基本信息
| 复制子: | pUC ori |
| 质粒分类: | 病毒系列，逆病毒克隆克隆载体 |
| 原核抗性: | 氨苄青霉素Amp |
| 筛选标记: | 嘌呤霉素Puro |
| 克隆菌株: | 大肠杆菌Stbl3 |
| 培养条件: | 37℃，有氧，LB |
| 表达宿主: | 哺乳细胞 |
| 诱导方式: | 无须诱导，瞬时表达，也可稳定表达 |
| 5'测序引物: | 根据序列设计引物 |
| 3'测序引物: | 根据序列设计引物 |
质粒简介
小鼠干细胞病毒（MSCV）载体来源于小鼠胚胎干细胞病毒（MesV）和LN的逆转录病毒载体。在转染包装细胞系，pmscvpuro瞬时表达和稳定表达，或整合，成绩单包含扩展病毒包装信号Ψ+，嘌呤霉素抗性基因和目的基因。载体实现稳定、高水平的基因表达在造血干细胞和胚胎干细胞通过一个专门设计的5“长末端重复序列（LTR）。这个序列是从小鼠干细胞基因的病毒，它不同于用几个点突变其他逆转录病毒载体MoMuLV LTR和缺失。这些变化提高转录激活和阻止胚胎干细胞和胚胎癌细胞的转录YZ。作为一个结果，LTR驱动目的基因在干细胞或其他的哺乳动物细胞系的高层组成性表达。一个基因可以被克隆到的多克隆位点这LTR.小鼠磷酸甘油酸激酶（PKG）启动子下游（ppkg）的嘌呤霉素抗性基因控制表达（puror）选择抗生素在真核细胞。pmscvpuro还包含复制和大肠杆菌ampR基因在细菌繁殖和抗生素选择市局起源。质粒图谱
质粒序列
LOCUS Exported 7627 bp ds-DNA circular SYN 27-APR-2018
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..7627
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 579..920
/note="MESV Psi"
/note="packaging signal of murine embryonic stem cell
virus"
CDS 987..1403
/note="gag (truncated)"
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
promoter 1438..1937
/note="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 1958..2557
/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 2635..3187
/note="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 3199..3918
/codon_start=1
/product="enhanced GFP"
/note="EGFP"
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
LTR 4020..4534
/note="3' LTR"
/note="3' long terminal repeat from murine embryonic stem
cell virus"
protein_bind 4696..4712
/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(4720..4750)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 4765..4786
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5074..5662)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5833..6693)
/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(6694..6798)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
1381 ccctcactcc ttctctaggc gccggaatta gatctctcga ggttaacgaa ttctaccggg
1441 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca
1501 cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc
1561 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga
1621 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc
1681 tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg
1741 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg
1801 ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg
1861 gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct
1921 catctccggg cctttcgacc tgcagcccaa gcttaccatg accgagtaca agcccacggt
1981 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
2041 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
2101 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
2161 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
2221 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
2281 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
2341 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
2401 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
2461 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
2521 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg
2581 cccgaccgaa aggagcgcac gaccccatgc atccaattcc gccccccccc cctaacgtta
2641 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
2701 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
2761 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
2821 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
2881 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
2941 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
3001 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
3061 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt
3121 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
3181 gataatatgg ccacaaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
3241 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc tggcgagggc
3301 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
3361 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
3421 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
3481 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
3541 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
3601 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
3661 gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa catcgaggac
3721 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
3781 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
3841 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
3901 gacgagctgt acaagtaatg aattaattaa gaattatcaa gcttatcgat accgtcgacc
3961 tgcagccaag cttatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
4021 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
4081 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
4141 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
4201 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
4261 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
4321 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
4381 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt gtatccaata
4441 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
4501 agtgattgac tacccgtcag cgggggtctt tcatgggtaa cagtttcttg aagttggaga
4561 acaacattct gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt
4621 tgaatccaca tactccaata ctcctgaaat agttcattat ggacagcgca gaaagagctg
4681 gggagaattg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
4741 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
4801 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
4861 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
4921 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
4981 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
5041 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
5101 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
5161 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
5221 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
5281 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
5341 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
5401 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
5461 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
5521 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
5581 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
5641 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
5701 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
5761 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
5821 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
5881 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
5941 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
6001 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
6061 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
6121 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
6181 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
6241 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
6301 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
6361 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
6421 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
6481 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
6541 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
6601 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
6661 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
6721 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
6781 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
6841 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
6901 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
6961 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
7021 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
7081 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
7141 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
7201 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
7261 gtcacgacgt tgtaaaacga cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac
7321 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
7381 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
7441 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggcgatt agtccaattt
7501 gttaaagaca ggatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg
7561 aagcctatag agtacgagcc atagataaaa taaaagattt tatttagtct ccagaaaaag
7621 gggggaa
温馨提示：不可用于临床ZL。
