| 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
MmXYRS-4xM15载体质粒基本信息
MmXYRS-4xM15质粒图谱载体图谱和MmXYRS-4xM15载体序列质粒序列多克隆位点信息
MmXYRS-4xM15质粒载体简介
MmXYRS-4xM15质粒序列载体序列
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
//
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MmXYRS-4xM15质粒载体应用举例
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