LOCUS Exported 13584 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 13584) 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 13584) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13584 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(47..66) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 238..617 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 619..817 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 772..792 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 835..1015 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1062..1187 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1680..1913 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2098..2142 /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 2440..2557 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2682..3859 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2913..3850 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3807..3827 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 3866..3875 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3881..3901 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4667..8122 /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 8138..8158 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 8168..8248 /codon_start=1 /product="three tandem HA epitope tags" /label=3xHA /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA" misc_feature 8276..8864 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(8329..8349) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(8747..8758) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(8867..8883) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8868..8884) /label=pBluescriptKS /note="For pBluescript vector" LTR 9389..9569 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(9595..9612) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 9601..9825 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 9871..10299 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(9958..9977) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 10168..10189 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(10308..10328) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 10313..10642 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 10493..10628 /label=SV40 ori /note="SV40 origin of replication" primer_bind 10555..10574 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 10690..10737 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 10756..11130 /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 11260..11381 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(11297..11316) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 11351..11370 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(11430..11446) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(11430..11446) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(11443..11465) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 11454..11470 /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(11478..11508) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 11523..11544 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(11661..11678) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(11832..12420) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(11912..11931) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(12591..13451) /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 13214..13233 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(13452..13556) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg 61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac 241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct 841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg 1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga 1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata 1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg 1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg 1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag 1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt 1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt 1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt 2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag 2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata 2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta 2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta 2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa 2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt 2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat 2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa 2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag 2581 agatccagtt tggttaatta atgcaaagat ggataaagtt ttaaacagag aggaatcttt 2641 gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt 2701 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa 2761 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc 2821 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc 2881 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc 2941 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccact ggctgcagta 3001 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc 3061 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg 3121 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat 3181 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc 3241 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg 3301 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg 3361 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat 3421 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg 3481 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg 3541 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg 3601 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag 3661 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg 3721 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct 3781 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct 3841 tccatttcag gtgtcgtgac gtacggccac catggctagc cccaaaaaga agaggaaagt 3901 gggatccgtg agtaaaggtg aagaactctt cactggagta gtgcccattc tggtagagct 3961 tgatggagat gtaaatggac ataaattctc cgtcaggggc gaaggcgaag gggacgccac 4021 gaatggtaag ctgactctga aattcatctg tacgacgggc aaactgcccg tcccatggcc 4081 tacactcgta acgaccctca cctacggcgt gcaatgcttt tctcgatatc ccgaccacat 4141 gaaacagcat gactttttca agtctgcaat gcctgaaggt tatgttcaag aaaggaccat 4201 cagctttaag gatgatggta catataaaac ccgagccgag gttaaatttg aaggggacac 4261 tctggttaat cgaattgaac tgaaaggtat tgattttaag gaggacggta acatactggg 4321 gcacaagttg gagtacaact ttaacagcca taatgtgtat attaccgctg ataagcagaa 4381 aaatgggata aaggccaact ttaagatccg acataatgtc gaagatggta gtgttcaact 4441 ggctgatcat taccaacaaa atacgcccat cggagatgga cctgtactct tgcctgacaa 4501 tcattatctc tccacgcaat caaagctttc caaggaccca aacgaaaaga gagatcacat 4561 ggtccttctg gaatttgtga ctgccgcagg catcactctc ggtatggatg agctgtacaa 4621 gggaggaggt ggaagcggag gaggaggaag cggaggagga ggtagcatga aagtgaccaa 4681 ggtcgacggc atcagccaca agaagtacat cgaagagggc aagctcgtga agtccaccag 4741 cgaggaaaac cggaccagcg agagactgag cgagctgctg agcatccggc tggacatcta 4801 catcaagaac cccgacaacg cctccgagga agagaaccgg atcagaagag agaacctgaa 4861 gaagttcttt agcaacaagg tgctgcacct gaaggacagc gtgctgtatc tgaagaaccg 4921 gaaagaaaag aacgccgtgc aggacaagaa ctatagcgaa gaggacatca gcgagtacga 4981 cctgaaaaac aagaacagct tctccgtgct gaagaagatc ctgctgaacg aggacgtgaa 5041 ctctgaggaa ctggaaatct ttcggaagga cgtggaagcc aagctgaaca agatcaacag 5101 cctgaagtac agcttcgaag agaacaaggc caactaccag aagatcaacg agaacaacgt 5161 ggaaaaagtg ggcggcaaga gcaagcggaa catcatctac gactactaca gagagagcgc 5221 caagcgcaac gactacatca acaacgtgca ggaagccttc gacaagctgt ataagaaaga 5281 ggatatcgag aaactgtttt tcctgatcga gaacagcaag aagcacgaga agtacaagat 5341 ccgcgagtac tatcacaaga tcatcggccg gaagaacgac aaagagaact tcgccaagat 5401 tatctacgaa gagatccaga acgtgaacaa catcaaagag ctgattgaga agatccccga 5461 catgtctgag ctgaagaaaa gccaggtgtt ctacaagtac tacctggaca aagaggaact 5521 gaacgacaag aatattaagt acgccttctg ccacttcgtg gaaatcgaga tgtcccagct 5581 gctgaaaaac tacgtgtaca agcggctgag caacatcagc aacgataaga tcaagcggat 5641 cttcgagtac cagaatctga aaaagctgat cgaaaacaaa ctgctgaaca agctggacac 5701 ctacgtgcgg aactgcggca agtacaacta ctatctgcaa gtgggcgaga tcgccacctc 5761 cgactttatc gcccggaacc