| 质粒名称: | pEP4 E02S CK2M EN2L |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 20924 |
| 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: | -- |
| 完整质粒大小: | 11600 bp |
| 载体修饰: | -- |
| 载体类型: | -- |
| 筛选标记: | 新霉素（G418） |
| 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: | Oct4 and Sox2; Nanog and Lin28; Klf4 and Myc |
| 物种来源: | -- |
| 插入大小: | 4900 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pEP4 E02S CK2M EN2L was a gift from James Thomson (Addgene plasmid # 20924 ; http://n2t.net/addgene:20924 ; RRID:Addgene_20924) |
| 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
pEP4 E02S CK2M EN2L载体质粒基本信息
pEP4 E02S CK2M EN2L质粒图谱载体图谱和pEP4 E02S CK2M EN2L载体序列质粒序列多克隆位点信息
pEP4 E02S CK2M EN2L质粒载体简介
pEP4 E02S CK2M EN2L质粒序列载体序列
LOCUS pEP4 E02S CK2M EN2L 19949 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION used in the derivation of human iPS cells using non-integrating
episomal vectors; expresses Oct4 and Sox2; Klf4 and Myc; Nanog and
Lin28.
ACCESSION .
VERSION .
KEYWORDS pEP4 E02S CK2M EN2L
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 19949)
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 19949)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..19949
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 13..1175
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 244..1166
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
misc_feature 2385..2958
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
misc_feature 2408..2960
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
polyA_signal 3977..4111
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
enhancer 4618..4997
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 4998..5201
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 5152..5172
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
misc_feature 6713..7286
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
misc_feature 6736..7288
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 7348..8667
/codon_start=1
/product="human c-Myc proto-oncogene"
/label=c-Myc
/translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP
SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE
LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN
PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS
LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS
PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR
KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA
TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA"
primer_bind complement(8724..8741)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 8730..8954
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
promoter 9175..10337
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 9406..10328
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
misc_feature 11381..11954
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
misc_feature 11404..11956
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
polyA_signal 12650..12784
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 13515..15304
/label=oriP
/note="Epstein-Barr virus oriP replication origin (Yates et
al., 2000)"
CDS complement(15606..17531)
/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(17970..17988)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 18056..18160
/gene="bla"
/label=AmpR promoter
CDS 18161..19021
/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(18379..18398)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 19192..19780
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 19681..19700
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ctagcttcgt gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
61 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta
121 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
181 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
241 caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc
301 gtgccttgaa ttacttccac ctggctccag tacgtgattc ttgatcccga gctggagcca
361 ggggcgggcc ttgcgcttta ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg
421 ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg
481 ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag
541 atagtcttgt aaatgcgggc caggatctgc acactggtat ttcggttttt gggcccgcgg
601 ccggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc
661 ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc
721 tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg
781 cgtgagcgga aagatggccg cttcccggcc ctgctccagg gggctcaaaa tggaggacgc
841 ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct
901 cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt
961 tctggagctt ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt
1021 ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct
1081 cgttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg
1141 ttcaaagttt ttttcttcca tttcaggtgt cgtgaacacg tggtcgcggc caattctgca
1201 gtcgagctca agcttcgaac tagtgattat ggcgggacac ctggcttcgg atttcgcctt
1261 ctcgccccct ccaggtggtg gaggtgatgg gccagggggg ccggagccgg gctgggttga
1321 tcctcggacc tggctaagct tccaaggccc tcctggaggg ccaggaatcg ggccgggggt
1381 tgggccaggc tctgaggtgt gggggattcc cccatgcccc ccgccgtatg agttctgtgg
1441 ggggatggcg tactgtgggc cccaggttgg agtggggcta gtgccccaag gcggcttgga
1501 gacctctcag cctgagggtg aagcangagt cggggtggag agcaactccg atggggcctc
1561 cccggagccc tgcaccgtca cccctggtgc cgtgaagctg gagaaggaga agctggagca
1621 aaacccggag gagtcccagg acatcaaagc tctgcagaaa gaactcgagc aatttgccaa
1681 gctcctgaag cagaagagga tcaccctggg atatacacag gccgatgtgg ggctcaccct
1741 gggggttcta tttgggaagg tattcagcca aacgaccatc tgccgctttg aggctctgca
1801 gcttagcttc aagaacatgt gtaagctgcg gcccttgctg cagaagtggg tggaggaagc
1861 tgacaacaat gaaaatcttc aggagatatg caaagcagaa accctcgtgc aggcccgaaa
1921 gagaaagcga accagtatcg agaaccgagt gagaggcaac ctggagaatt tgttcctgca
1981 gtgcccgaaa cccacactgc agcagatcag ccacatcgcc cagcagcttg ggctcgagaa
2041 ggatgtggtc cgagtgtggt tctgtaaccg gcgccagaag ggcaagcgat caagcagcga
2101 ctatgcacaa cgagaggatt ttgaggctgc tgggtctcct ttctcagggg gaccagtgtc
2161 ctttcctctg gccccagggc cccattttgg taccccaggc tatgggagcc ctcacttcac
2221 tgcactgtac tcctcggtcc ctttccctga gggggaagcc tttccccctg tctccgtcac
2281 cactctgggc tctcccatgc attcaaactg aggtgcctgc cctctagttt acgcgtgggg
2341 gcggccggcc gctttctcga ggccatcgat gtttaaacga tccgcccctc tccctccccc
2401 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
2461 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
2521 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
2581 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
2641 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
2701 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
2761 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
2821 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtacacatg ctttacatgt
2881 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
2941 tgaaaaacac gatgataata tggccacacc gcgaattaat tcgattccgc atgtacaaca
3001 tgatggagac ggagctgaag ccgccgggcc cgcagcaaac ttcggggggc ggcggcggca
3061 actccaccgc ggcggcggcc ggcggcaacc agaaaaacag cccggaccgc gtcaagcggc
3121 ccatgaatgc cttcatggtg tggtcccgcg ggcagcggcg caagatggcc caggagaacc
3181 ccaagatgca caactcggag atcagcaagc gcctgggcgc cgagtggaaa cttttgtcgg
3241 agacggagaa gcggccgttc atcgacgagg ctaagcggct gcgagcgctg cacatgaagg
3301 agcacccgga ttataaatac cggccccggc ggaaaaccaa gacgctcatg aagaaggata
3361 agtacacgct gcccggcggg ctgctggccc ccggcggcaa tagcatggcg agcggggtcg
3421 gggtgggcgc cggcctgggc gcgggcgtga accagcgcat ggacagttac gcgcacatga
