LOCUS Exported 12863 bp ds-DNA circular SYN 01-JUN-2022 DEFINITION Lenti virus delivery of AsCpf1 and crRNA guide. ACCESSION . VERSION . KEYWORDS pY108 (lenti-AsCpf1) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12863) AUTHORS Zetsche B, Heidenreich M, Mohanraju P, Fedorova I, Kneppers J, DeGennaro EM, Winblad N, Choudhury SR, Abudayyeh OO, Gootenberg JS, Wu WY, Scott DA, Severinov K, van der Oost J, Zhang F TITLE Multiplex gene editing by CRISPR-Cpf1 using a single crRNA array. JOURNAL Nat Biotechnol. 2017 Jan;35(1):31-34. doi: 10.1038/nbt.3737. Epub 2016 Dec 5. PUBMED 27918548 REFERENCE 2 (bases 1 to 12863) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 1, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12863 /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 complement(2422..2662) /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind complement(2472..2491) /label=LKO.1 5' /note="Human U6 promoter, forward primer" primer_bind complement(2642..2662) /label=hU6-F /note="Human U6 promoter, forward primer" promoter 2711..2922 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" regulatory 2930..2939 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2942..2962 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2987..6904 /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 6624..6633 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 6905..6952 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 6959..7039 /codon_start=1 /product="three tandem HA epitope tags" /label=3xHA /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA" CDS 7049..7105 /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 7106..7702 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind 7599..7619 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 7718..8306 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(7771..7791) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(8189..8200) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 8378..8611 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(8637..8654) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 8643..8867 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 8913..9341 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(9000..9019) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 9210..9231 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(9350..9370) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 9355..9684 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 9535..9670 /label=SV40 ori /note="SV40 origin of replication" primer_bind 9597..9616 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 9732..9779 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 9798..10172 /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 10302..10423 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(10339..10358) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 10393..10412 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(10472..10488) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(10472..10488) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(10485..10507) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 10496..10512 /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(10520..10550) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 10565..10586 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(10703..10720) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(10874..11462) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(10954..10973) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(11633..12493) /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 12256..12275 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(12494..12598) /gene="bla" /label=AmpR promoter primer_bind complement(12673..12692) /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 tttaattaaa aaagagacga tatatcgtct ccatctacaa 2401 gagtagaaat tacggtgttt cgtcctttcc acaagatata taaagccaag aaatcgaaat 2461 actttcaagt tacggtaagc atatgatagt ccattttaaa acataatttt aaaactgcaa 2521 actacccaag aaattattac tttctacgtc acgtattttg tactaatatc tttgtgttta 2581 cagtcaaatt aattccaatt atctctctaa cagccttgta tcgtatatgc aaatatgaag 2641 gaatcatggg aaataggccc tcctcgagag gtcttgaaag gagtgggaat tggctccggt 2701 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc 2761 ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg 2821 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc 2881 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggaccggtg ccaccatggc 2941 cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccacac agttcgaggg 3001 ctttaccaac ctgtatcagg tgagcaagac actgcggttt gagctgatcc cacagggcaa 3061 gaccctgaag cacatccagg agcagggctt catcgaggag gacaaggccc gcaatgatca 3121 ctacaaggag ctgaagccca tcatcgatcg gatctacaag acctatgccg accagtgcct 3181 gcagctggtg cagctggatt gggagaacct gagcgccgcc atcgactcct atagaaagga 3241 gaaaaccgag gagacaagga acgccctgat cgaggagcag gccacatatc gcaatgccat 3301 ccacgactac ttcatcggcc ggacagacaa cctgaccgat gccatcaata agagacacgc 3361 cgagatctac aagggcctgt tcaaggccga gctgtttaat ggcaaggtgc tgaagcagct 3421 gggcaccgtg