ggcagaacga ggccttcctg agaaacatca tcggcgtgtc 5821 cagcgtggcc tacttcagcc tgaggaacat cctggaaacc gagaacgaga acggtatcac 5881 cggccggatg cggggcaaga ccgtgaagaa caacaagggc gaagagaaat acgtgtccgg 5941 cgaggtggac aagatctaca atgagaacaa gcagaacgaa gtgaaagaaa atctgaagat 6001 gttctacagc tacgacttca acatggacaa caagaacgag atcgaggact tcttcgccaa 6061 tatcgatgag gccatctcta gtattgctca cggcatcgtg catttcaacc tggaactgga 6121 aggcaaggac atcttcgcct tcaagaatat cgcccccagc gagatctcca agaagatgtt 6181 tcagaacgaa atcaacgaaa agaagctgaa gctgaaaatc ttcaagcagc tgaacagcgc 6241 caacgtgttc aactactacg agaaggatgt gatcatcaag tacctgaaga ataccaagtt 6301 caacttcgtg aacaaaaaca tccccttcgt gcccagcttc accaagctgt acaacaagat 6361 tgaggacctg cggaataccc tgaagttttt ttggagcgtg cccaaggaca aagaagagaa 6421 ggacgcccag atctacctgc tgaagaatat ctactacggc gagttcctga acaagttcgt 6481 gaaaaactcc aaggtgttct ttaagatcac caatgaagtg atcaagatta acaagcagcg 6541 gaaccagaaa accggccact acaagtatca gaagttcgag aacatcgaga aaaccgtgcc 6601 cgtggaatac ctggccatca tccagagcag agagatgatc aacaaccagg acaaagagga 6661 aaagaatacc tacatcgact ttattcagca gattttcctg aagggcttca tcgactacct 6721 gaacaagaac aatctgaagt atatcgagag caacaacaac aatgacaaca acgacatctt 6781 ctccaagatc aagatcaaaa aggataacaa agagaagtac gacaagatcc tgaagaacta 6841 tgagaagcac aatcggaaca aagaaatccc tcacgagatc aatgagttcg tgcgcgagat 6901 caagctgggg aagattctga agtacaccga gaatctgaac atgttttacc tgatcctgaa 6961 gctgctgaac cacaaagagc tgaccaacct gaagggcagc ctggaaaagt accagtccgc 7021 caacaaagaa gaaaccttca gcgacgagtt ggaactgatc aacctgctga acctggacaa 7081 caacagagtg accgaggact tcgagctgga agccaacgag atcggcaagt tcctggactt 7141 caacgaaaac aaaatcaagg accggaaaga gctgaaaaag ttcgacacca acaagatcta 7201 tttcgacggc gagaacatca tcaagcaccg ggccttctac aatatcaaga aatacggcat 7261 gctgaatctg ctggaaaaga tcgccgataa ggccaagtat aagatcagcc tgaaagaact 7321 gaaagagtac agcaacaaga agaatgagat tgaaaagaac tacaccatgc agcagaacct 7381 gcaccggaag tacgccagac ccaagaagga cgaaaagttc aacgacgagg actacaaaga 7441 gtatgagaag gccatcggca acatccagaa gtacacccac ctgaagaaca aggtggaatt 7501 caatgagctg aacctgctgc agggcctgct gctgaagatc ctgcaccggc tcgtgggcta 7561 caccagcatc tgggagcggg acctgagatt ccggctgaag ggcgagtttc ccgagaacca 7621 ctacatcgag gaaattttca atttcgacaa ctccaagaat gtgaagtaca aaagcggcca 7681 gatcgtggaa aagtatatca acttctacaa agaactgtac aaggacaatg tggaaaagcg 7741 gagcatctac tccgacaaga aagtgaagaa actgaagcag gagaagaaag acctgtatat 7801 tgcaaactac atcgctcact ttaattatat tcctcatgcc gagattagcc tgctggaagt 7861 gctggaaaac ctgcggaagc tgctgtccta cgaccggaag ctgaagaacg ccatcatgaa 7921 gtccatcgtg gacattctga aagaatacgg cttcgtggcc accttcaaga tcggcgctga 7981 caagaagatc gaaatccaga ccctggaatc agagaagatc gtgcacctga agaatctgaa 8041 gaaaaagaaa ctgatgaccg accggaacag cgaggaactg tgcgaactcg tgaaagtcat 8101 gttcgagtac aaggccctgg aaggatccga gggagctccc aagaaaaagc gcaaggtagg 8161 tagttcctac ccatacgatg ttccagatta cgcttatccc tacgacgtgc ctgattatgc 8221 atacccatat gatgtccccg actatgccta agaattcgat atcaagctta tcgataatca 8281 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt 8341 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc 8401 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc 8461 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg 8521 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc 8581 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg 8641 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg 8701 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc 8761 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct 8821 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc 8881 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga 8941 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt 9001 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa 9061 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat 9121 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag 9181 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga 9241 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga 9301 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat 9361 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg 9421 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt 9481 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc 9541 cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga 9601 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc 9661 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc 9721 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt 9781 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa 9841 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg 9901 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc 9961 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 10021 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 10081 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 10141 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 10201 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt 10261 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca 10321 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 10381 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 10441 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc 10501 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 10561 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt 10621 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga 10681 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag 10741 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac 10801 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag 10861 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac 10921 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac 10981 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc 11041 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc 11101 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc 11161 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 11221 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat 11281 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 11341 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg 11401 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 11461 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 11521 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 11581 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 11641 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 11701 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 11761 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 11821 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 11881 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 11941 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 12001 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 12061 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 12121 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 12181 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 12241 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg 12301 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 12361 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 12421 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 12481 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 12541 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 12601 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 12661 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 12721 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 12781 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 12841 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 12901 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 12961 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 13021 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 13081 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 13141 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 13201 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 13261 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 13321 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 13381 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 13441 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 13501 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 13561 tttccccgaa aagtgccacc tgac //