LOCUS Exported 13550 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral vector encoding Rfx Cas13d fused with 2xNLS, 3xFLAG, and 2A-tagged mCherry.. ACCESSION . VERSION . KEYWORDS pSLQ5428_pHR_EF1a-mCherry-P2A-Rfx_Cas13d-2xNLS-3xFLAG SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13550) AUTHORS Abbott TR, Dhamdhere G, Liu Y, Lin X, Goudy L, Zeng L, Chemparathy A, Chmura S, Heaton NS, Debs R, Pande T, Endy D, La Russa MF, Lewis DB, Qi LS TITLE Development of CRISPR as an Antiviral Strategy to Combat SARS-CoV-2 and Influenza. JOURNAL Cell. 2020 May 14;181(4):865-876.e12. doi: 10.1016/j.cell.2020.04.020. Epub 2020 Apr 29. PUBMED 32353252 REFERENCE 2 (bases 1 to 13550) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13550 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 243..377 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(280..299) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 334..353 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" LTR 545..1178 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 1225..1350 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1847..2080 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2265..2309 /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 2607..2724 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2849..4027 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 3080..4018 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3975..3995 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 4071..4080 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4077..4784 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(4222..4240) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 4490..4509 /label=mCherry-F /note="mCherry, forward primer" CDS 4794..4850 /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 4854..7754 /codon_start=1 /product="RNA-targeting Type VI CRISPR protein from Ruminococcus flavefaciens strain XPD3002 (Konermann et al., 2018)" /label=RfxCas13d /translation="MIEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDS IRSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLE KRYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTV YTYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYII NYGNECYDILALLSGLRHWVVHNNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITN ELTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRK DMSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFV INLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPA GRDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAF FKDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDE NGNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQ KKQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGRE NAEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLE EKGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKK AEEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFRNKAVHLEVARYVH AYINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCV PFGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS" CDS 7764..7784 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 7794..7820 /codon_start=1 /product="nuclear localization signal of human c-Myc proto-oncogene (Dang and Lee, 1988)" /label=c-myc NLS /translation="PAAKRVKLD" CDS 7827..7892 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" misc_feature 7958..8546 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(8011..8031) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(8429..8440) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(8549..8565) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8550..8566) /label=pBluescriptKS /note="For pBluescript vector" LTR 8656..8889 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" promoter complement(8914..8932) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(8915..8932) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(9222..9241) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(9455..9474) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 9574..9596 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(9634..9652) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 9720..9824 /gene="bla" /label=AmpR promoter CDS 9825..10685 /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(10043..10062) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 10856..11444 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 11345..11364 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 11598..11615 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(11685..11705) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 11690..12019 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 11870..12005 /label=SV40 ori /note="SV40 origin of replication" primer_bind 11932..11951 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" intron 13153..13218 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 13348..13368 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" ORIGIN 1 catactgttt tttcttactc cacacaggca tagagtgtct gctattaata actatgctca 61 aaaattgtgt acctttagct ttttaatttg taaaggggtt aataaggaat atttgatgta 121 tagtgccttg actagagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt 181 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg 241 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 301 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 361 cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc aaacttccca 421 ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa acctgtgatt 481 cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact acaaacttag 541 tagttggaag ggctaattca ctcccaaaga agacaagata tccttgatct gtggatctac 601 cacacacaag gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat 661 ccactgacct ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag 721 gccaataaag gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac 781 ccggagagag aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc 841 cgagagctgc atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt 901 tccgctgggg actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc 961 agatcctgca tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat 1021 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 1081 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 1141 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg 1201 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc 1261 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc 1321 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga 1381 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta 1441 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa 1501 