| 质粒名称: | PL-SIN-EOS-C(3+)-EiP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 21313 |
| 存储实验室: | James Ellis |
|
载体骨架基本信息 |
|
| 载体名称: | PL-SIN lentiviral vector |
| 载体出品公司: | Buzina et al., PLoS Genetics, 2008 |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 6141 bp |
| 完整质粒大小: | 8088 bp |
| 载体类型: | 哺乳细胞表达，Lentiviral |
| 筛选标记: | 嘌呤霉素 |
| 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: | Enhanced Green Fluorescence Protein, Puromycin resistance gene |
| 插入大小: | 1947 bp |
| 基因又名: | EGFP-IRES-Puro |
| 启动子 | EOS-C(3+) promoter region |
| 克隆方法: | 未知 |
| 5' 测序引物 | n/a |
| 3' 测序引物 | n/a |
|
文献引用方法 |
|
| 材料和方法部分: | PL-SIN-EOS-C(3+)-EiP was a gift from James Ellis (爱迪基因 plasmid # 21313) |
| 参考文献部分: | Isolation of human iPS cells using EOS lentiviral vectors to select for pluripotency. Hotta A, Cheung AY, Farra N, Vijayaragavan K, Seguin CA, Draper JS, Pasceri P, Maksakova IA, Mager DL, Rossant J, Bhatia M, Ellis J. Nat Methods. 2009 May . 6(5):370-6. 10.1038/nmeth.1325 PubMed 19404254 |
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
EiP质粒图谱.png)
载体简介
载体序列
LOCUS PL-SIN-EOS-C(3+)-EiP 8482 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Lentiviral EOS reporter with Oct3/4 enhancer (x3), expresses EGFP
and Puro resistance.
ACCESSION .
VERSION .
KEYWORDS PL-SIN-EOS-C(3+)-EiP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8482)
AUTHORS Hotta A, Cheung AY, Farra N, Vijayaragavan K, Seguin CA, Draper JS,
Pasceri P, Maksakova IA, Mager DL, Rossant J, Bhatia M, Ellis J
TITLE Isolation of human iPS cells using EOS lentiviral vectors to select
for pluripotency.
JOURNAL Nat Methods. 2009 May . 6(5):370-6.
PUBMED 19404254
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|PL-SIN-EOS-C(3+)-EiP|
FEATURES Location/Qualifiers
source 1..8482
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(29..51)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 151..170
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 364..386
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 378..395
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
LTR 785..1042
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1693..1810
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
primer_bind 2762..2778
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
regulatory 3675..3684
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3681..4400
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3726..3747)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3987..4006)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4334..4355
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 4441..5027
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(4608..4625)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 4835..4854
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 5028..5627
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(5028..5047)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 5524..5544
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 5769..6002
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(6039..6061)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6050..6066
/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(6074..6104)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6119..6140
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6257..6274)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6428..7016)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6508..6527)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7809..7828
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8047..8151)
/gene="bla"
/label=AmpR promoter
promoter complement(8188..8384)
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 8202..8337
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(8256..8275)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 8455..8473
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gcggccgctg gaagggctaa
421 tttggtccca aaaaagacaa gagatccttg atctgtggat ctaccacaca caaggctact
481 tccctgattg gcagaactac acaccagggc cagggatcag atatccactg acctttggat
541 ggtgcttcaa gttagtacca gttgaaccag agcaagtaga agaggccaaa taaggagaga
601 agaacagctt gttacaccct atgagccagc atgggatgga ggacccggag ggagaagtat
661 tagtgtggaa gtttgacagc ctcctagcat ttcgtcacat ggcccgagag ctgcatccgg
721 agtactacaa agactgctga catcgagctt tctacaaggg actttccgct ggggactttc
781 cagggaggtg tggcctgggc gggactgggg agtggcgagc cctcagatgc tacatataag
841 cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
901 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgctcaaa
961 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
1021 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagtaaagc
1081 cagaggagat ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
1141 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
1201 ggtgcgagag cgtcggtatt aagcggggga gaattagata aatgggaaaa aattcggtta
1261 aggccagggg gaaagaaaca atataaacta aaacatatag tatgggcaag cagggagcta
1321 gaacgattcg cagttaatcc tggcctttta gagacatcag aaggctgtag acaaatactg
1381 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
1441 atagcagtcc tctattgtgt gcatcaaagg atagatgtaa aagacaccaa ggaagcctta
1501 gataagatag aggaagagca aaacaaaagt aagaaaaagg cacagcaagc agcagctgac
1561 acaggaaaca acagccaggt cagccaaaat taccctatag tgcagaacct ccaggggcaa
1621 atggtacatc aggccatatc acctagaact ttaaattaag acagcagtac aaatggcagt
1681 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
1741 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
1801 tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggaaag gaccagcaaa
1861 gctcctctgg aaaggtgaag gggcagtagt aatacaagat aatagtgaca taaaagtagt
1921 gccaagaaga aaagcaaaga tcatcaggga ttatggaaaa cagatggcag gtgatgattg
1981 tgtggcaagt agacaggatg aggattaaca catggaaaag attagtaaaa caccatagct
2041 ctagagcgat ccgatcttca gacctggagg aggcgatatg agggacaatt ggagaagtga
2101 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
2161 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
2221 cttgggagca gcaggaagca ctatgggctg cacgtcaatg acgctgacgg tacaggccag
2281 acaattattg tctgatatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
2341 acagcatctg ttgcaactca cagtctgggg catcaaacag ctccaggcaa gaatcctggc
2401 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
