LOCUS Exported 12656 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral plasmid for expressing human codon-optimized AsCpf1 containing 3x N-terminal MYC-NLS signal. ACCESSION . VERSION . KEYWORDS LentiAsCpf1-3xMYC-blast SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12656) AUTHORS Liu J, Srinivasan S, Li CY, Ho IL, Rose J, Shaheen M, Wang G, Yao W, Deem A, Bristow C, Hart T, Draetta G TITLE Pooled library screening with multiplexed Cpf1 library. JOURNAL Nat Commun. 2019 Jul 17;10(1):3144. doi: 10.1038/s41467-019-10963-x. PUBMED 31316073 REFERENCE 2 (bases 1 to 12656) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12656 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 382..580 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 598..778 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 825..950 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1443..1676 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1861..1905 /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 2203..2320 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2413..2624 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" regulatory 2632..2641 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2644..2670 /codon_start=1 /product="nuclear localization signal of human c-Myc proto-oncogene (Dang and Lee, 1988)" /label=c-myc NLS /translation="PAAKRVKLD" CDS 2671..2697 /codon_start=1 /product="nuclear localization signal of human c-Myc proto-oncogene (Dang and Lee, 1988)" /label=c-myc NLS /translation="PAAKRVKLD" CDS 2698..2724 /codon_start=1 /product="nuclear localization signal of human c-Myc proto-oncogene (Dang and Lee, 1988)" /label=c-myc NLS /translation="PAAKRVKLD" CDS 2725..6642 /codon_start=1 /product="CRISPR-associated protein from Acidaminococcus sp. BV3L6 (Zetsche et al., 2015)" /label=AsCpf1 /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK FTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR NYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKA LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK DLKLQNGISNQDWLAYIQELRN" regulatory 6362..6371 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 6652..6708 /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 6709..7107 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" misc_feature 7132..7720 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(7185..7205) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(7603..7614) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(7723..7739) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7724..7740) /label=pBluescriptKS /note="For pBluescript vector" LTR 8245..8425 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(8451..8468) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 8457..8681 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 8727..9155 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8814..8833) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 9024..9045 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(9164..9184) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 9329..9463 /label=SV40 ori /note="SV40 origin of replication" primer_bind 9390..9409 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 9525..9572 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 9591..9965 /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 10095..10216 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(10132..10151) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 10186..10205 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(10265..10281) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(10265..10281) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(10278..10300) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 10289..10305 /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(10313..10343) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 10358..10379 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(10496..10513) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(10667..11255) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(10747..10766) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(11426..12286) /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 12049..12068 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(12287..12391) /gene="bla" /label=AmpR promoter primer_bind complement(12466..12485) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg 601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga 841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa 901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc 961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat 1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg 1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc 1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc 1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa 1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat 1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta 1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga 1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca 1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat 1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag 1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg 1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg 1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa 1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa 1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac 1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt 1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca 2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa 