质粒名称: | MmXYRS-4xM15 |
---|---|
出品公司: | Addgene |
目录编号: | 155343 |
存储实验室: | Irene Coin |
载体骨架基本信息 |
|
载体名称: | pcDNA3.1 |
载体出品公司: | Invitrogen, major modifications |
空载体大小: | -- |
完整质粒大小: | 8053 bp |
载体修饰: | -- |
载体类型: | 哺乳动物表达 |
筛选标记: | 新霉素(G418筛选) |
载体抗性: | 氨苄青霉素,100μg/mL |
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: | Humanized MmXYRS/tRNAM15 |
物种来源: | Synthetic; M. mazei |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子: | CMV/U6 |
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: | MmXYRS-4xM15 was a gift from Irene Coin (Addgene plasmid # 155343 ; http://n2t.net/addgene:155343 ; RRID:Addgene_155343) |
参考文献部分: | Exploring GPCR-arrestin interfaces with genetically encoded crosslinkers. Bottke T, Ernicke S, Serfling R, Ihling C, Burda E, Gurevich VV, Sinz A, Coin I. EMBO Rep. 2020 Sep 14:e50437. doi: 10.15252/embr.202050437. 10.15252/embr.202050437 PubMed 32929862 |
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 MmXYRS-4xM15 8053 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Plasmid encoding the translational pair that enables efficient incorporation of multiple phenylalanine-based amino acids in response to the amber codon into proteins in mammmalian cells.. ACCESSION . VERSION . KEYWORDS MmXYRS-4xM15 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8053) AUTHORS Bottke T, Ernicke S, Serfling R, Ihling C, Burda E, Gurevich VV, Sinz A, Coin I TITLE Exploring GPCR-arrestin interfaces with genetically encoded crosslinkers. JOURNAL EMBO Rep. 2020 Sep 14:e50437. doi: 10.15252/embr.202050437. PUBMED 32929862 REFERENCE 2 (bases 1 to 8053) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..8053 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 629..648 /label=T7 /note="T7 promoter, forward primer" promoter 629..647 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 667..676 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(2064..2081) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2070..2294 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2340..2768 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2427..2446) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2637..2658 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2777..2797) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2782..3111 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2962..3097 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3024..3043 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3178..3972 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3232..3251) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3842..3861 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4146..4267 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4183..4202) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4237..4256 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4314..4330) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4314..4330) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4327..4349) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4338..4354 /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(4362..4392) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4407..4428 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4545..4562) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4716..5304) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4796..4815) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5475..6335) /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 6098..6117 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6336..6440) /gene="bla" /label=AmpR promoter promoter complement(6558..6798) /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" promoter complement(6897..7137) /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" promoter complement(7236..7476) /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" promoter complement(7575..7815) /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind complement(7863..7882) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag 601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg 661 gctagcgcca ccatggacaa gaagcccctg aacaccctga tcagcgccac cggcctgtgg 721 atgagcagaa ccggcaccat ccacaagatc aagcaccacg aggtgtcccg gtccaagatc 781 tacatcgaga tggcctgcgg cgaccacctg gtcgtgaaca acagcagaag cagccggacc 841 gccagagccc tgagacacca caagtacaga aagacctgca agcggtgccg ggtgtccgac 901 gaggacctga acaagttcct gaccaaggcc aacgaggacc agaccagcgt gaaagtgaag 961 gtggtgtccg cccccacccg gaccaagaaa gccatgccta agagcgtggc cagggccccc 1021 aagcccctgg aaaacacaga agccgctcag gcccagccca gcggcagcaa attttctcct 1081 gccatccccg tgtccaccca ggaatccgtg tctgtgcctg cctccgtgtc taccagcatc 1141 agcagcatct ctaccggcgc cacagccagc gctctcgtga agggcaacac caaccccatc 1201 accagcatga gcgcccctgt gcaggcttct gcccctgccc tgacaaagag ccagaccgac 1261 agactggaag tgctgctgaa ccccaaggac gagatcagcc tgaacagcgg caagcctttc 1321 agagagctgg aaagcgagct gctgagccgg cggaagaagg acctgcagca gatctacgcc 1381 gaggaaagag agaactacct gggcaagctg gaacgggaaa tcacccggtt cttcgtggac 1441 cggggcttcc tggaaatcaa gagccccatc ctgatccccc tggagtacat cgagcggatg 1501 ggcatcgaca acgacaccga gctgagcaag cagatcttca gggtggacaa aaacttctgc 1561 ctgcggccca tgctgacccc caacctgtac aactatctgc ggaagctgga tagagccctg 1621 cccgacccca tcaagatctt cgagatcggc ccctgctaca gaaaagagag cgacggcaaa 1681 gagcacctgg aagagttcac catgctggcc ttcgcccaga tgggcagcgg ctgcaccaga 1741 gagaacctgg aatctatcat caccgacttt ctgaaccacc tggggatcga cttcaagatc 1801 gtgggcgaca gctgcatggt gttcggcgac accctggacg tgatgcacgg cgacctggaa 1861 ctgtctagcg ccgtcgtggg acccatccct ctggacagag agtggggcat cgataagccc 1921 accatcggag ccggcttcgg cctggaaaga ctgctgaaag tgaaacacga tttcaagaac 1981 atcaagaggg ccgccagaag cgagagctac tacaacggca tcagcaccaa cctgtgagcg 2041 gccgctttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt 2101 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta 2161 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg 2221 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc 2281 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca 2341 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 2401 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 2461 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag 2521 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 2581 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 2641 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 2701 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 2761 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca 2821 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc 2881 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata 2941 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 3001 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag 3061 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg 3121 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg 3181 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc 3241 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg 3301 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag 3361 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc 3421 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat 3481 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg 3541 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc 3601 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag 3661 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc 3721 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc 3781 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata 3841 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc 3901 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac 3961 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc 4021 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt 4081 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc 4141 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 4201 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 4261 tatcttatca tgtctgtata ccggtcctct agctagagct tggcgtaatc atggtcatag 4321 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 4381 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 4441 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 4501 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 4561 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 4621 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 4681 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 4741 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 4801 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 4861 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 4921 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 4981 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 5041 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 5101 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 5161 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 5221 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 5281 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 5341 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 5401 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 5461 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 5521 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 5581 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 5641 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 5701 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 5761 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 5821 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 5881 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 5941 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 6001 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 6061 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 6121 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 6181 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 6241 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 6301 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 6361 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 6421 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacaaaaaa 6481 cggaaacccc gggaatcgaa cccggctgaa cggatttaga gtccgttcgg tctccctgac 6541 caggtttccg gtgtttcgtc ctttccacaa gatatataaa gccaagaaat cgaaatactt 6601 tcaagttacg gtaagcatat gatagtccat tttaaaacat aattttaaaa ctgcaaacta 6661 cccaagaaat tattactttc tacgtcacgt attttgtact aatatctttg tgtttacagt 6721 caaattaatt ctaattatct ctctaacagc cttgtatcgt atatgcaaat atgaaggaat 6781 catgggaaat aggccctctt cctgcccctc gacaaaaaac ggaaaccccg ggaatcgaac 6841 ccggctgaac ggatttagag tccgttcggt ctccctgacc aggtttccgg tgtttcgtcc 6901 tttccacaag atatataaag ccaagaaatc gaaatacttt caagttacgg taagcatatg 6961 atagtccatt ttaaaacata attttaaaac tgcaaactac ccaagaaatt attactttct 7021 acgtcacgta ttttgtacta atatctttgt gtttacagtc aaattaattc taattatctc 7081 tctaacagcc ttgtatcgta tatgcaaata tgaaggaatc atgggaaata ggccctcttc 7141 ctgcccctcg acaaaaaacg gaaaccccgg gaatcgaacc cggctgaacg gatttagagt 7201 ccgttcggtc tccctgacca ggtttccggt gtttcgtcct ttccacaaga tatataaagc 7261 caagaaatcg aaatactttc aagttacggt aagcatatga tagtccattt taaaacataa 7321 ttttaaaact gcaaactacc caagaaatta ttactttcta cgtcacgtat tttgtactaa 7381 tatctttgtg tttacagtca aattaattct aattatctct ctaacagcct tgtatcgtat 7441 atgcaaatat gaaggaatca tgggaaatag gccctcttcc tgcccctcga caaaaaacgg 7501 aaaccccggg aatcgaaccc ggctgaacgg atttagagtc cgttcggtct ccctgaccag 7561 gtttccggtg tttcgtcctt tccacaagat atataaagcc aagaaatcga aatactttca 7621 agttacggta agcatatgat agtccatttt aaaacataat tttaaaactg caaactaccc 7681 aagaaattat tactttctac gtcacgtatt ttgtactaat atctttgtgt ttacagtcaa 7741 attaattcta attatctctc taacagcctt gtatcgtata tgcaaatatg aaggaatcat 7801 gggaaatagg ccctcttcct gcccctcgag tctagacccg atcccctatg gtgcactctc 7861 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg 7921 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac 7981 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc 8041 agatatacgc gtt //
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇:pCN-PDC1-GFP
- 下一篇:WNK2 WT