| 质粒名称: | pMD2-VSVGmut |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 182229 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pMD2 |
| 空载体大小: | 4284 bp |
| 完整质粒大小: | 5820 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | VSVGmut/G glycoprotein |
| 物种来源: | Vesicular Stomatitis Virus |
| 插入大小: | 1536 bp |
| 基因突变: | K47Q, R354A |
| 启动子: | CMV |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | tgtgtgctggcccatcactttggcaaagcacgtgagatct |
| 3’测序引物 | tgtgtgctggcccatcactttggcaaagcacgtgagatct |
|
文献引用方法 |
|
| 材料和方法部分: | pMD2-VSVGmut was a gift from Michael Birnbaum (Addgene plasmid # 182229 ; http://n2t.net/addgene:182229 ; RRID:Addgene_182229) |
| 参考文献部分: | Antigen identification and high-throughput interaction mapping by reprogramming viral entry. Dobson CS, Reich AN, Gaglione S, Smith BE, Kim EJ, Dong J, Ronsard L, Okonkwo V, Lingwood D, Dougan M, Dougan SK, Birnbaum ME. Nat Methods. 2022 Apr;19(4):449-460. doi: 10.1038/s41592-022-01436-z. Epub 2022 Apr 8. 10.1038/s41592-022-01436-z PubMed 35396484 |
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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 pMD2-VSVGmut 5822 bp ds-DNA circular SYN 05-JUL-2022
DEFINITION Expresses VSVGmut protein in mammalian cells for lentiviral
pseudotyping.
ACCESSION .
VERSION .
KEYWORDS pMD2-VSVGmut
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5822)
TITLE Antigen identification and high-throughput interaction mapping by
reprogramming viral entry
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5822)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jul 5, 2022 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5822
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(40..59)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
enhancer 161..540
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 541..744
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 695..715
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 741..765
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 878..1353
/label=-beta-globin intron
/note="internally truncated intron from human beta-globin"
CDS 1428..2963
/codon_start=1
/product="vesicular stomatitis virus G glycoprotein"
/label=VSV-G
/note="Indiana strain"
/translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
LNWHNDLIGTALQVKMPQSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQ
FINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFR
SNYFAYETGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLK
YFETRYIRVDIAAPILSRMVGMISGTTTEAELWDDWAPYEDVEIGPNGVLRTSSGYKFP
LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWF
SSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK"
polyA_signal 3164..3558
/label=-beta-globin poly(A)
/note="human beta-globin polyadenylation signal"
primer_bind complement(3897..3914)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4068..4656)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4148..4167)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4827..5687)
/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 5450..5469
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5688..5792)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ggatcccctg agggggcccc catgggctag aggatccggc ctcggcctct gcataaataa
61 aaaaaattag tcagccatga gcttggccca ttgcatacgt tgtatccata tcataatatg
121 tacatttata ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt
181 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
241 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
301 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
361 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
421 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
481 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
541 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
601 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
661 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
721 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
781 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccc ctcgaagctt
841 acatgtggta ccgagctcgg atcctgagaa cttcagggtg agtctatggg acccttgatg
901 ttttctttcc ccttcttttc tatggttaag ttcatgtcat aggaagggga gaagtaacag
961 ggtacacata ttgaccaaat cagggtaatt ttgcatttgt aattttaaaa aatgctttct
1021 tcttttaata tacttttttg tttatcttat ttctaatact ttccctaatc tctttctttc
1081 agggcaataa tgatacaatg tatcatgcct ctttgcacca ttctaaagaa taacagtgat
1141 aatttctggg ttaaggcaat agcaatattt ctgcatataa atatttctgc atataaattg
1201 taactgatgt aagaggtttc atattgctaa tagcagctac aatccagcta ccattctgct
1261 tttattttat ggttgggata aggctggatt attctgagtc caagctaggc ccttttgcta
1321 atcatgttca tacctcttat cttcctccca cagctcctgg gcaacgtgct ggtctgtgtg
1381 ctggcccatc actttggcaa agcacgtgag atctgaattc tgacactatg aagtgccttt
1441 tgtacttagc ctttttattc attggggtga attgcaagtt caccatagtt tttccacaca
