质粒名称: | RAB11A-GFP HDRT Source (pTR 143) |
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出品公司: | 爱迪基因 |
目录编号: | 112012 |
存储实验室: | Alexander Marson |
载体骨架基本信息 |
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载体名称: | pUC19 |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | 2686 bp |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | CRISPR,合成生物学 |
筛选标记: | -- |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | RAB11A-GFP HDRT |
物种来源: | H. sapiens (human) |
插入大小: | 1350 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | -- |
克隆方法: | Gibson克隆 |
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 |
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材料和方法部分: | RAB11A-GFP HDRT Source (pTR 143) was a gift from Alexander Marson (Addgene plasmid # 112012 ; http://n2t.net/addgene:112012 ; RRID:Addgene_112012) |
参考文献部分: | Reprogramming human T cell function and specificity with non-viral genome targeting. Roth TL, Puig-Saus C, Yu R, Shifrut E, Carnevale J, Li PJ, Hiatt J, Saco J, Krystofinski P, Li H, Tobin V, Nguyen DN, Lee MR, Putnam AL, Ferris AL, Chen JW, Schickel JN, Pellerin L, Carmody D, Alkorta-Aranburu G, Del Gaudio D, Matsumoto H, Morell M, Mao Y, Cho M, Quadros RM, Gurumurthy CB, Smith B, Haugwitz M, Hughes SH, Weissman JS, Schumann K, Esensten JH, May AP, Ashworth A, Kupfer GM, Greeley SAW, Bacchetta R, Meffre E, Roncarolo MG, Romberg N, Herold KC, Ribas A, Leonetti MD, Marson A. Nature. 2018 Jul 11. pii: 10.1038/s41586-018-0326-5. doi: 10.1038/s41586-018-0326-5. 10.1038/s41586-018-0326-5 PubMed 29995861 |
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LOCUS RAB11A-GFP HDRT Source (pTR 143) 5814 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION DNA sequence source for amplifying an HDR template to tag endogenous human RAB11A gene with GFP. ACCESSION . VERSION . KEYWORDS RAB11A-GFP HDRT Source (pTR 143) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5814) AUTHORS Roth TL, Puig-Saus C, Yu R, Shifrut E, Carnevale J, Li PJ, Hiatt J, Saco J, Krystofinski P, Li H, Tobin V, Nguyen DN, Lee MR, Putnam AL, Ferris AL, Chen JW, Schickel JN, Pellerin L, Carmody D, Alkorta-Aranburu G, Del Gaudio D, Matsumoto H, Morell M, Mao Y, Cho M, Quadros RM, Gurumurthy CB, Smith B, Haugwitz M, Hughes SH, Weissman JS, Schumann K, Esensten JH, May AP, Ashworth A, Kupfer GM, Greeley SAW, Bacchetta R, Meffre E, Roncarolo MG, Romberg N, Herold KC, Ribas A, Leonetti MD, Marson A TITLE Reprogramming human T cell function and specificity with non-viral genome targeting. JOURNAL Nature. 2018 Jul 11. pii: 10.1038/s41586-018-0326-5. doi: 10.1038/s41586-018-0326-5. PUBMED 29995861 REFERENCE 2 (bases 1 to 5814) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5814 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 235..254 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(473..577) /gene="bla" /label=AmpR promoter primer_bind 645..663 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(701..723) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 823..842 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1036..1058 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 1050..1067 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1051..1067 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 2309..2767 /codon_start=1 /product="improved N-terminal fragment of mVenus for use in bimolecular fluorescence complementation (BiFC) (Kodama and Hu, 2010)" /label=VN155(I152L) /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITA" primer_bind complement(2348..2369) /label=EGFP-N /note="EGFP, reverse primer" primer_bind 2956..2977 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(4265..4281) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4265..4281) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4278..4300) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4289..4305 /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(4313..4343) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4358..4379 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4496..4513) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4667..5255) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4747..4766) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5426..472) /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" ORIGIN 1 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 61 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 121 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 181 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 241 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 301 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 361 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 421 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 481 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 541 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 601 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga 661 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc 721 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg 781 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg 841 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc 901 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg 961 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg 1021 