质粒名称: | pEBTet_NADP-Snifit-cyto |
---|---|
出品公司: | 爱迪基因 |
目录编号: | 113917 |
存储实验室: | Kai Johnsson |
载体骨架基本信息 |
|
载体名称: | pEBTet |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | 11448 bp |
完整质粒大小: | 13797 bp |
载体修饰: | -- |
载体类型: | 哺乳细胞表达 |
筛选标记: | 嘌呤霉素 |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
|
插入基因名称: | NADP-Snifit,hSPR-HaloTag7-Pro30-SNAPtag |
物种来源: | Synthetic |
插入大小: | 2349 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | CMV TetO2 |
标签/融合蛋白: | -- |
克隆方法: | Gibson克隆 |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | CMV_Fwd |
3' 测序引物 | BGH_Rev |
文献引用方法 |
|
材料和方法部分: | pEBTet_NADP-Snifit-cyto was a gift from Kai Johnsson (Addgene plasmid # 113917 ; http://n2t.net/addgene:113917 ; RRID:Addgene_113917) |
参考文献部分: | Semisynthetic biosensors for mapping cellular concentrations of nicotinamide adenine dinucleotides. Sallin O, Reymond L, Gondrand C, Raith F, Koch B, Johnsson K. Elife. 2018 May 29;7. pii: 32638. doi: 10.7554/eLife.32638. 32638 [pii] PubMed 29809136 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pEBTet_NADP-Snifit-cyto 13797 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Plasmid encoding the cytosolic localization of the FRET semi-synthetic biosensor measuring the ratio NADPH/NADP+ in mammalian cells. ACCESSION . VERSION . KEYWORDS pEBTet_NADP-Snifit-cyto SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13797) AUTHORS Sallin O, Reymond L, Gondrand C, Raith F, Koch B, Johnsson K TITLE Semisynthetic biosensors for mapping cellular concentrations of nicotinamide adenine dinucleotides. JOURNAL Elife. 2018 May 29;7. pii: 32638. doi: 10.7554/eLife.32638. PUBMED 29809136 REFERENCE 2 (bases 1 to 13797) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13797 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 807..1691 /codon_start=1 /product="modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives" /label=HaloTag(R) /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG SEIARWLSTLEISG" primer_bind complement(2422..2439) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2428..2539 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3080..4869 /label=oriP /note="Epstein-Barr virus oriP replication origin (Yates et al., 2000)" primer_bind complement(7533..7551) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 7619..7723 /gene="bla" /label=AmpR promoter CDS 7724..8584 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(7942..7961) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8755..9343 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9244..9263 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind complement(9532..9552) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 9550..9847 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind 9779..9798 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 9924..10523 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(9924..9943) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 10420..10440 /label=Puro-F /note="Puromycin resistance gene, forward primer" polyA_signal 10614..10748 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(10886..10905) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 11182..11443 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" intron 11560..12132 /label=-beta-globin intron /note="intron from rabbit beta-globin gene" primer_bind 12140..12159 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" primer_bind 12187..12206 /label=T7 /note="T7 promoter, forward primer" promoter 12187..12205 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 12216..12833 /codon_start=1 /gene="tetR from transposon Tn10" /product="tetracycline repressor TetR" /label=TetR /note="TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline." /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG" primer_bind complement(12289..12308) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" polyA_signal 12878..12999 