1501 accaaaaagg aaactggaaa aatgttcctt ctaattacca ttattgcccg tcaagctcag
1561 atttaaattg gcataatgac ttaataggca cagccttaca agtcaaaatg ccccagagtc
1621 acaaggctat tcaagcagac ggttggatgt gtcatgcttc caaatgggtc actacttgtg
1681 atttccgctg gtatggaccg aagtatataa cacattccat ccgatccttc actccatctg
1741 tagaacaatg caaggaaagc attgaacaaa cgaaacaagg aacttggctg aatccaggct
1801 tccctcctca aagttgtgga tatgcaactg tgacggatgc cgaagcagtg attgtccagg
1861 tgactcctca ccatgtgctg gttgatgaat acacaggaga atgggttgat tcacagttca
1921 tcaacggaaa atgcagcaat tacatatgcc ccactgtcca taactctaca acctggcatt
1981 ctgactataa ggtcaaaggg ctatgtgatt ctaacctcat ttccatggac atcaccttct
2041 tctcagagga cggagagcta tcatccctgg gaaaggaggg cacagggttc agaagtaact
2101 actttgctta tgaaactgga ggcaaggcct gcaaaatgca atactgcaag cattggggag
2161 tcagactccc atcaggtgtc tggttcgaga tggctgataa ggatctcttt gctgcagcca
2221 gattccctga atgcccagaa gggtcaagta tctctgctcc atctcagacc tcagtggatg
2281 taagtctaat tcaggacgtt gagaggatct tggattattc cctctgccaa gaaacctgga
2341 gcaaaatcag agcgggtctt ccaatctctc cagtggatct cagctatctt gctcctaaaa
2401 acccaggaac cggtcctgct ttcaccataa tcaatggtac cctaaaatac tttgagacca
2461 gatacatcag agtcgatatt gctgctccaa tcctctcaag aatggtcgga atgatcagtg
2521 gaactaccac agaagccgaa ctgtgggatg actgggcacc atatgaagac gtggaaattg
2581 gacccaatgg agttctgagg accagttcag gatataagtt tcctttatac atgattggac
2641 atggtatgtt ggactccgat cttcatctta gctcaaaggc tcaggtgttc gaacatcctc
2701 acattcaaga cgctgcttcg caacttcctg atgatgagag tttatttttt ggtgatactg
2761 ggctatccaa aaatccaatc gagcttgtag aaggttggtt cagtagttgg aaaagctcta
2821 ttgcctcttt tttctttatc atagggttaa tcattggact attcttggtt ctccgagttg
2881 gtatccatct ttgcattaaa ttaaagcaca ccaagaaaag acagatttat acagacatag
2941 agatgaaccg acttggaaag taactcaaat cctgcacaac agattcttca tgtttggacc
3001 aaatcaactt gtgataccat gctcaaagag gcctcaatta tatttgagtt tttaattttt
3061 atgnaaaaaa aaaaaaaaaa cggaattcac cccaccagtg caggctgcct atcagaaagt
3121 ggtggctggt gtggctaatg ccctggccca caagtatcac taagctcgct ttcttgctgt
3181 ccaatttcta ttaaaggttc ctttgttccc taagtccaac tactaaactg ggggatatta
3241 tgaagggcct tgagcatctg gattctgcct aataaaaaac atttattttc attgcaatga
3301 tgtatttaaa ttatttctga atattttact aaaaagggaa tgtgggaggt cagtgcattt
3361 aaaacataaa gaaatgaaga gctagttcaa accttgggaa aatacactat atcttaaact
3421 ccatgaaaga aggtgaggct gcaaacagct aatgcacatt ggcaacagcc cctgatgcct
3481 atgccttatt catccctcag aaaaggattc aagtagaggc ttgatttgga ggttaaagtt
3541 ttgctatgct gtattttaca ttacttattg ttttagctgt cctcatgaat gtcttttcac
3601 tacccatttg cttatcctgc atctctcagc cttgactcca ctcagttctc ttgcttagag
3661 ataccacctt tcccctgaag tgttccttcc atgttttacg gcgagatggt ttctcctcgc
3721 ctggccactc agccttagtt gtctctgttg tcttatagag gtctacttga agaaggaaaa
3781 acagggggca tggtttgact gtcctgtgag cccttcttcc ctgcctcccc cactcacagt
3841 gacccggaat ccctcgacat ggcagtctag cactagtgcg gccgcagatc tgcttcctcg
3901 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
3961 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
4021 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
4081 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
4141 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
4201 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
4261 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
4321 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
4381 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
4441 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
4501 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
4561 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
4621 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
4681 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
4741 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
4801 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
4861 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
4921 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
4981 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
5041 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
5101 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
5161 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
5221 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
5281 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
5341 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
5401 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
5461 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
5521 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
5581 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
5641 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
5701 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
5761 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
5821 gt
//
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