accacaaccg agcacgagaa cgccctgctg cggagcttcg acaagtttac 3481 aacctacttc tccggctttt atgagaacag gaagaacgtg ttcagcgccg aggatatcag 3541 cacagccatc ccacaccgca tcgtgcagga caacttcccc aagtttaagg agaattgtca 3601 catcttcaca cgcctgatca ccgccgtgcc cagcctgcgg gagcactttg agaacgtgaa 3661 gaaggccatc ggcatcttcg tgagcacctc catcgaggag gtgttttcct tcccttttta 3721 taaccagctg ctgacacaga cccagatcga cctgtataac cagctgctgg gaggaatctc 3781 tcgggaggca ggcaccgaga agatcaaggg cctgaacgag gtgctgaatc tggccatcca 3841 gaagaatgat gagacagccc acatcatcgc ctccctgcca cacagattca tccccctgtt 3901 taagcagatc ctgtccgata ggaacaccct gtctttcatc ctggaggagt ttaagagcga 3961 cgaggaagtg atccagtcct tctgcaagta caagacactg ctgagaaacg agaacgtgct 4021 ggagacagcc gaggccctgt ttaacgagct gaacagcatc gacctgacac acatcttcat 4081 cagccacaag aagctggaga caatcagcag cgccctgtgc gaccactggg atacactgag 4141 gaatgccctg tatgagcgga gaatctccga gctgacaggc aagatcacca agtctgccaa 4201 ggagaaggtg cagcgcagcc tgaagcacga ggatatcaac ctgcaggaga tcatctctgc 4261 cgcaggcaag gagctgagcg aggccttcaa gcagaaaacc agcgagatcc tgtcccacgc 4321 acacgccgcc ctggatcagc cactgcctac aaccctgaag aagcaggagg agaaggagat 4381 cctgaagtct cagctggaca gcctgctggg cctgtaccac ctgctggact ggtttgccgt 4441 ggatgagtcc aacgaggtgg accccgagtt ctctgcccgg ctgaccggca tcaagctgga 4501 gatggagcct tctctgagct tctacaacaa ggccagaaat tatgccacca agaagcccta 4561 ctccgtggag aagttcaagc tgaactttca gatgcctaca ctggcctctg gctgggacgt 4621 gaataaggag aagaacaatg gcgccatcct gtttgtgaag aacggcctgt actatctggg 4681 catcatgcca aagcagaagg gcaggtataa ggccctgagc ttcgagccca cagagaaaac 4741 cagcgagggc tttgataaga tgtactatga ctacttccct gatgccgcca agatgatccc 4801 aaagtgcagc acccagctga aggccgtgac agcccacttt cagacccaca caacccccat 4861 cctgctgtcc aacaatttca tcgagcctct ggagatcaca aaggagatct acgacctgaa 4921 caatcctgag aaggagccaa agaagtttca gacagcctac gccaagaaaa ccggcgacca 4981 gaagggctac agagaggccc tgtgcaagtg gatcgacttc acaagggatt ttctgtccaa 5041 gtataccaag acaacctcta tcgatctgtc tagcctgcgg ccatcctctc agtataagga 5101 cctgggcgag tactatgccg agctgaatcc cctgctgtac cacatcagct tccagagaat 5161 cgccgagaag gagatcatgg atgccgtgga gacaggcaag ctgtacctgt tccagatcta 5221 taacaaggac tttgccaagg gccaccacgg caagcctaat ctgcacacac tgtattggac 5281 cggcctgttt tctccagaga acctggccaa gacaagcatc aagctgaatg gccaggccga 5341 gctgttctac cgccctaagt ccaggatgaa gaggatggca caccggctgg gagagaagat 5401 gctgaacaag aagctgaagg atcagaaaac cccaatcccc gacaccctgt accaggagct 5461 gtacgactat gtgaatcaca gactgtccca cgacctgtct gatgaggcca gggccctgct 5521 gcccaacgtg atcaccaagg aggtgtctca cgagatcatc aaggataggc gctttaccag 5581 cgacaagttc tttttccacg tgcctatcac actgaactat caggccgcca attccccatc 5641 taagttcaac cagagggtga atgcctacct gaaggagcac cccgagacac ctatcatcgg 5701 catcgatcgg ggcgagagaa acctgatcta tatcacagtg atcgactcca ccggcaagat 5761 cctggagcag cggagcctga acaccatcca gcagtttgat taccagaaga agctggacaa 5821 cagggagaag gagagggtgg cagcaaggca ggcctggtct gtggtgggca caatcaagga 5881 tctgaagcag ggctatctga gccaggtcat ccacgagatc gtggacctga tgatccacta 5941 ccaggccgtg gtggtgctgg agaacctgaa tttcggcttt aagagcaaga ggaccggcat 6001 cgccgagaag gccgtgtacc agcagttcga gaagatgctg atcgataagc tgaattgcct 6061 ggtgctgaag gactatccag cagagaaagt gggaggcgtg ctgaacccat accagctgac 6121 agaccagttc acctcctttg ccaagatggg cacccagtct ggcttcctgt tttacgtgcc 6181 tgccccatat acatctaaga tcgatcccct gaccggcttc gtggacccct tcgtgtggaa 6241 aaccatcaag aatcacgaga gccgcaagca cttcctggag ggcttcgact ttctgcacta 6301 cgacgtgaaa accggcgact tcatcctgca ctttaagatg aacagaaatc tgtccttcca 6361 gaggggcctg cccggcttta tgcctgcatg ggatatcgtg ttcgagaaga acgagacaca 6421 gtttgacgcc aagggcaccc ctttcatcgc cggcaagaga atcgtgccag tgatcgagaa 6481 tcacagattc accggcagat accgggacct gtatcctgcc aacgagctga tcgccctgct 6541 ggaggagaag ggcatcgtgt tcagggatgg ctccaacatc ctgccaaagc tgctggagaa 6601 tgacgattct cacgccatcg acaccatggt ggccctgatc cgcagcgtgc tgcagatgcg 6661 gaactccaat gccgccacag gcgaggacta tatcaacagc cccgtgcgcg atctgaatgg 6721 cgtgtgcttc gactcccggt ttcagaaccc agagtggccc atggacgccg atgccaatgg 6781 cgcctaccac atcgccctga agggccagct gctgctgaat cacctgaagg agagcaagga 6841 tctgaagctg cagaacggca tctccaatca ggactggctg gcctacatcc aggagctgcg 6901 caacaaaagg ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa agggatccta 6961 cccatacgat gttccagatt acgcttatcc ctacgacgtg cctgattatg catacccata 7021 tgatgtcccc gactatgccg gatccggcgc aacaaacttc tctctgctga aacaagccgg 7081 agatgtcgaa gagaatcctg gaccgaccga gtacaagccc acggtgcgcc tcgccacccg 7141 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac 7201 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt 7261 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt 7321 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc 7381 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct 7441 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggag tctcgcccga 7501 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg 7561 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg 7621 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat 7681 gacccgcaag cccggtgcct gaacgcgtta agtcgacaat caacctctgg attacaaaat 7741 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc 7801 