| 质粒名称: | pCXWB-EBNA1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 37624 |
| 存储实验室: | Shinya Yamanaka |
|
载体骨架基本信息 |
|
| 载体名称: | pCXWB |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 5078 bp |
| 完整质粒大小: | 7004 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: | EBNA-1 |
| 物种来源: | human herpesvirus 4 |
| 插入大小: | 1926 bp |
| 基因又名: | HHV4-EBNA-1.1 |
| GenBank编号 | 3783709 |
| 启动子 | CAG |
| 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 | WPRE-R |
|
文献引用方法 |
|
| 材料和方法部分: | pCXWB-EBNA1 was a gift from Shinya Yamanaka (爱迪基因 plasmid # 37624) |
| 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 pCXWB-EBNA1 7037 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Non-replicating episomal expression of EBNA1, which enhances
transfection efficiency and expression from episomal plasmids.
ACCESSION .
VERSION .
KEYWORDS pCXWB-EBNA1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7037)
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
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pCXWB-EBNA1|
FEATURES Location/Qualifiers
source 1..7037
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 3..382
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 384..660
/label=chicken beta-actin promoter
intron 661..1677
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1601..1623
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1685..1704
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 1747..3672
/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"
misc_feature 3716..4304
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3769..3789)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4187..4198)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(4394..4413)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 4459..4514
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(4513..4532)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(4875..4891)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4875..4891)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4888..4910)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4899..4915
/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(4923..4953)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4968..4989
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5114..5131)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5285..5873)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5365..5384)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6044..6904)
/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 6667..6686
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6905..7009)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
61 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
121 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
181 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
241 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
301 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
361 attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat
421 ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc
481 gatgggggcg gggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
541 gggcgggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
1261 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt cgcccttgtc
1741 gacaccatgt ctgacgaggg gccaggtaca ggacctggaa atggcctagg agagaaggga
1801 gacacatctg gaccagaagg ctccggcggc agtggacctc aaagaagagg gggtgataac
1861 catggacgag gacggggaag aggacgagga cgaggaggcg gaagaccagg agccccgggc
1921 ggctcaggat cagggccaag acatagagat ggtgtccgga gaccccaaaa acgtccaagt
1981 tgcattggct gcaaagggac ccacggtgga acaggagcag gagcaggagc gggaggggca
2041 ggagcaggag gggcaggagc aggaggaggg gcaggagcag gaggaggggc aggaggggca
2101 ggaggggcag gaggggcagg agcaggagga ggggcaggag caggaggagg ggcaggaggg
2161 gcaggagggg caggagcagg aggaggggca ggagcaggag gaggggcagg aggggcagga
2221 gcaggaggag gggcaggagg ggcaggaggg gcaggagcag gaggaggggc aggagcagga
2281 ggaggggcag gaggggcagg agcaggagga ggggcaggag gggcaggagg ggcaggagca
2341 ggaggagggg caggagcagg aggggcagga ggggcaggag gggcaggagc aggaggggca
2401 ggagcaggag gaggggcagg aggggcagga ggggcaggag caggaggggc aggagcagga
2461 ggggcaggag caggaggggc aggagcagga ggggcaggag gggcaggagc aggaggggca
2521 ggaggggcag gagcaggagg ggcaggaggg gcaggagcag gaggaggggc aggaggggca
2581 ggagcaggag gaggggcagg aggggcagga gcaggagggg caggaggggc aggagcagga
2641 ggggcaggag gggcaggagc aggaggggca ggaggggcag gagcaggagg aggggcagga
2701 gcaggagggg caggagcagg aggtggaggc cggggtcgag gaggcagtgg aggccggggt
2761 cgaggaggta gtggaggccg gggtcgagga ggtagtggag gccgccgggg tagaggacgt
2821 gaaagagcca gggggggaag tcgtgaaaga gccaggggga gaggtcgtgg acgtggagaa
2881 aagaggccca ggagtcccag tagtcagtca tcatcatccg ggtctccacc gcgcaggccc
2941 cctccaggta gaaggccatt tttccaccct gtaggggaag ccgattattt tgaataccac
3001 caagaaggtg gcccagatgg tgagcctgac gtgcccccgg gagcgataga gcagggcccc
3061 gcagatgacc caggagaagg cccaagcact ggaccccggg gtcagggtga tggaggcagg
3121 cgcaaaaaag gagggtggtt tggaaagcat cgtggtcaag gaggttccaa cccgaaattt
3181 gagaacattg cagaaggttt aagagctctc ctggctagga gtcacgtaga aaggactacc
3241 gacgaaggaa cttgggtcgc cggtgtgttc gtatatggag gtagtaagac ctccctttac
3301 aacctaaggc gaggaactgc ccttgctatt ccacaatgtc gtcttacacc attgagtcgt
3361 ctcccctttg gaatggcccc tggacccggc ccacaacctg gcccgctaag ggagtccatt
3421 gtctgttatt tcatggtctt tttacaaact catatatttg ctgaggtttt gaaggatgcg
3481 attaaggacc ttgttatgac aaagcccgct cctacctgca atatcagggt gactgtgtgc
3541 agctttgacg atggagtaga tttgcctccc tggtttccac ctatggtgga aggggctgcc
3601 gcggagggtg atgacggaga tgacggagat gaaggaggtg atggagatga gggtgaggaa
3661 gggcaggagt gaggatccaa gggcgaattc aagcttcggg gactagtcat atgataatca
3721 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
3781 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
3841 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
3901 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
3961 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
4021 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
4081 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
4141 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
4201 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
4261 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcggt aaattcactc
4321 ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca
4381 aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
4441 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
4501 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
4561 agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt
4621 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata
4681 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt
4741 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac
4801 tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg cagcccaagc
4861 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
4921 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
4981 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
5041 cggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
5101 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
5161 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
5221 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
5281 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
5341 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
5401 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
5461 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
5521 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
5581 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
5641 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
5701 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
5761 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
5821 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
5881 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
5941 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
6001 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
6061 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
6121 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
6181 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
6241 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
6301 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
6361 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
6421 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
6481 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
6541 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
6601 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
6661 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
6721 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
6781 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
6841 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
6901 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
6961 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
7021 gaaaagtgcc acctggg
//