3481 acggctggag caacggcagc tacagcatga tgcaggacca gctgggctac ccgcagcacc
3541 cgggcctcaa tgcgcacggc gcagcgcaga tgcagcccat gcaccgctac gacgtgagcg
3601 ccctgcagta caactccatg accagctcgc agacctacat gaacggctcg cccacctaca
3661 gcatgtccta ctcgcagcag ggcacccctg gcatggctct tggctccatg ggttcggtgg
3721 tcaagtccga ggccagctcc agcccccctg tggttacctc ttcctcccac tccagggcgc
3781 cctgccaggc cggggacctc cgggacatga tcagcatgta tctccccggc gccgaggtgc
3841 cggaacccgc cgcccccagc agacttcaca tgtcccagca ctaccagagc ggcccggtgc
3901 ccggcacggc cattaacggc acactgcccc tctcacacat gtgagggccg gaaatcacta
3961 gctagttcga aggatcgatc cagacatgat aagatacatt gatgagtttg gacaaaccac
4021 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
4081 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
4141 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
4201 tatggctgat tatgatccgg ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
4261 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
4321 gcccgtcagg cgtcagcggg tgttggcggg tgtcggggcg cagccatgag gtcgaccaat
4381 tctcatgttt gacagcttat catcgcagat ccgggcaacg ttgttgccat tgctgcaggc
4441 gcagaactgg taggtatgga agatcgatct atacattgaa tcaatattgg caattagcca
4501 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
4561 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac
4621 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
4681 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
4741 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
4801 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
4861 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
4921 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
4981 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
5041 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
5101 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
5161 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
5221 tccgctagga tctcgagctc aagcttcgaa ttcgcgatcg ccatggctgt cagcgacgcg
5281 ctgctcccat ctttctccac gttcgcgtct ggcccggcgg gaagggagaa gacactgcgt
5341 caagcaggtg ccccgaataa ccgctggcgg gaggagctct cccacatgaa gcgacttccc
5401 ccagtgcttc ccggccgccc ctatgacctg gcggcggcga ccgtggccac agacctggag
5461 agcggcggag ccggtgcggc ttgcggcggt agcaacctgg cgcccctacc tcggagagag
5521 accgaggagt tcaacgatct cctggacctg gactttattc tctccaattc gctgacccat
5581 cctccggagt cagtggccgc caccgtgtcc tcgtcagcgt cagcctcctc ttcgtcgtcg
5641 ccgtcgagca gcggccctgc cagcgcgccc tccacctgca gcttcaccta tccgatccgg
5701 gccgggaacg acccgggcgt ggcgccgggc ggcacgggcg gaggcctcct ctatggcagg
5761 gagtccgctc cccctccgac ggctcccttc aacctggcgg acatcaacga cgtgagcccc
5821 tcgggcggct tcatggccga gctcctgcgg ccagaattgg acccggtgta cattccgccg
5881 cagcagccgc agccgccagg tggcgggctg atgggcaagt tcgtgctgaa ggcgtcgctg
5941 agcgcccctg gcagcgagta cggcagcccg tcggtcatca gcgtcagcaa aggcagccct
6001 gacggcagcc acccggtggt ggtggcgccc tacaacggcg ggccgccgcg cacgtgcccc
6061 aagatcaagc aggaggcggt ctcttcgtgc acccacttgg gcgctggacc ccctctcagc
6121 aatggccacc ggccggctgc acacgacttc cccctggggc ggcagctccc cagcaggact
6181 accccgaccc tgggtcttga ggaagtgctg agcagcaggg actgtcaccc tgccctgccg
6241 cttcctcccg gcttccatcc ccacccgggg cccaattacc catccttcct gcccgatcag
6301 atgcagccgc aagtcccgcc gctccattac caagagctca tgccacccgg ttcctgcatg
6361 ccagaggagc ccaagccaaa gaggggaaga cgatcgtggc cccggaaaag gaccgccacc
6421 cacacttgtg attacgcggg ctgcggcaaa acctacacaa agagttccca tctcaaggca
6481 cacctgcgaa cccacacagg tgagaaacct taccactgtg actgggacgg ctgtggatgg
6541 aaattcgccc gctcagatga actgaccagg cactaccgta aacacacggg gcaccgcccg
6601 ttccagtgcc aaaaatgcga ccgagcattt tccaggtcgg accacctcgc cttacacatg
6661 aagaggcatt tttaaatccc agacagtgga tatgacccgt ttaaacgatc cgcccctctc
6721 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
6781 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
