| 质粒名称: | murine TCR OTI-2A.pMIG II |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 52111 |
| 存储实验室: | Dario Vignali |
|
载体骨架基本信息 |
|
| 载体名称: | pMIG II |
| 载体出品公司: | Dario Vignali, Addgene plasmid 52107 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 8155 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，逆转录病毒 |
| 筛选标记: | -- |
| 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: | T cell receptor alpha chain,Valpha2 |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | T2A（来自Thosea asigna病毒）（插入物上的C末端） |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | pLXSN5 |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | murine TCR OTI-2A.pMIG II was a gift from Dario Vignali (Addgene plasmid # 52111 ; http://n2t.net/addgene:52111 ; RRID:Addgene_52111) |
| 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 murine TCR OTI-2A.pMIG II 8327 bp ds-DNA circular SYN 16-APR-2019
DEFINITION retroviral expression of both TCRalpha and TCRbeta using a single
vector.
ACCESSION .
VERSION .
KEYWORDS murine TCR OTI-2A.pMIG II
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8327)
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 8327)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 16, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8327
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 579..920
/label=MESV Psi
/note="packaging signal of murine embryonic stem cell
virus"
CDS 987..1403
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1333..1355
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1371..1387
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
CDS 2248..2301
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
primer_bind complement(3431..3448)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 3658..3677
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 3839..4558
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3884..3905)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4145..4164)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4492..4513
/label=EGFP-C
/note="EGFP, forward primer"
LTR 4720..5234
/label=3' LTR
/note="3' long terminal repeat from murine embryonic stem
cell virus"
protein_bind 5396..5412
/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(5420..5450)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5465..5486
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5603..5620)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5774..6362)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5854..5873)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6533..7393)
/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 7156..7175
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7394..7498)
/gene="bla"
/label=AmpR promoter
primer_bind 7566..7584
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(7622..7644)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 7744..7763
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7957..7979
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 8107..8126
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
LTR 8326..515
/label=5' LTR
/note="5' long terminal repeat from murine embryonic stem
cell virus"
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 gccggaattc agatctacca tggacaagat cctgacagca
1441 tcgtttttac tcctaggcct tcacctagct ggggtgaatg gccagcagca ggagaaacgt
1501 gaccagcagc aggtgagaca aagtccccaa tctctgacag tctgggaagg agagaccgca
1561 attctgaact gcagttatga ggacagcact tttaactact tcccatggta ccagcagttc
1621 cctggggaag gccctgcact cctgatatcc atacgttcag tgtccgataa aaaggaagat
1681 ggacgattca caatcttctt caataaaagg gagaaaaagc tctccttgca catcacagac
1741 tctcagcctg gagactcagc tacctacttc tgtgcagcaa gtgacaacta tcagttgatc
1801 tggggctctg ggaccaagct aattataaag ccagacatcc agaacccaga acctgctgtg
1861 taccagttaa aagatcctcg gtctcaggac agcaccctct gcctgttcac cgactttgac
1921 tcccaaatca atgtgccgaa aaccatggaa tctggaacgt tcatcactga caaaactgtg
1981 ctggacatga aagctatgga ttccaagagc aatggggcca ttgcctggag caaccagaca
2041 agcttcacct gccaagatat cttcaaagag accaacgcca cctaccccag ttcagacgtt
2101 ccctgtgatg ccacgttgac tgagaaaagc tttgaaacag atatgaacct aaactttcaa
2161 aacctgtcag ttatgggact ccgaatcctc ctgctgaaag tagccggatt taacctgctc
2221 atgacgctga ggctgtggtc ctccggagag ggcagaggaa gtctgctaac atgcggtgac
2281 gtcgaggaga atcctggccc aatgtctaac actgtcctcg ctgattctgc ctggggcatc
2341 accctgctat cttgggttac tgtctttctc ttgggaacaa gttcagcaga ttctggggtt
2401 gtccagtctc caagacacat aatcaaagaa aagggaggaa ggtccgttct gacgtgtatt
2461 cccatctctg gacatagcaa tgtggtctgg taccagcaga ctctggggaa ggaattaaag
2521 ttccttattc agcattatga aaaggtggag agagacaaag gattcctacc cagcagattc
2581 tcagtccaac agtttgatga ctatcactct gaaatgaaca tgagtgcctt ggaactggag
2641 gactctgcta tgtacttctg tgccagctct cgggccaatt atgaacagta cttcggtccc
2701 ggcaccaggc tcacggtttt agaggatctg agaaatgtga ctccacccaa ggtctccttg
2761 tttgagccat caaaagcaga gattgcaaac aaacaaaagg ctaccctcgt gtgcttggcc
2821 aggggcttct tccctgacca cgtggagctg agctggtggg tgaatggcaa ggaggtccac
2881 agtggggtca gcacggaccc tcaggcctac aaggagagca attatagcta ctgcctgagc
2941 agccgcctga gggtctctgc taccttctgg cacaatcctc gaaaccactt ccgctgccaa
3001 gtgcagttcc atgggctttc agaggaggac aagtggccag agggctcacc caaacctgtc
3061 acacagaaca tcagtgcaga ggcctggggc cgagcagact gtggaatcac ttcagcatcc
3121 tatcatcagg gggttctgtc tgcaaccatc ctctatgaga tcctactggg gaaggccacc
3181 ctatatgctg tgctggtcag tggcctggtg ctgatggcca tggtcaagaa aaaaaattcc
3241 tgaggatccc aattgctcga gcgggatcaa ttccgccccc cccctaacgt tactggccga
3301 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
3361 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
3421 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
3481 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
3541 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
3601 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
3661 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
3721 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
3781 cgtctaggcc ccccgaatca cggggacgtg gttttccttt gaaaaacacg ataataccat
3841 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
3901 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
3961 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
4021 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
4081 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
4141 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
4201 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
4261 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
4321 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
4381 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
4441 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
4501 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag
4561 cggccgcgac tctagagtcg acctgcaggc atgcaagctt caggtagccg gctaacgtta
4621 acaaccggta cctctagaac tatagctagc atgcgcaaat ttaaagcgct gatatcgata
4681 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
4741 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa
4801 tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat
4861 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
4921 gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
4981 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
5041 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
5101 tccgatagac tgcgtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
5161 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
5221 cgggggtctt tcatgggtaa cagtttcttg aagttggaga acaacattct gagggtagga
5281 gtcgaatatt aagtaatcct gactcaatta gccactgttt tgaatccaca tactccaata
5341 ctcctgaaat agttcattat ggacagcgca gaaagagctg gggagaattg tgaaattgtt
5401 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
5461 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
5521 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
5581 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
5641 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
5701 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
5761 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
5821 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
5881 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
5941 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
6001 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
6061 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
6121 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
6181 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
6241 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
6301 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
6361 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
6421 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
6481 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
6541 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
6601 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
6661 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
6721 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
6781 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
6841 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
6901 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
6961 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
7021 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
7081 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
7141 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
7201 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
7261 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
7321 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
7381 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
7441 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
7501 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
7561 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
7621 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
7681 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
7741 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
7801 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
7861 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
7921 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
7981 cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
8041 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
8101 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
8161 ggccacgatg cgtccggcgt agaggcgatt agtccaattt gttaaagaca ggatatcagt
8221 ggtccaggct ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc
8281 atagataaaa taaaagattt tatttagtct ccagaaaaag gggggaa
//
