| 质粒名称: | pCEP4-M2L |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 20926 |
| 存储实验室: | James Thomson |
|
载体骨架基本信息 |
|
| 载体名称: | pCEP4 |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 10200 bp |
| 载体类型: | 染色体外/EBNA |
| 筛选标记: | 潮霉素 |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | MYC,bHLEHE39,c-Myc |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1300 bp |
| GenBank编号 | NM_002467.4 |
| 启动子 | CMV |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NheI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | IRES-R2 (gacggcaatatggtggaaa) |
|
文献引用方法 |
|
| 材料和方法部分: | pCEP4-M2L was a gift from James Thomson (Addgene plasmid # 20926 ; http://n2t.net/addgene:20926 ; RRID:Addgene_20926) |
| 参考文献部分: | Human Induced Pluripotent Stem Cells Free of Vector and Transgene Sequences. Yu J, Hu K, Smuga-Otto K, Tian S, Stewart R, Slukvin II, Thomson JA. Science. 2009 Mar 26. 10.1126/science.1172482 PubMed 19325077 |
买家导航
pCEP4M2L载体基本信息
pCEP4M2L载体图谱质粒图谱和多克隆位点信息
pCEP4M2L载体简介
pCEP4M2L载体序列
LOCUS pCEP4-M2L 12852 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION used in the derivation of human iPS cells using non-integrating episomal vectors; expresses cMyc and Lin28. ACCESSION . VERSION . KEYWORDS pCEP4-M2L SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12852) AUTHORS Yu J, Hu K, Smuga-Otto K, Tian S, Stewart R, Slukvin II, Thomson JA TITLE Human Induced Pluripotent Stem Cells Free of Vector and Transgene Sequences JOURNAL Science. 2009 Mar 26. PUBMED 19325077 REFERENCE 2 (bases 1 to 12852) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12852 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 4..383 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 384..587 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 538..558 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 584..608 /label=LNCX /note="Human CMV promoter, forward primer" CDS 683..2002 /codon_start=1 /product="human c-Myc proto-oncogene" /label=c-Myc /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA" misc_feature 2069..2642 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" misc_feature 2092..2644 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(2236..2253) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 2463..2482 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" polyA_signal 3338..3472 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3362..3381) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3416..3435 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(3615..3637) /label=pGEX 3' /note="pGEX vectors, reverse primer" rep_origin 4195..5984 /label=oriP /note="Epstein-Barr virus oriP replication origin (Yates et al., 2000)" CDS complement(6286..8211) /codon_start=1 /product="Epstein-Barr nuclear antigen 1, also known as EBNA-1" /label=EBNA1 /note="crucial for latent viral infection, and for episomal amplification of vectors containing the oriP origin (Okita et al., 2013)" /translation="MSDEGPGTGPGNGLGEKGDTSGPEGSGGSGPQRRGGDNHGRGRGR GRGRGGGRPGAPGGSGSGPRHRDGVRRPQKRPSCIGCKGTHGGTGAGAGAGGAGAGGAG AGGGAGAGGGAGGAGGAGGAGAGGGAGAGGGAGGAGGAGAGGGAGAGGGAGGAGAGGGA GGAGGAGAGGGAGAGGGAGGAGAGGGAGGAGGAGAGGGAGAGGAGGAGGAGAGGAGAGG GAGGAGGAGAGGAGAGGAGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGGAGAG GGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGAGGAGAGGGGRGRGGSGGRGRG GSGGRGRGGSGGRRGRGRERARGGSRERARGRGRGRGEKRPRSPSSQSSSSGSPPRRPP PGRRPFFHPVGEADYFEYHQEGGPDGEPDVPPGAIEQGPADDPGEGPSTGPRGQGDGGR RKKGGWFGKHRGQGGSNPKFENIAEGLRALLARSHVERTTDEGTWVAGVFVYGGSKTSL YNLRRGTALAIPQCRLTPLSRLPFGMAPGPGPQPGPLRESIVCYFMVFLQTHIFAEVLK DAIKDLVMTKPAPTCNIRVTVCSFDDGVDLPPWFPPMVEGAAAEGDDGDDGDEGGDGDE GEEGQE" primer_bind complement(8646..8664) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 8732..8836 /gene="bla" /label=AmpR promoter CDS 8837..9697 /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(9055..9074) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9868..10456 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 10357..10376 /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 10887..11032 /label=HSV TK promoter /note="herpes simplex virus thymidine kinase promoter" CDS 11075..12112 /codon_start=1 /product="hygromycin B phosphotransferase" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE MGEAN" primer_bind complement(12110..12129) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 12154..12202 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" ORIGIN 1 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 241 atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg 301 cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg 361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat 421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc 541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc 601 gtcagatctc tagaagctgg gtaccagctg ctagcgctac cggactcaga tctcgagctc 661 aagcttcgaa ttcgcgatcg ccatgcccct caacgttagc ttcaccaaca ggaactatga 721 cctcgactac gactcggtgc agccgtattt ctactgcgac gaggaggaga acttctacca 781 gcagcagcag cagagcgagc tgcagccccc ggcgcccagc gaggatatct ggaagaaatt 841 cgagctgctg cccaccccgc ccctgtcccc tagccgccgc tccgggctct gctcgccctc 901 ctacgttgcg gtcacaccct tctcccttcg gggagacaac gacggcggtg gcgggagctt 961 ctccacggcc gaccagctgg agatggtgac cgagctgctg ggaggagaca tggtgaacca 1021 gagtttcatc tgcgacccgg acgacgagac cttcatcaaa aacatcatca tccaggactg 1081 tatgtggagc ggcttctcgg ccgccgccaa gctcgtctca gagaagctgg cctcctacca 1141 ggctgcgcgc aaagacagcg gcagcccgaa ccccgcccgc ggccacagcg tctgctccac 1201 ctccagcttg tacctgcagg atctgagcgc cgccgcctca gagtgcatcg acccctcggt 1261 ggtcttcccc taccctctca acgacagcag ctcgcccaag tcctgcgcct cgcaagactc 1321 cagcgccttc tctccgtcct cggattctct gctctcctcg acggagtcct ccccgcaggg 1381 cagccccgag cccctggtgc tccatgagga gacaccgccc accaccagca gcgactctga 1441 ggaggaacaa gaagatgagg aagaaatcga tgttgtttct gtggaaaaga ggcaggctcc 1501 tggcaaaagg tcagagtctg gatcaccttc tgctggaggc cacagcaaac ctcctcacag 1561 cccactggtc ctcaagaggt gccacgtctc cacacatcag cacaactacg cagcgcctcc 1621 ctccactcgg aaggactatc ctgctgccaa gagggtcaag ttggacagtg tcagagtcct 1681 gagacagatc agcaacaacc gaaaatgcac cagccccagg tcctcggaca ccgaggagaa 1741 tgtcaagagg cgaacacaca acgtcttgga gcgccagagg aggaacgagc taaaacggag 1801 cttttttgcc ctgcgtgacc agatcccgga gttggaaaac aatgaaaagg cccccaaggt 1861 agttatcctt aaaaaagcca cagcatacat cctgtccgtc caagcagagg agcaaaagct 1921 catttctgaa gaggacttgt tgcggaaacg acgagaacag ttgaaacaca aacttgaaca 1981 gctacggaac tcttgtgcgt aagtttaaac atcgatggcc tcgagaaagc ggccgctttc 2041 tcgaggccat cgatgtttaa acgatccgcc cctctccctc ccccccccct aacgttactg 2101 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat 2161 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc 2221 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag 2281 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc 2341 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac 2401 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca 2461 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt 2521 gtatgggatc tgatctgggg cctcggtaca catgctttac atgtgtttag tcgaggttaa 2581 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat 2641 aatatggcca caccgcgatc gcgaattcga ttatcgccat gggctccgtg tccaaccagc 2701 agtttgcagg tggctgcgcc aaggcggcag aagaggcgcc cgaggaggcg ccggaggacg 2761 cggcccgggc ggcggacgag cctcagctgc tgcacggtgc gggcatctgt aagtggttca 2821 acgtgcgcat ggggttcggc ttcctgtcca tgaccgcccg cgccggggtc gcgctcgacc 2881 ccccagtgga tgtctttgtg caccagagta agctgcacat ggaagggttc cggagcttga 2941 aggagggtga ggcagtggag ttcaccttta agaagtcagc caagggtctg gaatccatcc 3001 gtgtcaccgg acctggtgga gtattctgta ttgggagtga gaggcggcca aaaggaaaga 3061 gcatgcagaa gcgcagatca aaaggagaca ggtgctacaa ctgtggaggt ctagatcatc 3121 atgccaagga atgcaagctg ccaccccagc ccaagaagtg ccacttctgc cagagcatca 3181 gccatatggt agcctcatgt ccgctgaagg cccagcaggg ccctagtgca cagggaaagc 3241 caacctactt tcgagaggaa gaagaagaaa tccacagccc taccctgctc ccggaggcac 3301 agaattgagt ttaaacnaat cactagctag ttcgaaggat ccagacatga taagatacat 3361 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat 3421 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa 3481 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa 3541 gtaaaacctc tacaaatgtg gtatggctga ttatgatccg gctgcctcgc gcgtttcggt 3601 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa 3661 gcggatgccg ggagcagaca agcccgtcag gcgtcagcgg gtgttggcgg gtgtcggggc 3721 gcagccatga ggtcgactct agaggatcga tgccccgccc cggacgaact aaacctgact 3781 acgacatctc tgccccttct tcgcggggca gtgcatgtaa tcccttcagt tggttggtac 3841 aacttgccaa ctgggccctg ttccacatgt gacacggggg gggaccaaac acaaaggggt 3901 tctctgactg tagttgacat ccttataaat ggatgtgcac atttgccaac actgagtggc 3961 tttcatcctg gagcagactt tgcagtctgt ggactgcaac acaacattgc ctttatgtgt 4021 aactcttggc tgaagctctt acaccaatgc tgggggacat gtacctccca ggggcccagg 4081 aagactacgg gaggctacac caacgtcaat cagaggggcc tgtgtagcta ccgataagcg 4141 gaccctcaag agggcattag caatagtgtt tataaggccc ccttgttaac cctaaacggg 4201 tagcatatgc ttcccgggta gtagtatata ctatccagac taaccctaat tcaatagcat 4261 atgttaccca acgggaagca tatgctatcg aattagggtt agtaaaaggg tcctaaggaa 4321 cagcgatatc tcccacccca tgagctgtca cggttttatt tacatggggt caggattcca 4381 cgagggtagt gaaccatttt agtcacaagg gcagtggctg aagatcaagg agcgggcagt 4441 gaactctcct gaatcttcgc ctgcttcttc attctccttc gtttagctaa tagaataact 4501 gctgagttgt gaacagtaag gtgtatgtga ggtgctcgaa aacaaggttt caggtgacgc 4561 ccccagaata aaatttggac ggggggttca gtggtggcat tgtgctatga caccaatata 4621 accctcacaa accccttggg caataaatac tagtgtagga atgaaacatt ctgaatatct 4681 ttaacaatag aaatccatgg ggtggggaca agccgtaaag actggatgtc catctcacac 4741 gaatttatgg ctatgggcaa cacataatcc tagtgcaata tgatactggg gttattaaga 4801 tgtgtcccag gcagggacca agacaggtga accatgttgt tacactctat ttgtaacaag 4861 gggaaagaga gtggacgccg acagcagcgg actccactgg ttgtctctaa cacccccgaa 4921 aattaaacgg ggctccacgc caatggggcc cataaacaaa gacaagtggc cactcttttt 4981 tttgaaattg tggagtgggg gcacgcgtca gcccccacac gccgccctgc ggttttggac 5041 tgtaaaataa gggtgtaata acttggctga ttgtaacccc gctaaccact gcggtcaaac 5101 cacttgccca caaaaccact aatggcaccc cggggaatac ctgcataagt aggtgggcgg 5161 gccaagatag gggcgcgatt gctgcgatct ggaggacaaa ttacacacac ttgcgcctga 5221 gcgccaagca cagggttgtt ggtcctcata ttcacgaggt cgctgagagc acggtgggct 5281 aatgttgcca tgggtagcat atactaccca aatatctgga tagcatatgc tatcctaatc 5341 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc 5401 tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc tatcctaatc 5461 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatc 5521 tgtatccggg tagcatatgc tatcctaata gagattaggg tagtatatgc tatcctaatt 5581 tatatctggg tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata 5641 tctgggtagc atatgctatc ctaatctata tctgggtagc ataggctatc ctaatctata 5701 tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc ctaatttata 5761 tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata 5821 tctgggtagt atatgctatc ctaatctgta tccgggtagc atatgctatc ctcatgcata 5881 tacagtcagc atatgatacc cagtagtaga gtgggagtgc tatcctttgc atatgccgcc 5941 acctcccaag ggggcgtgaa ttttcgctgc ttgtcctttt cctgctggtt gctcccattc 6001 ttaggtgaat ttaaggaggc caggctaaag ccgtcgcatg tctgattgct caccaggtaa 6061 atgtcgctaa tgttttccaa cgcgagaagg tgttgagcgc ggagctgagt gacgtgacaa 6121 catgggtatg cccaattgcc ccatgttggg