6841 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
6901 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
6961 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
7021 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
7081 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
7141 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct
7201 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
7261 ttttcctttg aaaaacacga tgataatatg gccacaccgc gatcgcgaat tctgcagtcg
7321 agctcaagct tcgaattcgc gatcgccatg cccctcaacg ttagcttcac caacaggaac
7381 tatgacctcg actacgactc ggtgcagccg tatttctact gcgacgagga ggagaacttc
7441 taccagcagc agcagcagag cgagctgcag cccccggcgc ccagcgagga tatctggaag
7501 aaattcgagc tgctgcccac cccgcccctg tcccctagcc gccgctccgg gctctgctcg
7561 ccctcctacg ttgcggtcac acccttctcc cttcggggag acaacgacgg cggtggcggg
7621 agcttctcca cggccgacca gctggagatg gtgaccgagc tgctgggagg agacatggtg
7681 aaccagagtt tcatctgcga cccggacgac gagaccttca tcaaaaacat catcatccag
7741 gactgtatgt ggagcggctt ctcggccgcc gccaagctcg tctcagagaa gctggcctcc
7801 taccaggctg cgcgcaaaga cagcggcagc ccgaaccccg cccgcggcca cagcgtctgc
7861 tccacctcca gcttgtacct gcaggatctg agcgccgccg cctcagagtg catcgacccc
7921 tcggtggtct tcccctaccc tctcaacgac agcagctcgc ccaagtcctg cgcctcgcaa
7981 gactccagcg ccttctctcc gtcctcggat tctctgctct cctcgacgga gtcctccccg
8041 cagggcagcc ccgagcccct ggtgctccat gaggagacac cgcccaccac cagcagcgac
8101 tctgaggagg aacaagaaga tgaggaagaa atcgatgttg tttctgtgga aaagaggcag
8161 gctcctggca aaaggtcaga gtctggatca ccttctgctg gaggccacag caaacctcct
8221 cacagcccac tggtcctcaa gaggtgccac gtctccacac atcagcacaa ctacgcagcg
8281 cctccctcca ctcggaagga ctatcctgct gccaagaggg tcaagttgga cagtgtcaga
8341 gtcctgagac agatcagcaa caaccgaaaa tgcaccagcc ccaggtcctc ggacaccgag
8401 gagaatgtca agaggcgaac acacaacgtc ttggagcgcc agaggaggaa cgagctaaaa
8461 cggagctttt ttgccctgcg tgaccagatc ccggagttgg aaaacaatga aaaggccccc
8521 aaggtagtta tccttaaaaa agccacagca tacatcctgt ccgtccaagc agaggagcaa
8581 aagctcattt ctgaagagga cttgttgcgg aaacgacgag aacagttgaa acacaaactt
8641 gaacagctac ggaactcttg tgcgtaagtt tggccgcccc cacgcgtaaa ctagttcgaa
8701 ggatcctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
8761 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
8821 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
8881 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
8941 ggtgggctct atggcttctg aggcggaaag aaccagctgg gggcgcgccc ctcgagatct
9001 atacattgaa tcaatattgg caattagcca tattagtcat tggttatata gcataaatca
9061 atattggcta ttggccattg catacgttgt atctatatca taatatgtac atttatattg
9121 gctcatgtcc aatatgaccg ccatgttgac attgattatt gactagcttc gtgaggctcc
9181 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
9241 gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
9301 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
9361 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt
9421 ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc
9481 acctggctcc agtacgtgat tcttgatccc gagctggagc caggggcggg ccttgcgctt
9541 taggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
9601 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
9661 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
9721 gccaggatct gcacactggt atttcggttt ttgggcccgc ggccggcgac ggggcccgtg
9781 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
9841 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg
9901 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
9961 cgcttcccgg ccctgctcca gggggctcaa aatggaggac gcggcgctcg ggagagcggg
10021 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
10081 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctggagc ttttggagta
10141 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