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" enhancer 13054..13433 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 13434..13637 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 13588..13608 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 13639..13657 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 13660..13678 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 13688..13712 /label=LNCX /note="Human CMV promoter, forward primer" ORIGIN 1 agcttaccac catggagggc gggctggggc gtgctgtgtg cttgctgacc ggggcctccc 61 gcggcttcgg ccggacgctg gccccgctcc tggcctcgct gctgtcgccc ggctccgtgc 121 ttgtccttag cgcccgcaac gacgaggcac tgcgccagct ggaggccgag ctgggcgccg 181 agcggtctgg cctgcgcgtg gtgcgggtgc ccgccgacct gggcgccgag gccggcttgc 241 agcagctgct cggcgccctg cgcgagctcc cccggcccaa ggggctgcag cgactgctgc 301 ttatcaacaa cgcgggctct cttggggatg tgtccaaagg cttcgtggac ctgagtgact 361 ccactcaagt gaacaactac tgggcactga acttgacctc catgctctgc ctgacttcca 421 gcgtcctgaa ggccttcccg gacagtcctg gcctcaacag aaccgtggtt aacatctcgt 481 ccctctgtgc cctgcaacct ttcaaaggct gggcgctgta ctgtgcagga aaggctgctc 541 gtgatatgct gttccaggtc ctggcgctgg aggaacctaa tgtgagggtg ctgaactatg 601 ccccaggtcc tctggacaca gacatgcagc agttggcccg ggagacctcc gtggacccag 661 acatgcgaaa agggctgcag gagctgaagg caaaggggaa gctggtggat tgcaaggtgt 721 cagcccagaa actgctgagc ttactggaaa aggacgagtt caagtctgga gcccacgtgg 781 acttctatga caaagaattc ggatccgaaa tcggtactgg ctttccattc gacccccatt 841 atgtggaagt cctgggcgag cgcatgcact acgtcgatgt tggtccgcgc gatggcaccc 901 ctgtgctgtt cctgcacggt aacccgacct cctcctacgt gtggcgcaac atcatcccgc 961 atgttgcacc gacccatcgc tgcattgctc cagacctgat cggtatgggc aaatccgaca 1021 aaccagacct gggttatttc ttcgacgacc acgtccgctt catggatgcc ttcatcgaag 1081 ccctgggtct ggaagaggtc gtcctggtca ttcacgactg gggctccgct ctgggtttcc 1141 actgggccaa gcgcaatcca gagcgcgtca aaggtattgc atttatggag ttcatccgcc 1201 ctatcccgac ctgggacgaa tggccagaat ttgcccgcga gaccttccag gccttccgca 1261 ccaccgacgt cggccgcaag ctgatcatcg atcagaacgt ttttatcgag ggtacgctgc 1321 cgatgggtgt cgtccgcccg ctgactgaag tcgagatgga ccattaccgc gagccgttcc 1381 tgaatcctgt tgaccgcgag ccactgtggc gcttcccaaa cgagctgcca atcgccggtg 1441 agccagcgaa catcgtcgcg ctggtcgaag aatacatgga ctggctgcac cagtcccctg 1501 tcccgaagct gctgttctgg ggcaccccag gcgttctgat cccaccggcc gaagccgctc 1561 gcctggccaa aagcctgcct aactgcaagg ctgtggacat cggcccgggt ctgaatctgc 1621 tgcaagaaga caacccggac ctgatcggca gcgagatcgc gcgctggctg tctactctgg 1681 agatttccgg tagcggccgc cctcctcctc cgccaccgcc accaccgccg ccgccgccac 1741 caccacctcc tccgcctcct ccaccaccac cgccaccgcc tccaccacct ggcggccgct 1801 ctcggtctct cgagatggat aaagactgcg aaatgaaacg cactaccctg gacagcccgc 1861 tgggtaaact ggaactgagc ggttgcgagc agggcctgca cgaaatcatt ttcctgggca 1921 aaggtacgag cgctgcggat gcggtcgaag taccggctcc ggctgcggtt ctgggtggcc 1981 cggagccgct gatgcaggct accgcatggc tgaacgcata ttttcaccag ccagaggcga 2041 ttgaagagtt ccctgtaccg gccctgcatc acccggtatt tcagcaggaa tcctttaccc 2101 gtcaggtact gtggaaactg ctgaaagtag tgaaattcgg cgaggtgatc tcctattccc 2161 atctggcggc gctggctggc aacccagcgg caactgcggc ggtgaaaacc gctctgtctg 2221 gtaatccggt gccaattctg atcccgtgcc accgtgtggt tcagggcgac ctggacgtag 2281 gtggctacga aggcggcctg gctgtgaaag aatggctgct ggcgcacgaa ggtcaccgtc 2341 tgggtaaacc aggcctgggc taaagctagc ggcgcgcccg gccgctcgag tctagagggc 2401 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt 2461 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat 2521 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg 2581 tggggcagga cagcaagggg gaggattggg aagacaatag caggctagag gatcgatccc 2641 cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca 2701 tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac 2761 ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg 2821 tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact 2881 gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg 2941 gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag 3001 gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa 3061 ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc 3121 cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta 3181 gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt 3241 ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt 3301 ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct 3361 ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc 3421 tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt 3481 ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg 3541 taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg 3601 taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg 3661 caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat 3721 gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc 3781 actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa 3841 acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc 3901 cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta 3961 accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg 4021 aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg 4081 acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac 4141 gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat 4201 ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat 4261 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat 4321 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat 4381 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat 4441 tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg 4501 atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg 4561 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg 4621 gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg 4681 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg 4741 gtagcatatg ctatcctcat gcatatacag tcagcatatg atacccagta gtagagtggg 4801 agtgctatcc tttgcatatg ccgccacctc ccaagggggc gtgaattttc gctgcttgtc 4861 cttttcctgc tggttgctcc cattcttagg tgaatttaag gaggccaggc taaagccgtc 4921 gcatgtctga ttgctcacca ggtaaatgtc gctaatgttt tccaacgcga gaaggtgttg 4981 agcgcggagc tgagtgacgt gacaacatgg gtatgcccaa ttgccccatg ttgggaggac 5041 gaaaatggtg acaagacaga tggccagaaa tacaccaaca gcacgcatga tgtctactgg 5101 ggatttattc tttagtgcgg gggaatacac ggcttttaat acgattgagg gcgtctccta 5161 acaagttaca tcactcctgc ccttcctcac cctcatctcc atcacctcct tcatctccgt 5221 catctccgtc atcaccctcc gcggcagccc cttccaccat aggtggaaac cagggaggca 5281 aatctactcc atcgtcaaag ctgcacacag tcaccctgat attgcaggta ggagcgggct 5341 ttgtcataac aaggtcctta atcgcatcct tcaaaacctc agcaaatata tgagtttgta 5401 aaaagaccat gaaataacag acaatggact cccttagcgg gccaggttgt gggccgggtc 5461 caggggccat tccaaagggg agacgactca atggtgtaag acgacattgt ggaatagcaa 5521 gggcagttcc tcgccttagg ttgtaaaggg aggtcttact acctccatat acgaacacac 5581 cggcgaccca agttccttcg tcggtagtcc tttctacgtg actcctagcc aggagagctc 5641 ttaaaccttc tgcaatgttc tcaaatttcg ggttggaacc tccttgacca cgatgctttt 5701 ccaaaccacc ctcctttttt gcgccctgcc tccatcaccc tgaccccggg gtccagtgct 5761 tgggccttct cctgggtcat ctgcggggcc ctgctctatc gctcccgggg gcacgtcagg 5821 ctcaccatct gggccacctt cttggtggta ttcaaaataa tcggcttccc ctacagggtg 5881 gaaaaatggc cttctacctg gagggggcct gcgcggtgga gacccggatg atgatgactg 5941 actactggga ctcctgggcc tcttttctcc acgtccacga cctctccccc tggctctttc 6001 acgacttccc cccctggctc tttcacgtcc tctaccccgg cggcctccac tacctcctcg 6061 accccggcct ccactacctc ctcgaccccg gcctccactg cctcctcgac cccggcctcc 6121 acctcctgct cctgcccctc ctgctcctgc ccctcctcct gctcctgccc ctcctgcccc 6181 tcctgctcct gcccctcctg cccctcctgc tcctgcccct cctgcccctc ctgctcctgc 6241 ccctcctgcc cctcctcctg ctcctgcccc tcctgcccct cctcctgctc ctgcccctcc 6301 tgcccctcct gctcctgccc ctcctgcccc tcctgctcct gcccctcctg cccctcctgc 6361 tcctgcccct cctgctcctg cccctcctgc tcctgcccct cctgctcctg cccctcctgc 6421 ccctcctgcc cctcctcctg ctcctgcccc tcctgctcct gcccctcctg cccctcctgc 6481 ccctcctgct cctgcccctc ctcctgctcc tgcccctcct gcccctcctg cccctcctcc 6541 tgctcctgcc cctcctgccc ctcctcctgc tcctgcccct cctcctgctc ctgcccctcc 6601 tgcccctcct gcccctcctc