aggacgaaaa tggtgacaag acagatggcc 6181 agaaatacac caacagcacg catgatgtct actggggatt tattctttag tgcgggggaa 6241 tacacggctt ttaatacgat tgagggcgtc tcctaacaag ttacatcact cctgcccttc 6301 ctcaccctca tctccatcac ctccttcatc tccgtcatct ccgtcatcac cctccgcggc 6361 agccccttcc accataggtg gaaaccaggg aggcaaatct actccatcgt caaagctgca 6421 cacagtcacc ctgatattgc aggtaggagc gggctttgtc ataacaaggt ccttaatcgc 6481 atccttcaaa acctcagcaa atatatgagt ttgtaaaaag accatgaaat aacagacaat 6541 ggactccctt agcgggccag gttgtgggcc gggtccaggg gccattccaa aggggagacg 6601 actcaatggt gtaagacgac attgtggaat agcaagggca gttcctcgcc ttaggttgta 6661 aagggaggtc ttactacctc catatacgaa cacaccggcg acccaagttc cttcgtcggt 6721 agtcctttct acgtgactcc tagccaggag agctcttaaa ccttctgcaa tgttctcaaa 6781 tttcgggttg gaacctcctt gaccacgatg ctttccaaac caccctcctt ttttgcgcct 6841 gcctccatca ccctgacccc ggggtccagt gcttgggcct tctcctgggt catctgcggg 6901 gccctgctct atcgctcccg ggggcacgtc aggctcacca tctgggccac cttcttggtg 6961 gtattcaaaa taatcggctt cccctacagg gtggaaaaat ggccttctac ctggaggggg 7021 cctgcgcggt ggagacccgg atgatgatga ctgactactg ggactcctgg gcctcttttc 7081 tccacgtcca cgacctctcc ccctggctct ttcacgactt ccccccctgg ctctttcacg 7141 tcctctaccc cggcggcctc cactacctcc tcgaccccgg cctccactac ctcctcgacc 7201 ccggcctcca ctgcctcctc gaccccggcc tccacctcct gctcctgccc ctcctgctcc 7261 tgcccctcct cctgctcctg cccctcctgc ccctcctgct cctgcccctc ctgcccctcc 7321 tgctcctgcc cctcctgccc ctcctgctcc tgcccctcct gcccctcctc ctgctcctgc 7381 ccctcctgcc cctcctcctg ctcctgcccc tcctgcccct cctgctcctg cccctcctgc 7441 ccctcctgct cctgcccctc ctgcccctcc tgctcctgcc cctcctgctc ctgcccctcc 7501 tgctcctgcc cctcctgctc ctgcccctcc tgcccctcct gcccctcctc ctgctcctgc 7561 ccctcctgct cctgcccctc ctgcccctcc tgcccctcct gctcctgccc ctcctcctgc 7621 tcctgcccct cctgcccctc ctgcccctcc tcctgctcct gcccctcctg cccctcctcc 7681 tgctcctgcc cctcctcctg ctcctgcccc tcctgcccct cctgcccctc ctcctgctcc 7741 tgcccctcct gcccctcctc ctgctcctgc ccctcctcct gctcctgccc ctcctgcccc 7801 tcctgcccct cctcctgctc ctgcccctcc tcctgctcct gcccctcctg cccctcctgc 7861 ccctcctgcc cctcctcctg ctcctgcccc tcctcctgct cctgcccctc ctgctcctgc 7921 ccctcccgct cctgctcctg ctcctgttcc accgtgggtc cctttgcagc caatgcaact 7981 tggacgtttt tggggtctcc ggacaccatc tctatgtctt ggccctgatc ctgagccgcc 8041 cggggctcct ggtcttccgc ctcctcgtcc tcgtcctctt ccccgtcctc gtccatggtt 8101 atcaccccct cttctttgag gtccactgcc gccggagcct tctggtccag atgtgtctcc 8161 cttctctcct aggccatttc caggtcctgt acctggcccc tcgtcagaca tgattcacac 8221 taaaagagat caatagacat ctttattaga cgacgctcag tgaatacagg gagtgcagac 8281 tcctgccccc tccaacagcc cccccaccct catccccttc atggtcgctg tcagacagat 8341 ccaggtctga aaattcccca tcctccgaac catcctcgtc ctcatcacca attactcgca 8401 gcccggaaaa ctcccgctga acatcctcaa gatttgcgtc ctgagcctca agccaggcct 8461 caaattcctc gtcccccttt ttgctggacg gtagggatgg ggattctcgg gacccctcct 8521 cttcctcttc aaggtcacca gacagagatg ctactggggc aacggaagaa aagctgggtg 8581 cggcctgtga ggatcagctt atcgatgata agctgtcaaa catgagaatt cttgaagacg 8641 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta 8701 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 8761 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 8821 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc 8881 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga 8941 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct 9001 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg 9061 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta 9121 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat 9181 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt 9241 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga 9301 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga 9361 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga 9421 