LOCUS Exported 11195 bp ds-DNA circular SYN 22-OCT-2021 DEFINITION CAG promoter expression plasmid for human codon optimized BE4max C-to-T base editor with SpRY(D10A/A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R133 3P/R1335Q/T1337R) and P2A-EGFP. ACCESSION . VERSION . KEYWORDS pCAG-CBE4max-SpRY-P2A-EGFP (RTW5133) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11195) AUTHORS Walton RT, Christie KA, Whittaker MN, Kleinstiver BP TITLE Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants. JOURNAL Science. 2020 Apr 17;368(6488):290-296. doi: 10.1126/science.aba8853. Epub 2020 Mar 26. PUBMED 32217751 REFERENCE 2 (bases 1 to 11195) AUTHORS . TITLE Direct Submission JOURNAL Exported Oct 22, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11195 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 37..416 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 418..693 /label=chicken beta-actin promoter intron 694..1710 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1634..1656 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1718..1737 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 1845..1865 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1866..2549 /codon_start=1 /product="cytidine deaminase (C to U editing enzyme) from rat" /label=APOBEC-1 /note="can use ssDNA as a substrate (Komor et al., 2016)" /translation="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL WATGLK" CDS 2646..4361 /codon_start=1 /product="N-terminal portion of Streptococcus pyogenes Cas9 (Zetsche et al., 2015)" /label=Cas9(N) /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAERTRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE" CDS 6777..7025 /codon_start=1 /product="uracil-DNA glycosylase inhibitor from a Bacillus subtilis bacteriophage (Mol et al., 1995)" /label=UGI /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML" CDS 7056..7304 /codon_start=1 /product="uracil-DNA glycosylase inhibitor from a Bacillus subtilis bacteriophage (Mol et al., 1995)" /label=UGI /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML" CDS 7347..7367 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 7386..7442 /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 7443..8159 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(7488..7509) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(7749..7768) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 8096..8117 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(8249..8268) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 8314..8369 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(8368..8387) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(8730..8746) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8730..8746) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(8743..8765) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8754..8770 /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(8778..8808) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8823..8844 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 8903..9098 /label=SV40 promoter /note="SV40 early promoter" rep_origin 8949..9084 /label=SV40 ori /note="SV40 origin of replication" primer_bind 9011..9030 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 9104..9238 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(9141..9160) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 9195..9214 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(9306..9323) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(9477..10065) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(9557..9576) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(10236..11096) /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 10859..10878 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(11097..6) /gene="bla" /label=AmpR promoter ORIGIN 1 tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca ttgattattg actagttatt 61 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat 121 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa 181 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg 241 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc 301 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct 361 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg 421 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt 481 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggcgc 541 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg 601 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg 661 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc 721 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc 781 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa 841 tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct 901 ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg 961 tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg 1021 ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc 1081 tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg 1141 ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc 1201 ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg 1261 gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg 1321 ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc 1381 cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt 1441 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg 1501 tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt 1561 ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga 1621 cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac 1681 catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg 1741 tctcatcatt ttggcaaaga attctgcagt cgacggtacc gcgggcccgg gatccaccgg 1801 tcgccaccat gaaacggaca gccgacggaa gcgagttcga gtcaccaaag aagaagcgga 1861 aagtctcctc agagactggg cctgtcgccg tcgatccaac cctgcgccgc cggattgaac 1921 ctcacgagtt tgaagtgttc tttgaccccc gggagctgag aaaggagaca tgcctgctgt 1981 acgagatcaa ctggggaggc aggcactcca tctggaggca cacctctcag aacacaaata 2041 agcacgtgga ggtgaacttc atcgagaagt ttaccacaga gcggtacttc tgccccaata 2101 ccagatgtag catcacatgg tttctgagct ggtccccttg cggagagtgt agcagggcca 2161 tcaccgagtt cctgtccaga tatccacacg tgacactgtt tatctacatc gccaggctgt 2221 atcaccacgc agacccaagg aataggcagg gcctgcgcga tctgatcagc tccggcgtga 2281 ccatccagat catgacagag caggagtccg gctactgctg gcggaacttc gtgaattatt 2341 ctcctagcaa cgaggcccac tggcctaggt acccacacct gtgggtgcgc ctgtacgtgc 2401 tggagctgta ttgcatcatc ctgggcctgc ccccttgtct gaatatcctg cggagaaagc 2461 agccccagct gaccttcttt acaatcgccc tgcagtcttg tcactatcag aggctgccac 2521 cccacatcct gtgggccaca ggcctgaagt ctggaggatc tagcggagga tcctctggca 2581 gcgagacacc aggaacaagc gagtcagcaa caccagagag cagtggcggc agcagcggcg 2641 gcagcgacaa gaagtacagc atcggcctgg ccatcggcac caactctgtg ggctgggccg 2701 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc 2761 ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg 2821 agagaacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct 2881 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca 2941 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg 3001 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa 3061 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc 3121 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg 3181 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc 3241 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca 3301 gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa 3361 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg 3421 aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg 3481 cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca 3541 tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct 3601 ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc 3661 ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg 3721 ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc 3781 tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc 3841 ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc 3901 acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga 3961 tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca 4021 gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg 4081 aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata 4141 agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg 4201 tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc 4261 tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga 4321 ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa 4381 tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa 4441 ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg 4501 tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg 4561 cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg 4621 gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc 4681 tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg 4741 acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc 4801 tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga 4861 cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg 4921 tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga 4981 gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc 5041 ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc 5101 gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc 5161 atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa 5221 gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga 5281 agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc 5341 tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac 5401 agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga 5461 acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt 5521 ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca 5581 actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa 5641 agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga 5701 agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca 5761 gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc 5821 ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt 5881 ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg 5941 tgcagacagg cggcttcagc aaagagtcta tcagacccaa gaggaacagc gataagctga 6001 tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcctgtgg cccaccgtgg 6061 cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg 6121 tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg 6181 actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta 6241 agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccaagcagc 6301 tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca 6361 gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg 6421 aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag 6481 tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata 6541 agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc agactgggag 6601 cccctagagc cttcaagtac tttgacacca ccatcgaccc caagcagtac agaagcacca 6661 aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga 6721 tcgacctgtc tcagctggga ggtgacagcg gcgggagcgg cgggagcggg gggagcacta 6781 atctgagcga catcattgag aaggagactg ggaaacagct ggtcattcag gagtccatcc 6841 tgatgctgcc tgaggaggtg gaggaagtga tcggcaacaa gccagagtct gacatcctgg 6901 tgcacaccgc ctacgacgag tccacagatg agaatgtgat gctgctgacc tctgacgccc 6961 ccgagtataa gccttgggcc ctggtcatcc aggattctaa cggcgagaat aagatcaaga 7021 tgctgagcgg aggatccgga ggatctggag gcagcaccaa cctgtctgac atcatcgaga 7081 aggagacagg caagcagctg gtcatccagg agagcatcct gatgctgccc gaagaagtcg 7141 aagaagtgat cggaaacaag cctgagagcg atatcctggt ccataccgcc tacgacgaga 7201 gtaccgacga aaatgtgatg ctgctgacat ccgacgcccc agagtataag ccctgggctc 7261 tggtcatcca ggattccaac ggagagaaca aaatcaaaat gctgtctggc ggctcaaaaa 7321 gaaccgccga cggcagcgaa ttcgagccca agaagaagag gaaagtcgga ggcggtggaa 7381 gcggcgcaac aaacttctct ctgctgaaac aagccggaga tgtcgaagag aatcctggac 7441 cgatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg 7501 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct 7561 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca 7621 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga 7681 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct 7741 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc 7801 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc 7861 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga 7921 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg 7981 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc 8041 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg 8101 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt 8161 gaagcggccg cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca 8221 atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca 8281 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa 8341 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt 8401 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg 8461 ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc 8521 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg 8581 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt 8641 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg 8701 acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 8761 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 8821 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 8881 aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa 8941 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 9001 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt 9061 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc 9121 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 9181 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg 9241 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 9301 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 9361 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 9421 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 9481 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 9541 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 9601 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 9661 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 9721 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 9781 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 9841 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 9901 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 9961 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 10021 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 10081 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 10141 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 10201 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 10261 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 10321 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 10381 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 10441 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 10501 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 10561 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 10621 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 10681 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 10741 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 10801 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 10861 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 10921 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 10981 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 11041 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 11101 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 11161 tttgaatgta tttagaaaaa taaacaaata ggggt //