10201 tggagactga agttaggcca gcttggcact tgatgtaatt ctcgttggaa tttgcccttt
10261 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
10321 catttcaggt gtcgtgaaca cgtggtcgcg gccaattctg cagtcgagct caagcttcga
10381 actagggccg cgactctagt gattcatgag tgtggatcca gcttgtcccc aaagcttgcc
10441 ttgctttgaa gcatccgact gtaaagaatc ttcacctatg cctgtgattt gtgggcctga
10501 agaaaactat ccatccttgc aaatgtcttc tgctgagatg cctcacacgg agactgtctc
10561 tcctcttcct tcctccatgg atctgcttat tcaggacagc cctgattctt ccaccagtcc
10621 caaaggcaaa caacccactt ctgcagagaa gagtgtcgca aaaaaggaag acaaggtccc
10681 ggtcaagaaa cagaagacca gaactgtgtt ctcttccacc cagctgtgtg tactcaatga
10741 tagatttcag agacagaaat acctcagcct ccagcagatg caagaactct ccaacatcct
10801 gaacctcagc tacaaacagg tgaagacctg gttccagaac cagagaatga aatctaagag
10861 gtggcagaaa aacaactggc cgaagaatag caatggtgtg acgcagaagg cctcagcacc
10921 tacctacccc agcctttact cttcctacca ccagggatgc ctggtgaacc cgactgggaa
10981 ccttccaatg tggagcaacc agacctggaa caattcaacc tggagcaacc agacccagaa
11041 catccagtcc tggagcaacc actcctggaa cactcagacc tggtgcaccc aatcctggaa
11101 caatcaggcc tggaacagtc ccttctataa ctgtggagag gaatctctgc agtcctgcat
11161 gcagttccag ccaaattctc ctgccagtga cttggaggct gccttggaag ctgctgggga
11221 aggccttaat gtaatacagc agaccactag gtattttagt actccacaaa ccatggattt
11281 attcctaaac tactccatga acatgcaacc tgaagacgtg tgaagatgaa tcgaattccc
11341 gcggccgctt tctcgaggcc atcgatgttt aaacgatccg cccctctccc tccccccccc
11401 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
11461 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
11521 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
11581 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
11641 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
11701 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
11761 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
11821 aggtacccca ttgtatggga tctgatctgg ggcctcggta cacatgcttt acatgtgttt
11881 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
11941 aaacacgatg ataatatggc cacaccgcga tcgcgaattc gattatcgcc atgggctccg
12001 tgtccaacca gcagtttgca ggtggctgcg ccaaggcggc agaagaggcg cccgaggagg
12061 cgccggagga cgcggcccgg gcggcggacg agcctcagct gctgcacggt gcgggcatct
12121 gtaagtggtt caacgtgcgc atggggttcg gcttcctgtc catgaccgcc cgcgccgggg
12181 tcgcgctcga ccccccagtg gatgtctttg tgcaccagag taagctgcac atggaagggt
12241 tccggagctt gaaggagggt gaggcagtgg agttcacctt taagaagtca gccaagggtc
12301 tggaatccat ccgtgtcacc ggacctggtg gagtattctg tattgggagt gagaggcggc
12361 caaaaggaaa gagcatgcag aagcgcagat caaaaggaga caggtgctac aactgtggag
12421 gtctagatca tcatgccaag gaatgcaagc tgccacccca gcccaagaag tgccacttct
12481 gccagagcat cagccatatg gtagcctcat gtccgctgaa ggcccagcag ggccctagtg
12541 cacagggaaa gccaacctac tttcgagagg aagaagaaga aatccacagc cctaccctgc
12601 tcccggaggc acagaattga gtttaaacna atcactagct agttcgaagg atccagacat
12661 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
12721 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
12781 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
12841 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc cggctgcctc
12901 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
12961 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc aggcgtcagc gggtgttggc
13021 gggtgtcggg gcgcagccat gaggtcgatc gactctagct agaggatcga tgccccgccc
13081 cggacgaact aaacctgact acgacatctc tgccccttct tcgcggggca gtgcatgtaa
13141 tcccttcagt tggttggtac aacttgccaa ctgggccctg ttccacatgt gacacggggg
13201 gggaccaaac acaaaggggt tctctgactg tagttgacat ccttataaat ggatgtgcac
13261 atttgccaac actgagtggc tttcatcctg gagcagactt tgcagtctgt ggactgcaac
13321 acaacattgc ctttatgtgt aactcttggc tgaagctctt acaccaatgc tgggggacat
13381 gtacctccca ggggcccagg aagactacgg gaggctacac caacgtcaat cagaggggcc