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc 9481 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc 9541 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg 9601 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat 9661 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata 9721 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 9781 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 9841 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 9901 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 9961 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct 10021 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 10081 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 10141 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 10201 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 10261 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 10321 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 10381 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 10441 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 10501 gccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc 10561 gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg 10621 cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc ccgagatgcg ccgcgtgcgg 10681 ctgctggaga tggcggacgc gatggatatg ttctgccaag ggttggtttg cgcattcaca 10741 gttctccgca agaattgatt ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc 10801 cgccctgctt catccccgtg gcccgttgct cgcgtttgct ggcggtgtcc ccggaagaaa 10861 tatatttgca tgtctttagt tctatgatga cacaaacccc gcccagcgtc ttgtcattgg 10921 cgaattcgaa cacgcagatg cagtcggggc ggcgcggtcc gaggtccact tcgcatatta 10981 aggtgacgcg tgtggcctcg aacaccgagc gaccctgcag cgacccgctt aacagcgtca 11041 acagcgtgcc gcagatcccg gggggcaatg agatatgaaa aagcctgaac tcaccgcgac 11101 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc 11161 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg 11221 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc 11281 ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta 11341 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc 11401 cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca 11461 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga 11521 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac 11581 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc 11641 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg 11701 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt 11761 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt 11821 cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat 11881 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc 11941 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat 12001 cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg 12061 aaaccgacgc cccagcactc gtccggatcg ggagatgggg gaggctaact gaaacacgga 12121 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc 12181 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc 12241 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc 12301 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct 12361 gccatagcca ctggccccgt gggttaggga cggggtcccc catggggaat ggtttatggt 12421 tcgtgggggt tattattttg ggcgttgcgt ggggtcaggt ccacgactgg actgagcaga 12481 cagacccatg gtttttggat ggcctgggca tggaccgcat gtactggcgc gacacgaaca 12541 ccgggcgtct gtggctgcca aacacccccg acccccaaaa accaccgcgc ggatttctgg 12601 cgtgccaagc tagtcgacca attctcatgt ttgacagctt atcatcgcag atccgggcaa 12661 cgttgttgcc attgctgcag gcgcagaact ggtaggtatg gaagatctat acattgaatc 12721 aatattggca attagccata ttagtcattg gttatatagc ataaatcaat attggctatt 12781 ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa 12841 tatgaccgcc at //
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