| 质粒名称: | pCX-OKS-2A |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 19771 |
| 存储实验室: | Shinya Yamanaka |
|
载体骨架基本信息 |
|
| 载体名称: | pCX |
| 载体出品公司: | Jun-ichi Miyazaki |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4778 bp |
| 载体类型: | 哺乳细胞表达 |
| 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: | Oct3/4-2A-Klf4-2A-Sox2,Pou5f1,Sox2,Klf4 |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 3700 bp |
| 基因突变: | Three genes are connected with the foot-and-mouth disease virus 2A self-cleaving sequence. |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 5' 克隆位点: | EcoRI(未破坏) |
| 3' 克隆位点: | EcoRI (未破坏) |
| 5' 测序引物 | GCGAGCCGCAGCCATTGCCTTTTA |
| 3' 测序引物 | TTAGCCAGAAGTCAGATGCTC |
|
文献引用方法 |
|
| 材料和方法部分: | pCX-OKS-2A was a gift from Shinya Yamanaka (Addgene plasmid # 19771 ; http://n2t.net/addgene:19771 ; RRID:Addgene_19771) |
| 参考文献部分: | Generation of Mouse Induced Pluripotent Stem Cells Without Viral Vectors. Okita K, Nakagawa M, Hyenjong H, Ichisaka T, Yamanaka S. Science. 2008 Oct 9. ():. 10.1126/science.1164270 PubMed 18845712 |
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 pCX-OKS-2A 8495 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Non-integrating (episomal) expression of mouse Oct3/4, Klf4 and
Sox2.
ACCESSION .
VERSION .
KEYWORDS pCX-OKS-2A
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8495)
AUTHORS Okita K, Nakagawa M, Hyenjong H, Ichisaka T, Yamanaka S
TITLE Generation of Mouse Induced Pluripotent Stem Cells Without Viral
Vectors.
JOURNAL Science. 2008 Oct 9. ():.
PUBMED 18845712
REFERENCE 2 (bases 1 to 8495)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8495
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 4..383
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 385..661
/label=chicken beta-actin promoter
intron 662..1678
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1602..1624
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1686..1705
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 2842..2907
/codon_start=1
/product="2A peptide from foot-and-mouth disease virus
polyprotein"
/label=F2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="VKQTLNFDLLKLAGDVESNPGP"
CDS 4357..4422
/codon_start=1
/product="2A peptide from foot-and-mouth disease virus
polyprotein"
/label=F2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="VKQTLNFDLLKLAGDVESNPGP"
primer_bind complement(5516..5535)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 5581..5636
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(5635..5654)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(5997..6013)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5997..6013)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6010..6032)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6021..6037
/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(6045..6075)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6090..6111
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6170..6365
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 6216..6351
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 6278..6297
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 6371..6505
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6408..6427)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6462..6481
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(6573..6590)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6744..7332)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6824..6843)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7503..8363)
/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 8126..8145
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8364..8468)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacattg 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 agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
481 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
541 ggggcgggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgcccttaa
1741 aaagcaggct ccaccttccc catggctgga cacctggctt cagacttcgc cttctcaccc
1801 ccaccaggtg ggggtgatgg gtcagcaggg ctggagccgg gctgggtgga ttctcgaacc
1861 tggctaagct tccaagggcc tccaggtggg cctggaatcg gaccaggctc agaggtattg
1921 gggatctccc catgtccgcc cgcatacgag ttctgcggag ggatggcata ctgtggacct
1981 caggttggac tgggcctagt cccccaagtt ggcgtggaga ctttgcagcc tgagggccag
2041 gcaggagcac gagtggaaag caactcagag ggaacctcct ctgagccctg tgccgaccgc
2101 cccaatgccg tgaagttgga gaaggtggaa ccaactcccg aggagtccca ggacatgaaa
2161 gccctgcaga aggagctaga acagtttgcc aagctgctga agcagaagag gatcaccttg
2221 gggtacaccc aggccgacgt ggggctcacc ctgggcgttc tctttggaaa ggtgttcagc
2281 cagaccacca tctgtcgctt cgaggccttg cagctcagcc ttaagaacat gtgtaagctg
2341 cggcccctgc tggagaagtg ggtggaggaa gccgacaaca atgagaacct tcaggagata
2401 tgcaaatcgg agaccctggt gcaggcccgg aagagaaagc gaactagcat tgagaaccgt
2461 gtgaggtgga gtctggagac catgtttctg aagtgcccga agccctccct acagcagatc
2521 actcacatcg ccaatcagct tgggctagag aaggatgtgg ttcgagtatg gttctgtaac
2581 cggcgccaga agggcaaaag atcaagtatt gagtattccc aacgagaaga gtatgaggct
2641 acagggacac ctttcccagg gggggctgta tcctttcctc tgcccccagg tccccacttt
2701 ggcaccccag gctatggaag cccccacttc accacactct actcagtccc ttttcctgag
2761 ggcgaggcct ttccctctgt tcccgtcact gctctgggct ctcccatgca ttcaaacggg
2821 gatctcaaaa ttgtcgctcc tgtcaaacaa actcttaact ttgatttact caaactggct
2881 ggggatgtag aaagcaatcc aggtccactc atggctgtca gcgacgctct gctcccgtcc
2941 ttctccacgt tcgcgtccgg cccggcggga agggagaaga cactgcgtcc agcaggtgcc
3001 ccgactaacc gttggcgtga ggaactctct cacatgaagc gacttccccc acttcccggc
3061 cgcccctacg acctggcggc gacggtggcc acagacctgg agagtggcgg agctggtgca
3121 gcttgcagca gtaacaaccc ggccctccta gcccggaggg agaccgagga gttcaacgac
3181 ctcctggacc tagactttat cctttccaac tcgctaaccc accaggaatc ggtggccgcc
3241 accgtgacca cctcggcgtc agcttcatcc tcgtcttccc cagcgagcag cggccctgcc
3301 agcgcgccct ccacctgcag cttcagctat ccgatccggg ccgggggtga cccgggcgtg
3361 gctgccagca acacaggtgg agggctcctc tacagccgag aatctgcgcc acctcccacg
3421 gcccccttca acctggcgga catcaatgac gtgagcccct cgggcggctt cgtggctgag
3481 ctcctgcggc cggagttgga cccagtatac attccgccac agcagcctca gccgccaggt
3541 ggcgggctga tgggcaagtt tgtgctgaag gcgtctctga ccacccctgg cagcgagtac
3601 agcagccctt cggtcatcag tgttagcaaa ggaagcccag acggcagcca ccccgtggta
3661 gtggcgccct acagcggtgg cccgccgcgc atgtgcccca agattaagca agaggcggtc
3721 ccgtcctgca cggtcagccg gtccctagag gcccatttga gcgctggacc ccagctcagc
3781 aacggccacc ggcccaacac acacgacttc cccctggggc ggcagctccc caccaggact
3841 acccctacac tgagtcccga ggaactgctg aacagcaggg actgtcaccc tggcctgcct
3901 cttcccccag gattccatcc ccatccgggg cccaactacc ctcctttcct gccagaccag
3961 atgcagtcac aagtcccctc tctccattat caagagctca tgccaccggg ttcctgcctg
4021 ccagaggagc ccaagccaaa gaggggaaga aggtcgtggc cccggaaaag aacagccacc
4081 cacacttgtg actatgcagg ctgtggcaaa acctatacca agagttctca tctcaaggca
4141 cacctgcgaa ctcacacagg cgagaaacct taccactgtg actgggacgg ctgtgggtgg
4201 aaattcgccc gctccgatga actgaccagg cactaccgca aacacacagg gcaccggccc
4261 tttcagtgcc agaagtgcga cagggccttt tccaggtcgg accaccttgc cttacacatg
4321 aagaggcact ttggggatct caaaattgtc gctcctgtca aacaaactct taactttgat
4381 ttactcaaac tggctgggga tgtagaaagc aatccaggtc cactcatgaa caattcgccc
4441 ttcaccatgt ataacatgat ggagacggag ctgaagccgc cgggcccgca gcaagcttcg
4501 gggggcggcg gcggaggagg caacgccacg gcggcggcga ccggcggcaa ccagaagaac
4561 agcccggacc gcgtcaagag gcccatgaac gccttcatgg tatggtcccg ggggcagcgg
4621 cgtaagatgg cccaggagaa ccccaagatg cacaactcgg agatcagcaa gcgcctgggc
4681 gcggagtgga aacttttgtc cgagaccgag aagcggccgt tcatcgacga ggccaagcgg
4741 ctgcgcgctc tgcacatgaa ggagcacccg gattataaat accggccgcg gcggaaaacc
4801 aagacgctca tgaagaagga taagtacacg cttcccggag gcttgctggc ccccggcggg
4861 aacagcatgg cgagcggggt tggggtgggc gccggcctgg gtgcgggcgt gaaccagcgc
4921 atggacagct acgcgcacat gaacggctgg agcaacggca gctacagcat gatgcaggag
4981 cagctgggct acccgcagca cccgggcctc aacgctcacg gcgcggcaca gatgcaaccg
5041 atgcaccgct acgacgtcag cgccctgcag tacaactcca tgaccagctc gcagacctac
5101 atgaacggct cgcccaccta cagcatgtcc tactcgcagc agggcacccc cggtatggcg
5161 ctgggctcca tgggctctgt ggtcaagtcc gaggccagct ccagcccccc cgtggttacc
5221 tcttcctccc actccagggc gccctgccag gccggggacc tccgggacat gatcagcatg
5281 tacctccccg gcgccgaggt gccggagccc gctgcgccca gtagactgca catggcccag
5341 cactaccaga gcggcccggt gcccggcacg gccattaacg gcacactgcc cctgtcgcac
5401 atgtgagggc tggaagggcg aattgcctgc aggaattcac tcctcaggtg caggctgcct
5461 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
5521 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
5581 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
5641 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
5701 tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc
5761 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
5821 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
5881 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
5941 tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca
6001 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
6061 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
6121 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
6181 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
6241 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
6301 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
6361 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
6421 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
6481 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
6541 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
6601 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
6661 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
6721 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
6781 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
6841 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
6901 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
6961 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
7021 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
7081 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
7141 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
7201 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
7261 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
7321 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
7381 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
7441 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
7501 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
7561 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
7621 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
7681 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
7741 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
7801 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
7861 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
7921 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
7981 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
8041 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
8101 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
8161 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
8221 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
8281 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
8341 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
8401 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
8461 gttccgcgca catttccccg aaaagtgcca cctgg
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：BECLIN-HA WT
- 下一篇：Stat3 Flag pRc/CMV
