| 质粒名称: | MSGV Hu Acceptor PGK-NGFR |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 64270 |
| 存储实验室: | David Ott |
|
载体骨架基本信息 |
|
| 载体名称: | pMSGV |
| 载体出品公司: | Richard Morgan NCI |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 8070 bp |
| 完整质粒大小: | -- |
| 载体修饰: | TCR克隆插入：PGK启动子驱动的tcrv区基因受体/恒定区基因tNGFR |
| 载体类型: | 逆转录病毒 |
| 筛选标记: | tNGFR |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 30 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | MSGV Hu Acceptor PGK-NGFR was a gift from David Ott (Addgene plasmid # 64270 ; http://n2t.net/addgene:64270 ; RRID:Addgene_64270) |
| 参考文献部分: | Production of retroviral constructs for effective transfer and expression of T-cell receptor genes using Golden Gate cloning. Coren LV, Jain S, Trivett MT, Ohlen C, Ott DE. Biotechniques. 2015 Mar 1;58(3):135-9. doi: 10.2144/000114265. eCollection 2015. 10.2144/000114265 PubMed 25757546 |
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 MSGV Hu Acceptor PGK-NGFR 8069 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Retroviral golden gate cloning vector for expression of functional
Human T-Cell Receptor genes with NGFR.
ACCESSION .
VERSION .
KEYWORDS MSGV Hu Acceptor PGK-NGFR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8069)
AUTHORS Coren LV, Jain S, Trivett MT, Ohlen C, Ott DE
TITLE Production of retroviral constructs for effective transfer and
expression of T-cell receptor genes using Golden Gate cloning.
JOURNAL Biotechniques. 2015 Mar 1;58(3):135-9. doi: 10.2144/000114265.
eCollection 2015.
PUBMED 25757546
REFERENCE 2 (bases 1 to 8069)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8069
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..518
/label=5' LTR
/note="5' long terminal repeat from murine embryonic stem
cell virus"
primer_bind 1358..1380
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1396..1412
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
primer_bind 1750..1766
/label=pBMN 5'
/note="MMLV sequence, for inserts in pBMN retroviral
vector"
promoter 2911..3409
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(2946..2969)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 3342..3361
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
misc_feature 4290..4342
/label=MCS
/note="multiple cloning site"
LTR 4386..4900
/label=3' LTR
/note="3' long terminal repeat from murine embryonic stem
cell virus"
protein_bind 5072..5088
/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(5096..5126)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5141..5162
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5279..5296)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5450..6038)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5530..5549)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6209..7069)
/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 6832..6851
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7070..7174)
/gene="bla"
/label=AmpR promoter
primer_bind 7242..7260
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(7298..7320)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 7420..7439
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7633..7655
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 7647..7664
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 7648..7664
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7863..7882
/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 ctgaaaatga ccctgtgcct tatttgaact aaccaatcag
301 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
361 cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtattccca
421 ataaagcctc ttgctgtttg catccgaatc gtggactcgc tgatccttgg gagggtctcc
481 tcagattgat tgactgccca cctcgggggt ctttcatttg gaggttccac cgagatttgg
541 agacccctgc ctagggacca ccgacccccc cgccgggagg taagctggcc agcggtcgtt
601 tcgtgtctgt ctctgtcttt gtgcgtgttt gtgccggcat ctaatgtttg cgcctgcgtc
661 tgtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttcg
721 gaacacccgg ccgcaaccct gggagacgtc ccagggactt cgggggccgt ttttgtggcc
781 cgacctgagt ccaaaaatcc cgatcgtttt ggactctttg gtgcaccccc cttagaggag
841 ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt
901 ttgctttcgg tttgggaccg aagccgcgcc gcgcgtcttg tctgctgcag catcgttctg
961 tgttgtctct gtctgactgt gtttctgtat ttgtctgaga atatgggccc gggctagcct
1021 gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac
1081 aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca
1141 acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt
1201 aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg
1261 acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag
1321 cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc
1381 ccgcctcgat cctcccttta tccagccctc actccttctc taggcgcccc catatggcca
1441 tatgagatct tatatggggc acccccgccc cttgtaaact tccctgaccc tgacatgaca
1501 agagttacta acagcccctc tctccaagct cacttacagg ctctctactt agtccagcac
1561 gaagtctgga gacctctggc ggcagcctac caagaacaac tggaccgacc ggtggtacct
1621 cacccttacc gagtcggcga cacagtgtgg gtccgccgac accagactaa gaacctagaa
1681 cctcgctgga aaggacctta cacagtcctg ctgaccaccc ccaccgccct caaagtagac
1741 ggcatcgcag cttggataca cgccgcccac gtgaaggctg ccgaccccgg gggtggacca
1801 tcctctagac cgcaacgtta attaagccac caagtcttct ttttttgaag acttcagaac
1861 cctgatcctg ccgtgtacca gctgcgggac agcaagagca gcgacaagag cgtgtgcctg
1921 ttcaccgact tcgacagcca gaccaacgtg tcccagagca aggacagcga cgtgtacatc
1981 accgacaaga ccgtgctgga catgcggagc atggacttca agagcaacag cgccgtggcc
2041 tggtccaaca agagcgattt cgcctgcgcc aacgccttca acaacagcat tatccccgag
2101 gacacattct tcccaagccc cgagagcagc tgcgacgtga agctggtgga aaagagcttc
2161 gagacagaca ccaacctgaa cttccagaac ctgagcgtga tcggcttccg gatcctgctg
2221 ctgaaggtgg ccggcttcaa cctgctgatg accctgagac tgtggtccag ccgggccaag
2281 agatctggca gcggcgccac caatttcagc ctgctgaaac aggccggcga cgtggaagag
2341 aaccctaagt cttctttttg aagacttgat ctgaacaagg tgttcccccc agaggtggcc
2401 gtgttcgagc cttctgaggc cgagatcagc cacacccaga aagccaccct cgtgtgcctg