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttcgagctcg 1081 gtacccgggg agttataggc atacttgccc acaagtctaa atcgagagtt tgcatatagg 1141 aggtaggagg tgtgacaata attggataaa ggcttttctg gaatgacaaa attctcttca 1201 aatacaggga ggccctgcga catgatatct ggcaaatcac ttgagtctta gtatccaaac 1261 ttccgctaaa atggaaatta ccatcctttg cttggcatgt taaatgaaat acttatgtaa 1321 acggacttag attagaggcc tacaaatagg actcaagttc ttgatgaatc ttgatgagga 1381 gggtgggcga agtactctaa atgaaaatga gatagattgc gagatttccg gagaggagtg 1441 agtgacaagc gcctacccgt gcatgaagct ggaggtagaa aattccagca aaaggaattt 1501 tcaagtttca ctgaagcttt tgaatcaagg cttgaaagcc cagggttaaa aaaaaggcag 1561 tgaagaagct cattaagaca accccccgcc aggtagcaga tagcacctga aatgcagtgg 1621 gaaatttgtg aggtcttcat aggcaaggat taaattttac atctccatct taagatgtgt 1681 catagttgtt caattactta gtgaccaaaa aagtgaatta aagatgttgt caagattaaa 1741 aaaacaaaaa ccaatgcagg ttccaccgag atttgaactc ggatcgctgg attcagagtc 1801 cagagtgcta accattacac catggaaccg ccacgcatgt gtagctgcct tcggctgtct 1861 aatcctcaga gaaccccgcc cccatccaca aacccaccac tcacaggcgg tcccgcctgg 1921 ttccagcgag ccgcttccgg cacggtagct cgagaaatga gcaagcggcc actaagacta 1981 tggtagctag gagttccagg actcagtttc ccctttgagc ctcctttagc gactaaagct 2041 tgaagcccca cgcatctcga ctctcgcgca caccgccctt gttgggctca ggggcggggc 2101 gccgcccccg gaagtacttc cccttaaagg ctggggcctg ccggaaatgg cgcagcggca 2161 gggaggggct cttcacccag tccggcagtt gaagctcggc gctcgggtta cccctgcagc 2221 gacgccccct ggtcccacag ataccactgc tgctcccgcc ctttcgctcc tcggccgcgc 2281 aatgggcgga tcgggtggga ctagtggcag caagggcgag gagctgttca ccggggtggt 2341 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgcgcggcga 2401 gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa 2461 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag 2521 ccgctacccc gaccacatga agcgccacga cttcttcaag tccgccatgc ccgaaggcta 2581 cgtccaggag cgcaccatca gcttcaagga cgacggcacc tacaagaccc gcgccgaggt 2641 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 2701 ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca acgtctatat 2761 caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacgtgga 2821 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 2881 cgtgctgctg cccgacaacc actacctgag cacccagtcc gtgctgagca aagaccccaa 2941 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactggaac 3001 cggtgctgga agtggtacac gcgacgacga gtacgactac ctctttaaag gtgaggccat 3061 gggctctcgc actctacaca gtcctcgttc ggggacccgg gccactcccg gtggaccctc 3121 gtgccggcca cccctgcact gatataggcc tccctcagcc cttccttttt gtgcggttcc 3181 gtctcctacc cagctcagcc tcttctcccc cgctcagaca ggggtcccca tcacatgccg 3241 ctctctgagc gacctctcca taggccttcg ctggcctcag agcccctccc tgcgtgtcct 3301 tcccctggcg gactgccttc tcccacatcg tcgaattcct ttccccgggt tctacggccc 3361 cgccgctcct cccaccatct ctcttttcgg gtgtagcgcc ccctccccct cggcgtacac 3421 ccttcccagc tcgcgtcctc tcccgaagcc cctctgacgg gttcttcgct tccctcttgg 3481 ccttgccttc ggtgcagact cccattacag gtctttttct tatctgccct ctttcccccc 3541 tcgagcgagc gtctcttctg tatctcccct ccacttggac ggccgcccct ctaagcttct 3601 cccatcccgg gtccaggacc ccgtaaaatc tgagcgggcc aagggcctcc gcccccaccc 3661 tccgccccgc ggttctcgcg gtccttaacc ttctctgacc gctccctctc ccactcccca 3721 tttggcaact ctctctacct tccctcccca aatcgtgttt agagctccct tgtctccgtt 3781 gcgacctggg tggtgggagg gagggtaaga gggatcgccc atcacccact cgcgcacttg 3841 tcgcagtgac ccggcaggct gacattactt cccacctagc cttttctttt cttgccctcc 3901 ctactcagtg gcacccaacc cgggataact ctttactcgg ttcgcgagat gaggttgcgg 3961 atgggtggtg gctgttatcc ttgccagacc tgaaggcctg gttcctgagc aggaaaggtt 4021 gtgcgctgca caaatactag cttgctttat ctctatgaac tggttgcttc ctgctgcaac 4081 gctgggagga tccaacagga agggaagagc cctagttagc cacatacctt tcccaaaagg 4141 ttgggggtgg gaggaagttg tgtgctcttt tggttacatt aaatatttat tataagatac 4201 ggaaacatgt agtgcccttt cctctagagt cgacctgcag gcatgcaagc ttggcgtaat 4261 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 4321 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 4381 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 4441 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 4501 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 4561 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 4621 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 4681 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 4741 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 4801 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 4861 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 4921 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 4981 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 5041 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 5101 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 5161 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 5221 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 5281 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 5341 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 5401 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 5461 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 5521 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 5581 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 5641 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 5701 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 5761 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgag //
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