2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg 2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg 2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac 2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag 2341 cagagatcca gtttggttaa ttagctagct aggtcttgaa aggagtggga attggctccg 2401 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg 2461 tcggcaattg atccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg 2521 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg 2581 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggaccgg tgccaccatg 2641 gcgccagcgg caaagcgcgt aaaattggac ccagcagcca aacgagtaaa actcgatccg 2701 gctgctaaac gagttaagct ggacacacag ttcgagggct ttaccaacct gtatcaggtg 2761 agcaagacac tgcggtttga gctgatccca cagggcaaga ccctgaagca catccaggag 2821 cagggcttca tcgaggagga caaggcccgc aatgatcact acaaggagct gaagcccatc 2881 atcgatcgga tctacaagac ctatgccgac cagtgcctgc agctggtgca gctggattgg 2941 gagaacctga gcgccgccat cgactcctat agaaaggaga aaaccgagga gacaaggaac 3001 gccctgatcg aggagcaggc cacatatcgc aatgccatcc acgactactt catcggccgg 3061 acagacaacc tgaccgatgc catcaataag agacacgccg agatctacaa gggcctgttc 3121 aaggccgagc tgtttaatgg caaggtgctg aagcagctgg gcaccgtgac cacaaccgag 3181 cacgagaacg ccctgctgcg gagcttcgac aagtttacaa cctacttctc cggcttttat 3241 gagaacagga agaacgtgtt cagcgccgag gatatcagca cagccatccc acaccgcatc 3301 gtgcaggaca acttccccaa gtttaaggag aattgtcaca tcttcacacg cctgatcacc 3361 gccgtgccca gcctgcggga gcactttgag aacgtgaaga aggccatcgg catcttcgtg 3421 agcacctcca tcgaggaggt gttttccttc cctttttata accagctgct gacacagacc 3481 cagatcgacc tgtataacca gctgctggga ggaatctctc gggaggcagg caccgagaag 3541 atcaagggcc tgaacgaggt gctgaatctg gccatccaga agaatgatga gacagcccac 3601 atcatcgcct ccctgccaca cagattcatc cccctgttta agcagatcct gtccgatagg 3661 aacaccctgt ctttcatcct ggaggagttt aagagcgacg aggaagtgat ccagtccttc 3721 tgcaagtaca agacactgct gagaaacgag aacgtgctgg agacagccga ggccctgttt 3781 aacgagctga acagcatcga cctgacacac atcttcatca gccacaagaa gctggagaca 3841 atcagcagcg ccctgtgcga ccactgggat acactgagga atgccctgta tgagcggaga 3901 atctccgagc tgacaggcaa gatcaccaag tctgccaagg agaaggtgca gcgcagcctg 3961 aagcacgagg atatcaacct gcaggagatc atctctgccg caggcaagga gctgagcgag 4021 gccttcaagc agaaaaccag cgagatcctg tcccacgcac acgccgccct ggatcagcca 4081 ctgcctacaa ccctgaagaa gcaggaggag aaggagatcc tgaagtctca gctggacagc 4141 ctgctgggcc tgtaccacct gctggactgg tttgccgtgg atgagtccaa cgaggtggac 4201 cccgagttct ctgcccggct gaccggcatc aagctggaga tggagccttc tctgagcttc 4261 tacaacaagg ccagaaatta tgccaccaag aagccctact ccgtggagaa gttcaagctg 4321 aactttcaga tgcctacact ggcctctggc tgggacgtga ataaggagaa gaacaatggc 4381 gccatcctgt ttgtgaagaa cggcctgtac tatctgggca tcatgccaaa gcagaagggc 4441 aggtataagg ccctgagctt cgagcccaca gagaaaacca gcgagggctt tgataagatg 4501 tactatgact acttccctga tgccgccaag atgatcccaa agtgcagcac ccagctgaag 4561 gccgtgacag cccactttca gacccacaca acccccatcc tgctgtccaa caatttcatc 4621 gagcctctgg agatcacaaa ggagatctac gacctgaaca atcctgagaa ggagccaaag 4681 aagtttcaga cagcctacgc caagaaaacc ggcgaccaga agggctacag agaggccctg 4741 tgcaagtgga tcgacttcac aagggatttt ctgtccaagt ataccaagac aacctctatc 4801 gatctgtcta gcctgcggcc atcctctcag tataaggacc tgggcgagta ctatgccgag 4861 ctgaatcccc tgctgtacca catcagcttc cagagaatcg ccgagaagga gatcatggat 4921 gccgtggaga caggcaagct gtacctgttc cagatctata acaaggactt tgccaagggc 4981 caccacggca agcctaatct gcacacactg tattggaccg gcctgttttc tccagagaac 5041 ctggccaaga caagcatcaa gctgaatggc caggccgagc tgttctaccg ccctaagtcc 5101 aggatgaaga ggatggcaca ccggctggga gagaagatgc tgaacaagaa gctgaaggat 5161 cagaaaaccc caatccccga caccctgtac caggagctgt acgactatgt gaatcacaga 5221 ctgtcccacg acctgtctga tgaggccagg gccctgctgc ccaacgtgat caccaaggag 5281 gtgtctcacg agatcatcaa ggataggcgc tttaccagcg acaagttctt tttccacgtg 5341 cctatcacac tgaactatca ggccgccaat tccccatcta agttcaacca gagggtgaat 5401 gcctacctga aggagcaccc cgagacacct atcatcggca tcgatcgggg cgagagaaac 5461 ctgatctata tcacagtgat cgactccacc ggcaagatcc tggagcagcg gagcctgaac 5521 accatccagc agtttgatta ccagaagaag ctggacaaca gggagaagga gagggtggca 5581 gcaaggcagg cctggtctgt ggtgggcaca atcaaggatc tgaagcaggg ctatctgagc 5641 caggtcatcc acgagatcgt ggacctgatg atccactacc aggccgtggt ggtgctggag 5701 aacctgaatt tcggctttaa gagcaagagg accggcatcg ccgagaaggc cgtgtaccag 5761 cagttcgaga agatgctgat cgataagctg aattgcctgg tgctgaagga ctatccagca 5821 gagaaagtgg gaggcgtgct gaacccatac cagctgacag accagttcac ctcctttgcc 5881 aagatgggca cccagtctgg cttcctgttt tacgtgcctg ccccatatac atctaagatc 5941 gatcccctga ccggcttcgt ggaccccttc gtgtggaaaa ccatcaagaa tcacgagagc 6001 cgcaagcact tcctggaggg cttcgacttt ctgcactacg acgtgaaaac cggcgacttc 6061 atcctgcact ttaagatgaa cagaaatctg tccttccaga ggggcctgcc cggctttatg 6121 cctgcatggg atatcgtgtt cgagaagaac gagacacagt ttgacgccaa gggcacccct 6181 ttcatcgccg gcaagagaat cgtgccagtg atcgagaatc acagattcac cggcagatac 6241 cgggacctgt atcctgccaa cgagctgatc gccctgctgg aggagaaggg catcgtgttc 6301 agggatggct ccaacatcct gccaaagctg ctggagaatg acgattctca cgccatcgac 6361 accatggtgg ccctgatccg cagcgtgctg cagatgcgga actccaatgc cgccacaggc 6421 gaggactata tcaacagccc cgtgcgcgat ctgaatggcg tgtgcttcga ctcccggttt 6481 cagaacccag agtggcccat ggacgccgat gccaatggcg cctaccacat cgccctgaag 6541 ggccagctgc tgctgaatca cctgaaggag agcaaggatc tgaagctgca gaacggcatc 6601 tccaatcagg actggctggc ctacatccag gagctgcgca acggatccgg cgcaacaaac 6661 ttctctctgc tgaaacaagc cggagatgtc gaagagaatc ctggaccgat ggccaagcct 6721 ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc 6781 atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact 6841 ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact 6901 gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg 6961 ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc 7021 aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg 7081 ccctctggtt atgtgtggga gggctaagaa ttcgatatca agcttatcgg taatcaacct 7141 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 7201 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 7261 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 7321 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 7381 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 7441 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 7501 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 7561 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 7621 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 7681 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga 7741 gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt 7801 gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct 7861 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg 7921 ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac 7981 cacacacaag gctacttccc tgattggcag aactacacac cagggccagg gatcagatat 8041 ccactgacct ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa 8101 gccaatgaag gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac 8161 ccggagagag aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc 8221 cgagagctgc atccggactg tactgggtct ctctggttag accagatctg agcctgggag 8281 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt 8341 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt 8401 tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag cctcgactgt 8461 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 8521 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 8581 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 8641 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac 8701 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg 8761 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 8821 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 8881 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 8941 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 9001 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 9061 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 9121 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta 9181 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 9241 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 9301 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccag ttccgcccat 9361 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 9421 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 9481 ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat 9541 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt 9601 tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga 9661 ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg 9721 acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg 9781 cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca 9841 cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc 9901 gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg 9961 actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg 10021 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt 10081 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 10141 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 10201 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat 10261 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 10321 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 10381 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 10441 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 10501 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 10561 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 10621 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 10681 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 10741 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 10801 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 10861 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 10921 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 10981 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 11041 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 11101 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 11161 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 11221 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 11281 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 11341 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 11401 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 11461 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 11521 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 11581 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 11641 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 11701 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 11761 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 11821 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 11881 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 11941 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 12001 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 12061 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 12121 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 12181 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 12241 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 12301 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 12361 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 12421 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 12481 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 12541 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 12601 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtt //