LOCUS Exported 13585 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses catalytically inactive LwCas13a for mammalian RNA targeting. ACCESSION . VERSION . KEYWORDS pC035 - dLwCas13a-msfGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13585) AUTHORS Abudayyeh OO, Gootenberg JS, Essletzbichler P, Han S, Joung J, Belanto JJ, Verdine V, Cox DBT, Kellner MJ, Regev A, Lander ES, Voytas DF, Ting AY, Zhang F TITLE RNA targeting with CRISPR-Cas13. JOURNAL Nature. 2017 Oct 4. doi: 10.1038/nature24049. PUBMED 28976959 REFERENCE 2 (bases 1 to 13585) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13585 /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 2445..3623 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2676..3614 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3571..3591 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 3630..3639 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3645..3665 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4431..7886 /codon_start=1 /product="catalytically dead mutant of a Leptotrichia wadei CRISPR-Cas effector that acts as an RNA-guided RNAse (Abudayyeh et al., 2017)" /label=dLwCas13a /note="contains the inactivating mutations R474A and R1046A" /translation="MKVTKVDGISHKKYIEEGKLVKSTSEENRTSERLSELLSIRLDIY IKNPDNASEEENRIRRENLKKFFSNKVLHLKDSVLYLKNRKEKNAVQDKNYSEEDISEY DLKNKNSFSVLKKILLNEDVNSEELEIFRKDVEAKLNKINSLKYSFEENKANYQKINEN NVEKVGGKSKRNIIYDYYRESAKRNDYINNVQEAFDKLYKKEDIEKLFFLIENSKKHEK YKIREYYHKIIGRKNDKENFAKIIYEEIQNVNNIKELIEKIPDMSELKKSQVFYKYYLD KEELNDKNIKYAFCHFVEIEMSQLLKNYVYKRLSNISNDKIKRIFEYQNLKKLIENKLL NKLDTYVRNCGKYNYYLQVGEIATSDFIARNRQNEAFLRNIIGVSSVAYFSLRNILETE NENGITGRMRGKTVKNNKGEEKYVSGEVDKIYNENKQNEVKENLKMFYSYDFNMDNKNE IEDFFANIDEAISSIAHGIVHFNLELEGKDIFAFKNIAPSEISKKMFQNEINEKKLKLK IFKQLNSANVFNYYEKDVIIKYLKNTKFNFVNKNIPFVPSFTKLYNKIEDLRNTLKFFW SVPKDKEEKDAQIYLLKNIYYGEFLNKFVKNSKVFFKITNEVIKINKQRNQKTGHYKYQ KFENIEKTVPVEYLAIIQSREMINNQDKEEKNTYIDFIQQIFLKGFIDYLNKNNLKYIE SNNNNDNNDIFSKIKIKKDNKEKYDKILKNYEKHNRNKEIPHEINEFVREIKLGKILKY TENLNMFYLILKLLNHKELTNLKGSLEKYQSANKEETFSDELELINLLNLDNNRVTEDF ELEANEIGKFLDFNENKIKDRKELKKFDTNKIYFDGENIIKHRAFYNIKKYGMLNLLEK IADKAKYKISLKELKEYSNKKNEIEKNYTMQQNLHRKYARPKKDEKFNDEDYKEYEKAI GNIQKYTHLKNKVEFNELNLLQGLLLKILHRLVGYTSIWERDLRFRLKGEFPENHYIEE IFNFDNSKNVKYKSGQIVEKYINFYKELYKDNVEKRSIYSDKKVKKLKQEKKDLYIANY IAHFNYIPHAEISLLEVLENLRKLLSYDRKLKNAIMKSIVDILKEYGFVATFKIGADKK IEIQTLESEKIVHLKNLKKKKLMTDRNSEELCELVKVMFEYKALE" CDS 7902..7922 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 7932..8012 /codon_start=1 /product="three tandem HA epitope tags" /label=3xHA /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA" misc_feature 8040..8628 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(8093..8113) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(8511..8522) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(8631..8647) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8632..8648) /label=pBluescriptKS /note="For pBluescript vector" LTR 9153..9333 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(9359..9376) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 9365..9589 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 9635..10063 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(9722..9741) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 9932..9953 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(10072..10092) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 10077..10406 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 10257..10392 /label=SV40 ori /note="SV40 origin of replication" primer_bind 10319..10338 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 10454..10501 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 10520..10894 /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 11024..11145 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(11061..11080) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 11115..11134 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(11194..11210) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(11194..11210) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(11207..11229) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 11218..11234 /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(11242..11272) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 11287..11308 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(11425..11442) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(11596..12184) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(11676..11695) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(12355..13215) /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 12978..12997 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(13216..13320) /gene="bla" /label=AmpR promoter primer_bind complement(13395..13414) /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 ttaatgcaaa gatggataaa gttttaaaca gagaggaatc 2401 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc 2461 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt 2521 gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc 2581 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg 2641 ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg 2701 ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc 2761 agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt 2821 gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc 2881 gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag 2941 ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa 3001 atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg 3061 cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa 3121 tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt 3181 gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa 3241 gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag 3301 agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt 3361 catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt 3421 ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg 3481 agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg 3541 ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt 3601 ttcttccatt tcaggtgtcg tgacgtacgg ccaccatggc tagccccaaa aagaagagga 3661 aagtgggatc cgtgagtaaa ggtgaagaac tcttcactgg agtagtgccc attctggtag 3721 agcttgatgg agatgtaaat ggacataaat tctccgtcag gggcgaaggc gaaggggacg 3781 ccacgaatgg taagctgact ctgaaattca tctgtacgac gggcaaactg cccgtcccat 3841 ggcctacact cgtaacgacc ctcacctacg gcgtgcaatg cttttctcga tatcccgacc 3901 acatgaaaca gcatgacttt ttcaagtctg caatgcctga aggttatgtt caagaaagga 3961 ccatcagctt taaggatgat ggtacatata aaacccgagc cgaggttaaa tttgaagggg 4021 acactctggt taatcgaatt gaactgaaag gtattgattt taaggaggac ggtaacatac 4081 tggggcacaa gttggagtac aactttaaca gccataatgt gtatattacc gctgataagc 4141 agaaaaatgg gataaaggcc aactttaaga tccgacataa tgtcgaagat ggtagtgttc 4201 aactggctga tcattaccaa caaaatacgc ccatcggaga tggacctgta ctcttgcctg 4261 acaatcatta tctctccacg caatcaaagc tttccaagga cccaaacgaa aagagagatc 4321 acatggtcct tctggaattt gtgactgccg caggcatcac tctcggtatg gatgagctgt 4381 acaagggagg aggtggaagc ggaggaggag gaagcggagg aggaggtagc atgaaagtga 4441 ccaaggtcga cggcatcagc cacaagaagt acatcgaaga gggcaagctc gtgaagtcca 4501 ccagcgagga aaaccggacc agcgagagac tgagcgagct gctgagcatc cggctggaca 4561 tctacatcaa gaaccccgac aacgcctccg aggaagagaa ccggatcaga agagagaacc 4621 tgaagaagtt ctttagcaac aaggtgctgc acctgaagga cagcgtgctg tatctgaaga 4681 accggaaaga aaagaacgcc gtgcaggaca agaactatag cgaagaggac atcagcgagt 4741 acgacctgaa aaacaagaac agcttctccg tgctgaagaa gatcctgctg aacgaggacg 4801 tgaactctga ggaactggaa atctttcgga aggacgtgga agccaagctg aacaagatca 4861 acagcctgaa gtacagcttc gaagagaaca aggccaacta ccagaagatc aacgagaaca 4921 acgtggaaaa agtgggcggc aagagcaagc ggaacatcat ctacgactac tacagagaga 4981 gcgccaagcg caacgactac atcaacaacg tgcaggaagc cttcgacaag ctgtataaga 5041 aagaggatat cgagaaactg tttttcctga tcgagaacag caagaagcac gagaagtaca 5101 agatccgcga gtactatcac aagatcatcg gccggaagaa cgacaaagag aacttcgcca 5161 agattatcta cgaagagatc cagaacgtga acaacatcaa agagctgatt gagaagatcc 5221 ccgacatgtc tgagctgaag aaaagccagg tgttctacaa gtactacctg gacaaagagg 5281 aactgaacga caagaatatt aagtacgcct tctgccactt cgtggaaatc gagatgtccc 5341 agctgctgaa aaactacgtg tacaagcggc tgagcaacat cagcaacgat aagatcaagc 5401 ggatcttcga gtaccagaat ctgaaaaagc tgatcgaaaa caaactgctg aacaagctgg 5461 acacctacgt gcggaactgc ggcaagtaca actactatct gcaagtgggc gagatcgcca 5521 cctccgactt tatcgcccgg aaccggcaga acgaggcctt cctgagaaac atcatcggcg 5581 tgtccagcgt ggcctacttc agcctgagga acatcctgga aaccgagaac gagaacggta 5641 tcaccggccg gatgcggggc aagaccgtga agaacaacaa gggcgaagag aaatacgtgt 5701 ccggcgaggt ggacaagatc tacaatgaga acaagcagaa cgaagtgaaa gaaaatctga 5761 agatgttcta cagctacgac ttcaacatgg acaacaagaa cgagatcgag gacttcttcg 5821 ccaatatcga tgaggccatc tctagtattg ctcacggcat cgtgcatttc aacctggaac 5881 tggaaggcaa ggacatcttc gccttcaaga atatcgcccc cagcgagatc tccaagaaga 5941 tgtttcagaa cgaaatcaac gaaaagaagc tgaagctgaa aatcttcaag cagctgaaca 6001 gcgccaacgt gttcaactac tacgagaagg atgtgatcat caagtacctg aagaatacca 6061 agttcaactt cgtgaacaaa aacatcccct tcgtgcccag cttcaccaag ctgtacaaca 6121 agattgagga cctgcggaat accctgaagt ttttttggag cgtgcccaag gacaaagaag 6181 agaaggacgc ccagatctac ctgctgaaga atatctacta cggcgagttc ctgaacaagt 6241 tcgtgaaaaa ctccaaggtg ttctttaaga tcaccaatga agtgatcaag attaacaagc 6301 agcggaacca gaaaaccggc cactacaagt atcagaagtt cgagaacatc gagaaaaccg 6361 tgcccgtgga atacctggcc atcatccaga gcagagagat gatcaacaac caggacaaag 6421 aggaaaagaa tacctacatc gactttattc agcagatttt cctgaagggc ttcatcgact 6481 acctgaacaa gaacaatctg aagtatatcg agagcaacaa caacaatgac aacaacgaca 6541 tcttctccaa gatcaagatc aaaaaggata acaaagagaa gtacgacaag atcctgaaga 6601 actatgagaa gcacaatcgg aacaaagaaa tccctcacga gatcaatgag ttcgtgcgcg 6661 agatcaagct ggggaagatt ctgaagtaca ccgagaatct gaacatgttt tacctgatcc 6721 tgaagctgct gaaccacaaa gagctgacca acctgaaggg cagcctggaa aagtaccagt 6781 ccgccaacaa agaagaaacc ttcagcgacg agttggaact gatcaacctg ctgaacctgg 6841 acaacaacag agtgaccgag gacttcgagc tggaagccaa cgagatcggc aagttcctgg 6901 acttcaacga aaacaaaatc aaggaccgga aagagctgaa aaagttcgac accaacaaga 6961 tctatttcga cggcgagaac atcatcaagc accgggcctt ctacaatatc aagaaatacg 7021 gcatgctgaa tctgctggaa aagatcgccg ataaggccaa gtataagatc agcctgaaag 7081 aactgaaaga gtacagcaac aagaagaatg agattgaaaa gaactacacc atgcagcaga 7141 acctgcaccg gaagtacgcc agacccaaga aggacgaaaa gttcaacgac gaggactaca 7201 aagagtatga gaaggccatc ggcaacatcc agaagtacac ccacctgaag aacaaggtgg 7261 aattcaatga gctgaacctg ctgcagggcc tgctgctgaa gatcctgcac cggctcgtgg 7321 gctacaccag catctgggag cgggacctga gattccggct gaagggcgag tttcccgaga 7381 accactacat cgaggaaatt ttcaatttcg acaactccaa gaatgtgaag tacaaaagcg 7441 gccagatcgt ggaaaagtat atcaacttct acaaagaact gtacaaggac aatgtggaaa 7501 agcggagcat ctactccgac aagaaagtga agaaactgaa gcaggagaag aaagacctgt 7561 atattgcaaa ctacatcgct cactttaatt atattcctca tgccgagatt agcctgctgg 7621 aagtgctgga aaacctgcgg aagctgctgt cctacgaccg gaagctgaag aacgccatca 7681 tgaagtccat cgtggacatt ctgaaagaat acggcttcgt ggccaccttc aagatcggcg 7741 ctgacaagaa gatcgaaatc cagaccctgg aatcagagaa gatcgtgcac ctgaagaatc 7801 tgaagaaaaa gaaactgatg accgaccgga acagcgagga actgtgcgaa ctcgtgaaag 7861 tcatgttcga gtacaaggcc ctggaaggat ccgagggagc tcccaagaaa aagcgcaagg 7921 taggtagttc ctacccatac gatgttccag attacgctta tccctacgac gtgcctgatt 7981 atgcataccc atatgatgtc cccgactatg cctaagaatt cgatatcaag cttatcgata 8041 