2461 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
2521 ttggaataac atgacctgga tggagtggga cagagaaatt aacaattaca caagcttggt
2581 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
2641 accattatcg tttcagaccc acctcccaat cccgagggga cccgacaggc ccgaaggaat
2701 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcctc
2761 cctttatcca gccctcactc cttctctagg cgccggaatt cgggtgtggg gaggttgtag
2821 cccgaccctg cccctccccc cagggaggtt gagagttctg ggcagacggc agatgcataa
2881 caaaggtgca tgatagctct gccctggggg cagagaagat ggttggggag gggtccctct
2941 cgtcctagct cgagcttaat cactagtgaa ttcgggtgtg gggaggttgt agcccgaccc
3001 tgcccctccc cccagggagg ttgagagttc tgggcagacg gcagatgcat aacaaaggtg
3061 catgatagct ctgccctggg ggcagagaag atggttgggg aggggtccct ctcgtcctag
3121 ctcgagctta atcactagtg aattcgggtg tggggaggtt gtagcccgac cctgcccctc
3181 cccccaggga ggttgagagt tctgggcaga cggcagatgc ataacaaagg tgcatgatag
3241 ctctgccctg ggggcagaga agatggttgg ggaggggtcc ctctcgtcct agctcgagct
3301 taatcactag tgaattctcc cctcccccag cctgaaacct gcttgctcag gggtggagct
3361 tcctgctcat tcgttctgcc acgcccactg ctggaacctg cggagccaca cccgtgcacc
3421 tttctactgg accagagatt attcggcggg aatcgggtcc cctccccctt ccttcataac
3481 tagtgtcgca actataaaat ttgagccttg atcagagtaa ctgtcttggc tacattcttt
3541 cttccgcccc gtctagattc ctctcttaaa gctcgagcgg ccttctcagt cgaaccgttc
3601 acgttgcgag ctgctggcgg ccgcaacaag atctgcgatc taagtaagct tggcattccg
3661 gtactgttgg taaagccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
3721 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
3781 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
3841 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
3901 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
3961 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
4021 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
4081 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
4141 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
4201 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
4261 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
4321 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
4381 tggacgagct gtacaagtaa agcggccaac ctcgacgtta ttcccttcga tcgaattccg
4441 cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
4501 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
4561 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
4621 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
4681 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
4741 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
4801 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
4861 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
4921 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
4981 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg accgagtaca
5041 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
5101 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
5161 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
5221 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
5281 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
5341 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
5401 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
5461 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
5521 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
5581 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg
5641 acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atcgatttcg aacccggggt
5701 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
5761 ggggggactg gaagggctaa ttcactccca aagaagacaa gatctgcttt ttgcctgtac
5821 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
5881 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
5941 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
6001 cagaattcta gcttggcgta atcatggtca tagctgttcc tgtgtgaaat tgttatccgc
6061 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
6121 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
6181 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
6241 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
6301 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
6361 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
6421 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
6481 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
6541 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
6601 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
6661 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
6721 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
6781 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
6841 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
6901 cacttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
6961 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
7021 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
7081 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
7141 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
7201 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
7261 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
7321 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
7381 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
7441 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gtgccattgc
7501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
7561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
7621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
7681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
7741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
7801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
7861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
7921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
7981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
8041 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
8101 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
8161 ccgaaaagtg ccacctggga ctagcttttt gcaaaagcct aggcctccaa aaaagcctcc
8221 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
8281 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
8341 cggagttagg ggcgggacta tggttgctga ctaattgaga tgtagcttgc atgccgacat
8401 ggattattga ctagtcccta agaaaccatt attatcatga cattaaccta taaaaatagg
8461 cgtatcacga ggccctttcg tc
//