13441 tgtgtagcta ccgataagcg gaccctcaag agggcattag caatagtgtt tataaggccc
13501 ccttgttaac cctaaacggg tagcatatgc ttcccgggta gtagtatata ctatccagac
13561 taaccctaat tcaatagcat atgttaccca acgggaagca tatgctatcg aattagggtt
13621 agtaaaaggg tcctaaggaa cagcgatatc tcccacccca tgagctgtca cggttttatt
13681 tacatggggt caggattcca cgagggtagt gaaccatttt agtcacaagg gcagtggctg
13741 aagatcaagg agcgggcagt gaactctcct gaatcttcgc ctgcttcttc attctccttc
13801 gtttagctaa tagaataact gctgagttgt gaacagtaag gtgtatgtga ggtgctcgaa
13861 aacaaggttt caggtgacgc ccccagaata aaatttggac ggggggttca gtggtggcat
13921 tgtgctatga caccaatata accctcacaa accccttggg caataaatac tagtgtagga
13981 atgaaacatt ctgaatatct ttaacaatag aaatccatgg ggtggggaca agccgtaaag
14041 actggatgtc catctcacac gaatttatgg ctatgggcaa cacataatcc tagtgcaata
14101 tgatactggg gttattaaga tgtgtcccag gcagggacca agacaggtga accatgttgt
14161 tacactctat ttgtaacaag gggaaagaga gtggacgccg acagcagcgg actccactgg
14221 ttgtctctaa cacccccgaa aattaaacgg ggctccacgc caatggggcc cataaacaaa
14281 gacaagtggc cactcttttt tttgaaattg tggagtgggg gcacgcgtca gcccccacac
14341 gccgccctgc ggttttggac tgtaaaataa gggtgtaata acttggctga ttgtaacccc
14401 gctaaccact gcggtcaaac cacttgccca caaaaccact aatggcaccc cggggaatac
14461 ctgcataagt aggtgggcgg gccaagatag gggcgcgatt gctgcgatct ggaggacaaa
14521 ttacacacac ttgcgcctga gcgccaagca cagggttgtt ggtcctcata ttcacgaggt
14581 cgctgagagc acggtgggct aatgttgcca tgggtagcat atactaccca aatatctgga
14641 tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg
14701 tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg
14761 tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg
14821 tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctaata gagattaggg
14881 tagtatatgc tatcctaatt tatatctggg tagcatatac tacccaaata tctggatagc
14941 atatgctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagc
15001 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
15061 atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc
15121 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc
15181 atatgctatc ctcatgcata tacagtcagc atatgatacc cagtagtaga gtgggagtgc
15241 tatcctttgc atatgccgcc acctcccaag ggggcgtgaa ttttcgctgc ttgtcctttt
15301 cctgctggtt gctcccattc ttaggtgaat ttaaggaggc caggctaaag ccgtcgcatg
15361 tctgattgct caccaggtaa atgtcgctaa tgttttccaa cgcgagaagg tgttgagcgc
15421 ggagctgagt gacgtgacaa catgggtatg cccaattgcc ccatgttggg aggacgaaaa
15481 tggtgacaag acagatggcc agaaatacac caacagcacg catgatgtct actggggatt
15541 tattctttag tgcgggggaa tacacggctt ttaatacgat tgagggcgtc tcctaacaag
15601 ttacatcact cctgcccttc ctcaccctca tctccatcac ctccttcatc tccgtcatct
15661 ccgtcatcac cctccgcggc agccccttcc accataggtg gaaaccaggg aggcaaatct
15721 actccatcgt caaagctgca cacagtcacc ctgatattgc aggtaggagc gggctttgtc
15781 ataacaaggt ccttaatcgc atccttcaaa acctcagcaa atatatgagt ttgtaaaaag
15841 accatgaaat aacagacaat ggactccctt agcgggccag gttgtgggcc gggtccaggg
15901 gccattccaa aggggagacg actcaatggt gtaagacgac attgtggaat agcaagggca
15961 gttcctcgcc ttaggttgta aagggaggtc ttactacctc catatacgaa cacaccggcg
16021 acccaagttc cttcgtcggt agtcctttct acgtgactcc tagccaggag agctcttaaa
16081 ccttctgcaa tgttctcaaa tttcgggttg gaacctcctt gaccacgatg ctttccaaac
16141 caccctcctt ttttgcgcct gcctccatca ccctgacccc ggggtccagt gcttgggcct
16201 tctcctgggt catctgcggg gccctgctct atcgctcccg ggggcacgtc aggctcacca
16261 tctgggccac cttcttggtg gtattcaaaa taatcggctt cccctacagg gtggaaaaat
16321 ggccttctac ctggaggggg cctgcgcggt ggagacccgg atgatgatga ctgactactg
16381 ggactcctgg gcctcttttc tccacgtcca cgacctctcc ccctggctct ttcacgactt
16441 ccccccctgg ctctttcacg tcctctaccc cggcggcctc cactacctcc tcgaccccgg
16501 cctccactac ctcctcgacc ccggcctcca ctgcctcctc gaccccggcc tccacctcct
16561 gctcctgccc ctcctgctcc tgcccctcct cctgctcctg cccctcctgc ccctcctgct