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt 7861 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg 7921 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg 7981 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc 8041 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt 8101 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct 8161 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg 8221 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg 8281 gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct 8341 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa 8401 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc 8461 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga 8521 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga 8581 cccttttagt cagtgtggaa aatctctagc agggcccgtt taaacccgct gatcagcctc 8641 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac 8701 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg 8761 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga 8821 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga 8881 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc 8941 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc 9001 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 9061 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa 9121 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 9181 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact 9241 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg 9301 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt 9361 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 9421 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg 9481 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg 9541 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt 9601 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt 9661 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct 9721 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca 9781 aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg 9841 acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg 9901 aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg 9961 accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt 10021 acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga 10081 ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact 10141 gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg 10201 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 10261 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 10321 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 10381 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 10441 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 10501 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 10561 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 10621 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 10681 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 10741 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 10801 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 10861 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 10921 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 10981 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 11041 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 11101 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 11161 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 11221 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 11281 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 11341 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 11401 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 11461 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 11521 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 11581 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 11641 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 11701 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 11761 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 11821 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 11881 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 11941 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 12001 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 12061 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 12121 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 12181 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 12241 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 12301 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 12361 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 12421 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 12481 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 12541 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 12601 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg 12661 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc 12721 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg 12781 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga 12841 tgtacgggcc agatatacgc gtt //