2461 gccaccggct ttttccccga ccacgtggaa ctgtcttggt gggtcaacgg caaagaggtg
2521 cacagcggcg tgtccaccga tccccagcct ctgaaagaac agcccgccct gaacgacagc
2581 cggtactgcc tgagcagcag actgagagtg tccgccacct tctggcagaa cccccggaac
2641 cacttcagat gccaggtgca gttctacggc ctgagcgaga acgacgagtg gacccaggac
2701 agagccaagc ccgtgaccca gatcgtgtct gccgaagcct ggggcagagc cgattgcggc
2761 tttaccagcg tgtcctatca gcagggcgtg ctgagcgcca ccatcctgta cgagatcctg
2821 ctgggcaagg ccaccctgta cgccgtgctg gtgtctgccc tggtgctgat ggccatggtc
2881 aagcggaagg acttctgaat ttaaattacc gggtagggga ggcgcttttc ccaaggcagt
2941 ctggagcatg cgctttagca gccccgctgg cacttggcgc tacacaagtg gcctctggcc
3001 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt
3061 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc
3121 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac
3181 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt
3241 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct
3301 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa
3361 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcaggcg
3421 gccgcaccat gggggcaggt gccactggcc gcgccatgga cgggccgcgc ctgctgctgt
3481 tgctgcttct gggggtgtcc cttggaggtg ccaaggaggc atgccccaca ggcctgtaca
3541 cacacagcgg tgagtgctgc aaagcctgca acctgggcga gggtgtggcc cagccttgtg
3601 gagccaacca gaccgtgtgt gagccctgcc tggacagcgt gacgttctcc gacgtggtga
3661 gcgcgaccga gccgtgcaag ccgtgcaccg agtgcgtggg gctccagagc atgtcggcgc
3721 catgcgtgga ggccgacgac gccgtgtgcc gctgcgccta cggctactac caggatgaga
3781 cgactgggcg ctgcgaggcg tgccgcgtgt gcgaggcggg ctcgggcctc gtgttctcct
3841 gccaggacaa gcagaacacc gtgtgcgagg agtgccccga cggcacgtat tccgacgagg
3901 ccaaccacgt ggacccgtgc ctgccctgca ccgtgtgcga ggacaccgag cgccagctcc
3961 gcgagtgcac acgctgggcc gacgccgagt gcgaggagat ccctggccgt tggattacac
4021 ggtccacacc cccagagggc tcggacagca cagcccccag cacccaggag cctgaggcac
4081 ctccagaaca agacctcata gccagcacgg tggcaggtgt ggtgaccaca gtgatgggca
4141 gctcccagcc cgtggtgacc cgaggcacca ccgacaacct catccctgtc tattgctcca
4201 tcctggctgc tgtggttgtg ggtcttgtgg cctacatagc cttcaagagg tggaacagct
4261 gagcggccgc tacgtagatc gacggatccc tcgagctcaa gcttcgaatt ctgcagtcga
4321 cggtaccgcg ggcccgggat ccgataaaat aaaagatttt atttagtctc cagaaaaagg
4381 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
4441 aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag
4501 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
4561 ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag
4621 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
4681 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
4741 caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtgtat
4801 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
4861 cctctgagtg attgactacc cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt
4921 tggagaacaa cattctgagg gtaggagtcg aatattaagt aatcctgact caattagcca
4981 ctgttttgaa tccacatact ccaatactcc tgaaatccat cgatggagtt cattatggac
5041 agcgcagaaa gagctgggga gaattgtgaa attgttatcc gctcacaatt ccacacaaca
5101 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
5161 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
5221 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
5281 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
5341 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
5401 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
5461 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
5521 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
5581 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
5641 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
5701 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
5761 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
5821 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
5881 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
5941 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
6001 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
6061 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
6121 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
6181 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
6241 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
6301 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
6361 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
6421 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
6481 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
6541 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
6601 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
6661 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
6721 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
6781 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
6841 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
6901 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
6961 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
7021 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
7081 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
7141 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
7201 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
7261 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
7321 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
7381 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
7441 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
7501 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
7561 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
7621 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccacg ctctccctta
7681 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
7741 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
7801 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
7861 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
7921 acgatgcgtc cggcgtagag gcgatttaaa gacaggatat cagtggtcca ggctctagtt
7981 ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag
8041 attttattta gtctccagaa aaagggggg
//
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