16621 cctgcccctc ctgcccctcc tgctcctgcc cctcctgccc ctcctgctcc tgcccctcct
16681 gcccctcctc ctgctcctgc ccctcctgcc cctcctcctg ctcctgcccc tcctgcccct
16741 cctgctcctg cccctcctgc ccctcctgct cctgcccctc ctgcccctcc tgctcctgcc
16801 cctcctgctc ctgcccctcc tgctcctgcc cctcctgctc ctgcccctcc tgcccctcct
16861 gcccctcctc ctgctcctgc ccctcctgct cctgcccctc ctgcccctcc tgcccctcct
16921 gctcctgccc ctcctcctgc tcctgcccct cctgcccctc ctgcccctcc tcctgctcct
16981 gcccctcctg cccctcctcc tgctcctgcc cctcctcctg ctcctgcccc tcctgcccct
17041 cctgcccctc ctcctgctcc tgcccctcct gcccctcctc ctgctcctgc ccctcctcct
17101 gctcctgccc ctcctgcccc tcctgcccct cctcctgctc ctgcccctcc tcctgctcct
17161 gcccctcctg cccctcctgc ccctcctgcc cctcctcctg ctcctgcccc tcctcctgct
17221 cctgcccctc ctgctcctgc ccctcccgct cctgctcctg ctcctgttcc accgtgggtc
17281 cctttgcagc caatgcaact tggacgtttt tggggtctcc ggacaccatc tctatgtctt
17341 ggccctgatc ctgagccgcc cggggctcct ggtcttccgc ctcctcgtcc tcgtcctctt
17401 ccccgtcctc gtccatggtt atcaccccct cttctttgag gtccactgcc gccggagcct
17461 tctggtccag atgtgtctcc cttctctcct aggccatttc caggtcctgt acctggcccc
17521 tcgtcagaca tgattcacac taaaagagat caatagacat ctttattaga cgacgctcag
17581 tgaatacagg gagtgcagac tcctgccccc tccaacagcc cccccaccct catccccttc
17641 atggtcgctg tcagacagat ccaggtctga aaattcccca tcctccgaac catcctcgtc
17701 ctcatcacca attactcgca gcccggaaaa ctcccgctga acatcctcaa gatttgcgtc
17761 ctgagcctca agccaggcct caaattcctc gtcccccttt ttgctggacg gtagggatgg
17821 ggattctcgg gacccctcct cttcctcttc aaggtcacca gacagagatg ctactggggc
17881 aacggaagaa aagctgggtg cggcctgtga ggatcagctt atcgatgata agctgtcaaa
17941 catgagaatt aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
18001 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
18061 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
18121 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
18181 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
18241 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
18301 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
18361 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
18421 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
18481 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
18541 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
18601 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
18661 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
18721 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
18781 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
18841 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
18901 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
18961 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
19021 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
19081 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
19141 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
19201 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
19261 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
19321 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
19381 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
19441 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
19501 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
19561 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
19621 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
19681 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
19741 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
19801 tttacggttc ctggcctttt gctggccttg aagctgtccc tgatggtcgt catctacctg
19861 cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat
19921 aatggggaag gccatccagc ctcgcgtcg
//
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pEP4 E02S CK2M EN2L质粒载体应用举例
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