| 质粒名称: | pCXB-EBNA1 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 41857 |
| 存储实验室: | Shinya Yamanaka |
|
载体骨架基本信息 |
|
| 载体名称: | pCXB-EBNA1 |
| 空载体大小: | 4452 bp |
| 完整质粒大小: | 6412bp |
| 载体修饰: | human herpesvirus 4 |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | EBNA-1 |
| 插入大小: | 1926 bp |
| GenBank编号 | AJ507799 |
| 启动子: | CAG |
| 标签/融合蛋白: | EBNA-1 (a.k.a. HHV4_EBNA-1.2) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI(未破坏) |
| 3’克隆位点: | EcoRI (未破坏) |
| 5’测序引物 | pCAG-F |
| 3’测序引物 |
TTAGCCAGAAGTCAGATGCTC
|
|
文献引用方法 |
|
| 材料和方法部分: | pCXB-EBNA1 was a gift from Shinya Yamanaka (Addgene plasmid # 41857 ; http://n2t.net/addgene:41857 ; RRID:Addgene_41857) |
| 参考文献部分: | An Efficient Non-viral Method to Generate Integration-Free Human iPS Cells from Cord Blood and Peripheral Blood Cells. Okita K, Yamakawa T, Matsumura Y, Sato Y, Amano N, Watanabe A, Goshima N, Yamanaka S. Stem Cells. 2012 Nov 29. doi: 10.1002/stem.1293. 10.1002/stem.1293 PubMed 23193063 |
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 pCXB-EBNA1 6412 bp ds-DNA circular SYN 24-AUG-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCXB-EBNA1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6412)
AUTHORS Okita K, Yamakawa T, Matsumura Y, Sato Y, Amano N, Watanabe A,
Goshima N, Yamanaka S
TITLE An Efficient Non-viral Method to Generate Integration-Free Human iPS
Cells from Cord Blood and Peripheral Blood Cells.
JOURNAL Stem Cells. 2012 Nov 29. doi: 10.1002/stem.1293.
PUBMED 23193063
REFERENCE 2 (bases 1 to 6412)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 24, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6412
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(83..102)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 148..203
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(202..221)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(564..580)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(564..580)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(577..599)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 588..604
/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(612..642)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 657..678
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(803..820)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(974..1562)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1054..1073)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1733..2593)
/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 2356..2375
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2594..2698)
/gene="bla"
/label=AmpR promoter
enhancer 2729..3108
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3110..3387
/label=chicken beta-actin promoter
intron 3388..4404
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 4328..4350
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 4412..4431
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 4474..6399
/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"
ORIGIN
1 aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc
61 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga
121 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
181 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
241 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga
301 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
361 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat
421 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc
481 tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc
541 agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
601 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
661 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
721 tcgtgccagc ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
781 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
841 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
901 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
961 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
1021 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
1081 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
1141 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
1201 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
1261 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
1321 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
1381 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
1441 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
1501 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
1561 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
1621 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
1681 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
1741 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
1801 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
1861 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
1921 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
1981 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
2041 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
2101 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
2161 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
2221 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
2281 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
2341 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
2401 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
2461 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
2521 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
2581 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
2641 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
2701 catttccccg aaaagtgcca cctgggtcga cattgattat tgactagtta ttaatagtaa
2761 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
2821 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
2881 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
2941 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
3001 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
3061 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc
3121 cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta
3181 tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg gsgcgcgcca
3241 ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc
3301 aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc
3361 tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc
3421 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
3481 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg
3541 cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc
3601 ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc
3661 tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc
3721 agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag
3781 gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg
3841 tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct
3901 tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg
3961 cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag
4021 gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc
4081 cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag
4141 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc
4201 gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt
4261 ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc
4321 agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt
4381 catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat
4441 cattttggca aagaattcgc ccttgtcgac accatgtctg acgaggggcc aggtacagga
4501 cctggaaatg gcctaggaga gaagggagac acatctggac cagaaggctc cggcggcagt
4561 ggacctcaaa gaagaggggg tgataaccat ggacgaggac ggggaagagg acgaggacga
4621 ggaggcggaa gaccaggagc cccgggcggc tcaggatcag ggccaagaca tagagatggt
4681 gtccggagac cccaaaaacg tccaagttgc attggctgca aagggaccca cggtggaaca
4741 ggagcaggag caggagcggg aggggcagga gcaggagggg caggagcagg aggaggggca
4801 ggagcaggag gaggggcagg aggggcagga ggggcaggag gggcaggagc aggaggaggg
4861 gcaggagcag gaggaggggc aggaggggca ggaggggcag gagcaggagg aggggcagga
4921 gcaggaggag gggcaggagg ggcaggagca ggaggagggg caggaggggc aggaggggca
4981 ggagcaggag gaggggcagg agcaggagga ggggcaggag gggcaggagc aggaggaggg
5041 gcaggagggg caggaggggc aggagcagga ggaggggcag gagcaggagg ggcaggaggg
5101 gcaggagggg caggagcagg aggggcagga gcaggaggag gggcaggagg ggcaggaggg
5161 gcaggagcag gaggggcagg agcaggaggg gcaggagcag gaggggcagg agcaggaggg
5221 gcaggagggg caggagcagg aggggcagga ggggcaggag caggaggggc aggaggggca
5281 ggagcaggag gaggggcagg aggggcagga gcaggaggag gggcaggagg ggcaggagca
5341 ggaggggcag gaggggcagg agcaggaggg gcaggagggg caggagcagg aggggcagga
5401 ggggcaggag caggaggagg ggcaggagca ggaggggcag gagcaggagg tggaggccgg
5461 ggtcgaggag gcagtggagg ccggggtcga ggaggtagtg gaggccgggg tcgaggaggt
5521 agtggaggcc gccggggtag aggacgtgaa agagccaggg ggggaagtcg tgaaagagcc
5581 agggggagag gtcgtggacg tggagaaaag aggcccagga gtcccagtag tcagtcatca
5641 tcatccgggt ctccaccgcg caggccccct ccaggtagaa ggccattttt ccaccctgta
5701 ggggaagccg attattttga ataccaccaa gaaggtggcc cagatggtga gcctgacgtg
5761 cccccgggag cgatagagca gggccccgca gatgacccag gagaaggccc aagcactgga
5821 ccccggggtc agggtgatgg aggcaggcgc aaaaaaggag ggtggtttgg aaagcatcgt
5881 ggtcaaggag gttccaaccc gaaatttgag aacattgcag aaggtttaag agctctcctg
5941 gctaggagtc acgtagaaag gactaccgac gaaggaactt gggtcgccgg tgtgttcgta
6001 tatggaggta gtaagacctc cctttacaac ctaaggcgag gaactgccct tgctattcca
6061 caatgtcgtc ttacaccatt gagtcgtctc ccctttggaa tggcccctgg acccggccca
6121 caacctggcc cgctaaggga gtccattgtc tgttatttca tggtcttttt acaaactcat
6181 atatttgctg aggttttgaa ggatgcgatt aaggaccttg ttatgacaaa gcccgctcct
6241 acctgcaata tcagggtgac tgtgtgcagc tttgacgatg gagtagattt gcctccctgg
6301 tttccaccta tggtggaagg ggctgccgcg gagggtgatg acggagatga cggagatgaa
6361 ggaggtgatg gagatgaggg tgaggaaggg caggagtgag gatccaaggg cg
//
