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- 60908-1364
- 2025年07月11日
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- 详细信息
- 技术资料
- 保存条件：
低温运输，-20℃保存
- 保质期：
1年
- 英文名：
pCDH-EF1-MCS-T2A-Puro
- 库存：
大量
- 供应商：
信裕生物
质粒编号: 60908-1364
质粒名称: pCDH-EF1-MCS-T2A-Puro
其他名称:
质粒大小（bp）: 7087 bp
抗性: Ampicillin (氨苄青霉素)
质粒类型: 慢病毒表达载体；cDNA克隆载体
是否病毒:慢病毒（HIV)
启动子:EF-1α core
稳转或瞬转:稳表达
组成型或诱导性:组成型
表达水平:
蛋白标签:
5′ 测序引物及序列:根据序列设计引物
3′ 测序引物及序列:根据序列设计引物
GenBank号:
其他属性:用于cDNA表达和克隆；高效转染细胞，建立稳定细胞系；
单一启动子，由EF1a 启动子驱动目的基因和与抗性基因转录为双顺反子；
二者之间含一段T2A肽，能够在翻译后水平上自我剪切，保证目的基因与抗性基因翻译水平相同。
质粒图谱:
质粒序列:
LOCUS Exported 7087 bp ds-DNA circular SYN 30-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7087)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Tuesday, August 30, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..7087
/organism=”synthetic DNA construct”
/mol_type=”other DNA”
LTR 234..414
/note=”5′ LTR (truncated)”
/note=”truncated 5′ long terminal repeat (LTR) from HIV-1″
misc_feature 458..583
/note=”HIV-1 Psi”
/note=”packaging signal of human immunodeficiency virus
type 1″
misc_feature 1076..1309
/note=”RRE”
/note=”The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm.”
misc_feature 1798..1915
/note=”cPPT/CTS”
/note=”central polypurine tract and central termination
sequence of HIV-1″
promoter 1960..2171
/note=”EF-1-alpha core promoter”
/note=”core promoter for human elongation factor
EF-1-alpha”
LTR 2184..2452
/note=”5′ LTR (truncated)”
/note=”truncated 5′ long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1″
CDS 2510..2563
/codon_start=1
/product=”2A peptide from Thosea asigna virus capsid
protein”
/note=”T2A”
/note=”Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides.”
/translation=”EGRGSLLTCGDVEENPGP”
CDS 2570..3169
/codon_start=1
/gene=”pac from Streptomyces alboniger”
/product=”puromycin N-acetyltransferase”
/note=”PuroR”
/note=”confers resistance to puromycin”
/translation=”MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA”
misc_feature 3182..3770
/note=”WPRE”
/note=”woodchuck hepatitis virus posttranscriptional
regulatory element”
CDS complement(3653..3664)
/codon_start=1
/product=”Factor Xa recognition and cleavage site”
/note=”Factor Xa site”
/translation=”IEGR”
LTR 3844..4077
/note=”3′ LTR (Delta-U3)”
/note=”self-inactivating 3′ long terminal repeat (LTR) from
HIV-1″
polyA_signal 4149..4270
/note=”SV40 poly(A) signal”
/note=”SV40 polyadenylation signal”
rep_origin 4289..4424
/note=”SV40 ori”
/note=”SV40 origin of replication”
primer_bind complement(4462..4478)
/note=”M13 rev”
/note=”common sequencing primer, one of multiple similar
variants”
protein_bind 4486..4502
/bound_moiety=”lac repressor encoded by lacI”
/note=”lac operator”
/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(4510..4540)
/note=”lac promoter”
/note=”promoter for the E. coli lac operon”
protein_bind 4555..4576
/bound_moiety=”E. coli catabolite activator protein”
/note=”CAP binding site”
/note=”CAP binding activates transcription in the presence
of cAMP.”
rep_origin complement(4864..5452)
/direction=LEFT
/note=”ori”
/note=”high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication”
CDS complement(5623..6483)
/codon_start=1
/gene=”bla”
/product=”beta-lactamase”
/note=”AmpR”
/note=”confers resistance to ampicillin, carbenicillin, and
related antibiotics”
/translation=”MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW”
promoter complement(6484..6588)
/gene=”bla”
/note=”AmpR promoter”
primer_bind 7062..7078
/note=”M13 fwd”
/note=”common sequencing primer, one of multiple similar
variants”
ORIGIN
1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggttaacttt
1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
1921 tactagtaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca
1981 cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc
2041 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
2101 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
2161 ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta
2221 cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct
2281 cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc
2341 cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc
2401 ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt
2461 gaccggcgcc tactctagag ctagcgaatt cgaaggatcc gcggccgctg agggcagagg
2521 aagtcttcta acatgcggtg acgtggagga gaatcccggc ccttccggga tgaccgagta
2581 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
2641 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
2701 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
2761 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
2821 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
2881 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
2941 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
3001 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
3061 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
3121 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa tctaggtcga
3181 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
3241 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
3301 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
3361 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
3421 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
3481 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
3541 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
3601 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
3661 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
3721 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt
3781 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
3841 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt
3901 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
3961 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
4021 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
4081 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
4141 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
4201 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
4261 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca
4321 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
4381 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga
4441 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
4501 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
4561 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
4621 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
4681 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
4741 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
4801 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
4861 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
4921 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
4981 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
5041 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
5101 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
5161 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
5221 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
5281 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
5341 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
5401 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
5461 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
5521 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
5581 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
5641 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
5701 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
5761 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
5821 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
5881 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
5941 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
6001 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
6061 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
6121 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
6181 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
6241 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
6301 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
6361 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
6421 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
6481 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
6541 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
6601 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
6661 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
6721 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
6781 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
6841 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
6901 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
6961 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
7021 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
7081 caagctg
//
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pCDH-EF1-MCS-T2A-Puro
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