| 质粒名称: | pMSCV-IRES-GFP II |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 52107 |
| 存储实验室: | Dario Vignali |
|
载体骨架基本信息 |
|
| 载体名称: | pN1 |
| 载体出品公司: | Clontech |
| 载体抗性: | 氨苄西林 |
| 载体修饰: | 扩展的MCS |
| 载体类型: | 哺乳动物表达，逆转录病毒 |
| 标记/融合蛋白： | IRES-EGFP（主干网上的C终端） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 克隆方法: | 限制性酶 |
| 5' 测序引物 | pLXSN_5_prime |
|
文献引用方法 |
|
| 材料和方法部分: | pMSCV-IRES-GFP II (pMIG II) was a gift from Dario Vignali (Addgene plasmid # 52107 ; http://n2t.net/addgene:52107 ; RRID:Addgene_52107) |
| 参考文献部分: | Generation of T-cell receptor retrogenic mice. Holst J, Szymczak-Workman AL, Vignali KM, Burton AR, Workman CJ, Vignali DA. Nat Protoc. 2006;1(1):406-17. 10.1038/nprot.2006.61 PubMed 17406263 |
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 pMSCV-IRES-GFP II 6512 bp ds-DNA circular SYN 12-NOV-2018
DEFINITION a bicistronic IRES-GFP containing retroviral vector, MCS is expanded
from pMSCV-IRES-GFP.
ACCESSION .
VERSION .
KEYWORDS pMSCV-IRES-GFP II (pMIG II)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6512)
AUTHORS Holst J, Szymczak-Workman AL, Vignali KM, Burton AR, Workman CJ,
Vignali DA
TITLE Generation of T-cell receptor retrogenic mice.
JOURNAL Nat Protoc. 2006;1(1):406-17.
PUBMED 17406263
REFERENCE 2 (bases 1 to 6512)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 12, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6512
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..517
/label=5' LTR
/note="5' long terminal repeat from murine embryonic stem
cell virus"
misc_feature 581..922
/label=MESV Psi
/note="packaging signal of murine embryonic stem cell
virus"
CDS 989..1405
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1335..1357
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1373..1389
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
primer_bind complement(1618..1635)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 1845..1864
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 2026..2745
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(2071..2092)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2332..2351)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2679..2700
/label=EGFP-C
/note="EGFP, forward primer"
LTR 2907..3421
/label=3' LTR
/note="3' long terminal repeat from murine embryonic stem
cell virus"
protein_bind 3583..3599
/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(3607..3637)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3652..3673
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3790..3807)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3961..4549)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4041..4060)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4720..5580)
/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 5343..5362
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5581..5685)
/gene="bla"
/label=AmpR promoter
primer_bind 5753..5771
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(5809..5831)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 5931..5950
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6144..6166
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 6294..6313
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
1381 agccctcact ccttctctag gcgccggaat tcagatctta cgtagctagc ggatcccaat
1441 tgctcgagcg ggatcaattc cgcccccccc ctaacgttac tggccgaagc cgcttggaat
1501 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
1561 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
1621 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
1681 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
1741 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
1801 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
1861 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
1921 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
1981 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgata ataccatggt gagcaagggc
2041 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
2101 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
2161 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
2221 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
2281 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
2341 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
2401 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
2461 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
2521 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
2581 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
2641 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
2701 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct
2761 agagtcgacc tgcaggcatg caagcttcag gtagccggct aacgttaaca accggtacct
2821 ctagaactat agctagcatg cgcaaattta aagcgctgat atcgataaaa taaaagattt
2881 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
2941 cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag
3001 atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc
3061 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag
3121 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
3181 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
3241 cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc
3301 gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct
3361 cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca
3421 tgggtaacag tttcttgaag ttggagaaca acattctgag ggtaggagtc gaatattaag
3481 taatcctgac tcaattagcc actgttttga atccacatac tccaatactc ctgaaatagt
3541 tcattatgga cagcgcagaa agagctgggg agaattgtga aattgttatc cgctcacaat
3601 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
3661 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
3721 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
3781 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
3841 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
3901 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
3961 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
4021 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
4081 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
4141 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
4201 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
4261 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
4321 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
4381 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
4441 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
4501 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
4561 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
4621 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
4681 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
4741 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
4801 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
4861 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
4921 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
4981 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
5041 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
5101 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
5161 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
5221 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
5281 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
5341 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
5401 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
5461 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
5521 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
5581 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
5641 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
5701 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
5761 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
5821 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
5881 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
5941 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
6001 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
6061 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
6121 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat
6181 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
6241 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
6301 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
6361 ccggcgtaga ggcgattagt ccaatttgtt aaagacagga tatcagtggt ccaggctcta
6421 gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa
6481 aagattttat ttagtctcca gaaaaagggg gg
//