ctgctcctgc ccctcctgcc cctcctcctg ctcctgcccc 6661 tcctcctgct cctgcccctc ctgcccctcc tgcccctcct cctgctcctg cccctcctcc 6721 tgctcctgcc cctcctgccc ctcctgcccc tcctgcccct cctcctgctc ctgcccctcc 6781 tcctgctcct gcccctcctg ctcctgcccc tcccgctcct gctcctgctc ctgttccacc 6841 gtgggtccct ttgcagccaa tgcaacttgg acgtttttgg ggtctccgga caccatctct 6901 atgtcttggc cctgatcctg agccgcccgg ggctcctggt cttccgcctc ctcgtcctcg 6961 tcctcttccc cgtcctcgtc catggttatc accccctctt ctttgaggtc cactgccgcc 7021 ggagccttct ggtccagatg tgtctccctt ctctcctagg ccatttccag gtcctgtacc 7081 tggcccctcg tcagacatga ttcacactaa aagagatcaa tagacatctt tattagacga 7141 cgctcagtga atacagggag tgcagactcc tgccccctcc aacagccccc ccaccctcat 7201 ccccttcatg gtcgctgtca gacagatcca ggtctgaaaa ttccccatcc tccgaaccat 7261 cctcgtcctc atcaccaatt actcgcagcc cggaaaactc ccgctgaaca tcctcaagat 7321 ttgcgtcctg agcctcaagc caggcctcaa attcctcgtc cccctttttg ctggacggta 7381 gggatgggga ttctcgggac ccctcctctt cctcttcaag gtcaccagac agagatgcta 7441 ctggggcaac ggaagaaaag ctgggtgcgg cctgtgagga tcagcttatc gatgataagc 7501 tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt 7561 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg 7621 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca 7681 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt 7741 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga 7801 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga 7861 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat 7921 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca 7981 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt 8041 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac 8101 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct 8161 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga 8221 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac 8281 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat 8341 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg 8401 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc 8461 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc 8521 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg 8581 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta 8641 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg 8701 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga 8761 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt 8821 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag 8881 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa 8941 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag 9001 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca 9061 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac 9121 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa 9181 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc 9241 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg 9301 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc 9361 ctttttacgg ttcctggcct tttgctggcc ttgaagctgt ccctgatggt cgtcatctac 9421 ctgcctggac agcatggcct gcaacgcggg catcccgatg ccgccggaag cgagaagaat 9481 cataatgggg aaggccatcc agcctcgcgt cgaactagat gatccggctg tggaatgtgt 9541 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 9601 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 9661 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 9721 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 9781 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct 9841 tttttggagg gtgaccgcca cgaccggtgc cgccaccatc ccctgaccca cgcccctgac 9901 ccctcacaag gagacgacct tccatgaccg agtacaagcc cacggtgcgc ctcgccaccc 9961 gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca 10021 cgcgccacac cgtcgacccc gaccgccaca tcgaacgcgt caccgagctg caagaactct 10081 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg 10141 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc 10201 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc 10261 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg 10321 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc 10381 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc 10441 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca 10501 tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga 10561 gcgcacgacc cggtccgacg cggcccgacg ggtccgaggg gggtcgacct cgaaacttgt 10621 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag 10681 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg 10741 tctggatctg atccgaaccc cttcctcgac caattctcat gtttgacagc ttatcatcgc 10801 agatccgggc aacgttgttg ccattgctcg accaattctc atgtttgaca gcttatcatc 10861 gcagatccgt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt 10921 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta 10981 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg 11041 cgctgcttcg cgatgtacgg gccagatata cgcgtatctg aggggactag ggtgtgttta 11101 ggcgaaaagc ggggcttcgg ttgtacgcgg ttaggagtcc cctcaggata tagtagtttc 11161 gcttttgcat agggaggggg aaatgtagtc ttatgcaata ctcttgtagt cttgcaacat 11221 ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt 11281 ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg 11341 gattggacga accactgaat tccgcattgc agagatattg tatttaagtg cctagctcga 11401 tacaataaac gccatttgac cattcaccac attggtgtgc acctccaagc tggccgggga 11461 tcctctaggg cctctgagct attccagaag tagtgaagag gcttttttgg aggcctaggc 11521 ttttgcaaaa agctccggat cgatcctgag aacttcaggg tgagtttggg gacccttgat 11581 tgttctttct ttttcgctat tgtaaaattc atgttatatg gagggggcaa agttttcagg 11641 gtgttgttta gaatgggaag atgtcccttg tatcaccatg gaccctcatg ataattttgt 11701 ttctttcact ttctactctg ttgacaacca ttgtctcctc ttattttctt ttcattttct 11761 gtaacttttt cgttaaactt tagcttgcat ttgtaacgaa tttttaaatt cacttttgtt 11821 tatttgtcag attgtaagta ctttctctaa tcactttttt ttcaaggcaa tcagggtata 11881 ttatattgta cttcagcaca gttttagaga acaattgtta taattaaatg ataaggtaga 11941 atatttctgc atataaattc tggctggcgt ggaaatattc ttattggtag aaacaactac 12001 atcctggtca tcatcctgcc tttctcttta tggttacaat gatatacact gtttgagatg 12061 aggataaaat actctgagtc caaaccgggc ccctctgcta accatgttca tgccttcttc 12121 tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa 12181 gaattgtaat acgactcact atagggcgaa ttgatatgtc tagattagat aaaagtaaag 12241 tgattaacag cgcattagag ctgcttaatg aggtcggaat cgaaggttta acaacccgta 12301 aactcgccca gaagctaggt gtagagcagc ctacattgta ttggcatgta aaaaataagc 12361 gggctttgct cgacgcctta gccattgaga tgttagatag gcaccatact cacttttgcc 12421 ctttagaagg ggaaagctgg caagattttt tacgtaataa cgctaaaagt tttagatgtg 12481 ctttactaag tcatcgcgat ggagcaaaag tacatttagg tacacggcct acagaaaaac 12541 agtatgaaac tctcgaaaat caattagcct ttttatgcca acaaggtttt tcactagaga 12601 atgcattata tgcactcagc gctgtggggc attttacttt aggttgcgta ttggaagatc 12661 aagagcatca agtcgctaaa gaagaaaggg aaacacctac tactgatagt atgccgccat 12721 tattacgaca agctatcgaa ttatttgatc accaaggtgc agagccagcc ttcttattcg 12781 gccttgaatt gatcatatgc ggattagaaa aacaacttaa atgtgaaagt gggtccgcgt 12841 acagcggatc ccgggaattc agatcttatt aaagcagaac ttgtttattg cagcttataa 12901 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 12961 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggt cgaatccatg 13021 gcgcagaact ggtaggtatg gaagatccgc gttgacattg attattgact agttattaat 13081 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac 13141 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa 13201 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt 13261 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc 13321 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat 13381 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc 13441 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc 13501 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa 13561 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg 13621 tctatataag cagagctctc cctatcagtg atagagatct ccctatcagt gatagagatc 13681 gtcgacgagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg 13741 acctccatag aagacaccgg gaccgatcca gcctccggac tctagcgttt aaactta //