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt 1561 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa 1621 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca 1681 cagcaagcgg ccggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga 1741 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca 1801 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc 1861 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac 1921 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg 1981 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa 2041 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg 2101 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg 2161 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa 2221 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat 2281 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt 2341 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg 2401 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc 2461 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga 2521 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc 2581 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt 2641 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa 2701 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg 2761 ttaattaatg caaagatgga taaagtttta aacagagagg aatctttgca gctaatggac 2821 cttctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac 2881 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag 2941 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga 3001 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg 3061 gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac 3121 gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg tgattcttga 3181 tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt aaggagcccc 3241 ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg 3301 gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt aaaatttttg 3361 atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg gccaagatct 3421 gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg 3481 cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac gggggtagtc 3541 tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg 3601 ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg 3661 ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg cgggtgagtc 3721 acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg actccacgga 3781 gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta cgtcgtcttt 3841 aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg tggagactga 3901 agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg 3961 atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc catttcaggt 4021 gtcgtgaacg cgtctgcagg atacaagctt gcggtaccgc gggcgcgccc gccaccatgg 4081 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc 4141 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc 4201 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg 4261 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg 4321 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga 4381 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg 4441 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc 4501 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc 4561 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg 4621 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca 4681 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg 4741 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggcagc ggagctacta 4801 acttcagcct gctgaagcag gctggagacg tggaggagaa ccctggaccc gggatgatcg 4861 agaaaaagaa gtccttcgct aagggcatgg gcgtgaagtc cacactcgtg tccggctcca 4921 aagtgtacat gacaaccttc gccgaaggca gcgacgccag gctggaaaag atcgtggagg 4981 gcgacagcat caggagcgtg aatgagggcg aggccttcag cgctgaaatg gccgataaaa 5041 acgccggcta taagatcggc aacgccaaat tcagccatcc taagggctac gccgtggtgg 5101 ctaacaaccc tctgtataca ggacccgtcc agcaggatat gctcggcctg aaggaaactc 5161 tggaaaagag gtacttcggc gagagcgctg atggcaatga caatatttgt atccaggtga 5221 tccataacat cctggacatt gaaaaaatcc tcgccgaata cattaccaac gccgcctacg 5281 ccgtcaacaa tatctccggc ctggataagg acattattgg attcggcaag ttctccacag 5341 tgtataccta cgacgaattc aaagaccccg agcaccatag ggccgctttc aacaataacg 5401 ataagctcat caacgccatc aaggcccagt atgacgagtt cgacaacttc ctcgataacc 5461 ccagactcgg ctatttcggc caggcctttt tcagcaagga gggcagaaat tacatcatca 5521 attacggcaa cgaatgctat gacattctgg ccctcctgag cggactgaga cactgggtgg 5581 tccacaacaa cgaagaagag tccaggatct ccaggacctg gctctacaac ctcgataaga 5641 acctcgacaa cgaatacatc tccaccctca actacctcta cgacaggatc accaatgagc 5701 tgaccaactc cttctccaag aactccgccg ccaacgtgaa ctatattgcc gaaactctgg 5761 gaatcaaccc tgccgaattc gccgaacaat atttcagatt cagcattatg aaagagcaga 5821 aaaacctcgg attcaatatc accaagctca gggaagtgat gctggacagg aaggatatgt 5881 ccgagatcag gaaaaatcat aaggtgttcg actccatcag gaccaaggtc tacaccatga 5941 tggactttgt gatttatagg tattacatcg aagaggatgc caaggtggct gccgccaata 6001 agtccctccc cgataatgag aagtccctga gcgagaagga tatctttgtg attaacctga 6061 ggggctcctt caacgacgac cagaaggatg ccctctacta cgatgaagct aatagaattt 6121 ggagaaagct cgaaaatatc atgcacaaca tcaaggaatt taggggaaac aagacaagag 6181 agtataagaa gaaggacgcc cctagactgc ccagaatcct gcccgctggc cgtgatgttt 6241 ccgccttcag caaactcatg tatgccctga caatgttcct ggatggcaag gagatcaacg 6301 acctcctgac caccctgatt aataaattcg ataacatcca gagcttcctg aaggtgatgc 6361 ctctcatcgg agtcaacgct aagttcgtgg aggaatacgc ctttttcaaa gactccgcca 6421 agatcgccga tgagctgagg ctgatcaagt ccttcgctag aatgggagaa cctattgccg 6481 atgccaggag ggccatgtat atcgacgcca tccgtatttt aggaaccaac ctgtcctatg 6541 atgagctcaa ggccctcgcc gacacctttt ccctggacga gaacggaaac aagctcaaga 6601 aaggcaagca cggcatgaga aatttcatta ttaataacgt gatcagcaat aaaaggttcc 6661 actacctgat cagatacggt gatcctgccc acctccatga gatcgccaaa aacgaggccg 6721 tggtgaagtt cgtgctcggc aggatcgctg acatccagaa aaaacagggc cagaacggca 6781 agaaccagat cgacaggtac tacgaaactt gtatcggaaa ggataagggc aagagcgtga 6841 gcgaaaaggt ggacgctctc acaaagatca tcaccggaat gaactacgac caattcgaca 6901 agaaaaggag cgtcattgag gacaccggca gggaaaacgc cgagagggag aagtttaaaa 6961 agatcatcag cctgtacctc accgtgatct accacatcct caagaatatt gtcaatatca 7021 acgccaggta cgtcatcgga ttccattgcg tcgagcgtga tgctcaactg tacaaggaga 7081 aaggctacga catcaatctc aagaaactgg aagagaaggg attcagctcc gtcaccaagc 7141 tctgcgctgg cattgatgaa actgcccccg ataagagaaa ggacgtggaa aaggagatgg 7201 ctgaaagagc caaggagagc attgacagcc tcgagagcgc caaccccaag ctgtatgcca 7261 attacatcaa atacagcgac gagaagaaag ccgaggagtt caccaggcag attaacaggg 7321 agaaggccaa aaccgccctg aacgcctacc tgaggaacac caagtggaat gtgatcatca 7381 gggaggacct cctgagaatt gacaacaaga catgtaccct gttcaggaac aaggccgtcc 7441 atctggaagt ggccaggtat gtccacgcct