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 8101 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 8161 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 8221 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 8281 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 8341 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 8401 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg 8461 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 8521 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 8581 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc 8641 gacctcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg 8701 ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc 8761 aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag 8821 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt 8881 ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca gggccaggga 8941 tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag caagagaagg 9001 tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc ctgcatggga 9061 tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta gcatttcatc 9121 acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac cagatctgag 9181 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt 9241 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca 9301 gaccctttta gtcagtgtgg aaaatctcta gcagggcccg tttaaacccg ctgatcagcc 9361 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 9421 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 9481 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 9541 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg 9601 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc 9661 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 9721 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 9781 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 9841 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 9901 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 9961 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 10021 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt 10081 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 10141 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 10201 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 10261 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 10321 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct 10381 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat 10441 ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga 10501 caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg 10561 cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt 10621 ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca 10681 ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct 10741 gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat 10801 gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa 10861 ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac 10921 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat 10981 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc 11041 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 11101 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc 11161 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg 11221 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg 11281 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc 11341 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt 11401 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct 11461 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga 11521 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc 11581 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 11641 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 11701 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 11761 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt 11821 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 11881 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact 11941 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 12001 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct 12061 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 12121 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc 12181 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg 12241 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta 12301 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca 12361 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 12421 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc 12481 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc 12541 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat 12601 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt 12661 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc 12721 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag 12781 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt 12841 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac 12901 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg 12961 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat 13021 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc 13081 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc 13141 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa 13201 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg 13261 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg 13321 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta 13381 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct 13441 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa 13501 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc 13561 gatgtacggg ccagatatac gcgtt //