281 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg 8341 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg 8401 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg 8461 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc 8521 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa 8581 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac 8641 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct 8701 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat 8761 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt 8821 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg 8881 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc 8941 tcccaacccc gaggggaccc agagagggcc tatttcccat gattccttca tatttgcata 9001 tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat 9061 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat 9121 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg 9181 ctttatatat cttgtggaaa ggacgaaaca ccgcgccaaa cgtgccctga cgggttttag 9241 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg 9301 agtcggtgct tttttaagct tggcgtaact agatcttgag acaaatggca gtattcatcc 9361 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca 9421 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt 9481 ttcgggttta ttacagggac agcagagatc cactttggcg ccggctcgag ggggcccggg 9541 tgcaaagat //