atatcaacga cattgccgag gtcaattcct 7501 acttccaact gtaccattac atcatgcaga gaattatcat gaatgagagg tacgagaaaa 7561 gcagcggaaa ggtgtccgag tacttcgacg ctgtgaatga cgagaagaag tacaacgata 7621 ggctcctgaa actgctgtgt gtgcctttcg gctactgtat ccccaggttt aagaacctga 7681 gcatcgaggc cctgttcgat aggaacgagg ccgccaagtt cgacaaggag aaaaagaaag 7741 tgtccggcaa ttccactagt gctcccaaga aaaagcgcaa ggtaggtgga agcccagcag 7801 ctaaaagagt taaattggat ggatccgatt ataaggatca cgatggagat tacaaggacc 7861 acgacataga ctacaaagat gatgacgaca agtaacgtac ggccgcgact ctagagtcga 7921 cctgcaggca tgcaagcttg atatcaagct tatcgataat caacctctgg attacaaaat 7981 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc 8041 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt 8101 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg 8161 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg 8221 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc 8281 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt 8341 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct 8401 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg 8461 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg 8521 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgaggga attaattcga 8581 gctcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa 8641 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg 8701 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag 8761 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc 8821 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa 8881 tctctagcag catctagaat taattccgtg tattctatag tgtcacctaa atcgtatgtg 8941 tatgatacat aaggttatgt attaattgta gccgcgttct aacgacaata tgtacaagcc 9001 taattgtgta gcatctggct tactgaagca gaccctatca tctctctcgt aaactgccgt 9061 cagagtcggt ttggttggac gaaccttctg agtttctggt aacgccgtcc cgcacccgga 9121 aatggtcagc gaaccaatca gcagggtcat cgctagccag atcctctacg ccggacgcat 9181 cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac 9241 cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat 9301 ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct 9361 tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc 9421 gcataaggga gagcgtcgaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt 9481 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc 9541 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt 9601 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg 9661 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc 9721 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac 9781 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt 9841 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag 9901 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg 9961 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa 10021 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc 10081 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag 10141 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa 10201 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc 10261 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg 10321 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa 10381 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa 10441 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg 10501 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag 10561 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg 10621 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt 10681 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt 10741 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac 10801 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 10861 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 10921 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 10981 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga 11041 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 11101 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 11161 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 11221 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 11281 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 11341 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 11401 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 11461 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat 11521 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca 11581 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca 11641 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctgtggaatg tgtgtcagtt 11701 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 11761 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 11821 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct 11881 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 11941 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg 12001 aggcctaggc ttttgcaaaa agcttggaca caagacaggc ttgcgagata tgtttgagaa 12061 taccacttta tcccgcgtca gggagaggca gtgcgtaaaa agacgcggac tcatgtgaaa 12121 tactggtttt tagtgcgcca gatctctata atctcgcgca acctattttc ccctcgaaca 12181 ctttttaagc cgtagataaa caggctggga cacttcacat gagcgaaaaa tacatcgtca 12241 cctgggacat gttgcagatc catgcacgta aactcgcaag ccgactgatg ccttctgaac 12301 aatggaaagg cattattgcc gtaagccgtg gcggtctgta ccgggtgcgt tactggcgcg 12361 tgaactgggt attcgtcatg tcgataccgt ttgtatttcc agctacgatc acgacaacca 12421 gcgcgagctt aaagtgctga aacgcgcaga aggcgatggc gaaggcttca tcgttattga 12481 tgacctggtg gataccggtg gtactgcggt tgcgattcgt gaaatgtatc caaaagcgca 12541 ctttgtcacc atcttcgcaa aaccggctgg tcgtccgctg gttgatgact atgttgttga 12601 tatcccgcaa gatacctgga ttgaacagcc gtgggatatg ggcgtcgtat tcgtcccgcc 12661 aatctccggt cgctaatctt ttcaacgcct ggcactgccg ggcgttgttc tttttaactt 12721 caggcgggtt acaatagttt ccagtaagta ttctggaggc tgcatccatg acacaggcaa 12781 acctgagcga aaccctgttc aaaccccgct ttaaacatcc tgaaacctcg acgctagtcc 12841 gccgctttaa tcacggcgca caaccgcctg tgcagtcggc ccttgatggt aaaaccatcc 12901 ctcactggta tcgcatgatt aaccgtctga tgtggatctg gcgcggcatt gacccacgcg 12961 aaatcctcga cgtccaggca cgtattgtga tgagcgatgc cgaacgtacc gacgatgatt 13021 tatacgatac ggtgattggc taccgtggcg gcaactggat ttatgagtgg gccccggatc 13081 tttgtgaagg aaccttactt ctgtggtgtg acataattgg acaaactacc tacagagatt 13141 taaagctcta aggtaaatat aaaattttta agtgtataat gtgttaaact actgattcta 13201 attgtttgtg tattttagat tccaacctat ggaactgatg aatgggagca gtggtggaat 13261 gcctttaatg aggaaaacct gttttgctca gaagaaatgc catctagtga tgatgaggct 13321 actgctgact ctcaacattc tactcctcca aaaaagaaga gaaaggtaga agaccccaag 13381 gactttcctt cagaattgct aagttttttg agtcatgctg tgtttagtaa tagaactctt 13441 gcttgctttg ctatttacac cacaaaggaa aaagctgcac tgctatacaa gaaaattatg 13501 gaaaaatatt ctgtaacctt tataagtagg cataacagtt ataatcataa //