| 质粒名称: | pCMV-MbPylRS(DiZPK) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 91706 |
| 存储实验室: | P. Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 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: | pylS |
| 物种来源: | Methanosarcina barkeri |
| 插入大小: | 1260 bp |
| 基因突变: | T810G, C813T, C820G, T821C, T937A, A1046T, G1257A |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 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: | pCMV-MbPylRS(DiZPK) was a gift from P. Chen (Addgene plasmid # 91706 ; http://n2t.net/addgene:91706 ; RRID:Addgene_91706) |
| 参考文献部分: | Site-specific engineering of chemical functionalities on the surface of live hepatitis D virus. Lin S, Yan H, Li L, Yang M, Peng B, Chen S, Li W, Chen PR. Angew Chem Int Ed Engl. 2013 Dec 23;52(52):13970-4. doi: 10.1002/anie.201305787. Epub 2013 Nov 12. 10.1002/anie.201305787 PubMed 24222614 |
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
载体图谱.png)
载体简介
载体序列
LOCUS pCMV-MbPylRS(DiZPK) 7238 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION express Methanosarcina barkeri pyrrolysyl-tRNA synthetase/tRNA pair
in mammalian cells to incorporate the photocrosslinkers DiZPK,
DiZSeK, or DiZHSeC into amber codon site on a protein of interest.
ACCESSION .
VERSION .
KEYWORDS pCMV-MbPylRS(DiZPK)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7238)
AUTHORS Lin S, Yan H, Li L, Yang M, Peng B, Chen S, Li W, Chen PR
TITLE Site-specific engineering of chemical functionalities on the surface
of live hepatitis D virus.
JOURNAL Angew Chem Int Ed Engl. 2013 Dec 23;52(52):13970-4. doi:
10.1002/anie.201305787. Epub 2013 Nov 12.
PUBMED 24222614
REFERENCE 2 (bases 1 to 7238)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7238
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 63..442
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 443..646
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 597..617
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 643..667
/label=LNCX
/note="Human CMV promoter, forward primer"
promoter 877..895
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 877..894
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind 908..927
/label=pMT2-F
/note="Synthetic intron, forward primer"
regulatory 960..969
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind complement(2239..2256)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2245..2469
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2515..2943
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2602..2621)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2812..2833
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2952..2972)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2957..3286
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3137..3272
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3199..3218
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3335..4360
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
polyA_signal 4490..4611
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4527..4546)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4581..4600
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4660..4676)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4660..4676)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4673..4695)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4684..4700
/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(4708..4738)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4753..4774
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4891..4908)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5062..5650)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5142..5161)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5821..6681)
/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 6444..6463
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6682..6786)
/gene="bla"
/label=AmpR promoter
promoter complement(6884..7124)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind complement(6934..6953)
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(7104..7124)
/label=hU6-F
/note="Human U6 promoter, forward primer"
ORIGIN
1 gttgtatcta tatcataata tgtacattta tattggctca tgtccaatat gaccgccatg
61 ttggcattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
121 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
181 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
241 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
301 tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc
361 ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt
421 attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata
481 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
541 ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca
601 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg
661 tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg
721 atccagcctc cgcggccggg aacggtgcat tggaacgcgg attccccgtg ccaagagtga
781 cgtaagtacc gcctatagag tctataggcc cacccccttg gcttcgttag aacgcggcta
841 caattaatac ataaccttat gtatcataca catacgattt aggtgacact atagaataac
901 atccactttg cctttctctc cacaggtgtc cactcccagg tccaactgca cggaagcttg
961 ccaccatgga taaaaaacca ttagatgttt taatatctgc gaccgggctc tggatgtcca
1021 ggactggcac gctccacaaa atcaagcacc atgaggtctc aagaagtaaa atatacattg
1081 aaatggcgtg tggagaccat cttgttgtga ataattccag gagttgtaga acagccagag
1141 cattcagaca tcataagtac agaaaaacct gcaaacgatg tagggtttcg gacgaggata
1201 tcaataattt tctcacaaga tcaaccgaaa gcaaaaacag tgtgaaagtt agggtagttt
1261 ctgctccaaa ggtcaaaaaa gctatgccga aatcagtttc aagggctccg aagcctctgg
1321 aaaattctgt ttctgcaaag gcatcaacga acacatccag atctgtacct tcgcctgcaa
1381 aatcaactcc aaattcgtct gttcccgcat cggctcctgc tccttcactt acaagaagcc
1441 agcttgatag ggttgaggct ctcttaagtc cagaggataa aatttctcta aatatggcaa
1501 agcctttcag ggaacttgag cctgaacttg tgacaagaag aaaaaacgat tttcagcggc
1561 tctataccaa tgatagagaa gactacctcg gtaaactcga acgtgatatt acgaaatttt
1621 tcgtagaccg gggttttctg gagataaagt ctcctatcct tattccggcg gaatacgtgg
1681 agagaatggg tattaataat gatactgaac tttcaaaaca gatcttccgg gtggataaaa
1741 atctctgctt gaggccaatg cttgccccga ctctgtataa ctatgcgcga aaactcgata
1801 ggattttacc aggcccaata aaaattttcg aagtcggacc ttgttaccgg aaagagtctg
1861 acggcaaaga gcacctggaa gaatttacta tggtgaactt cagtcagatg ggttcgggat
1921 gtactcggga aaatcttgaa gctctcatca aagagtttct ggactatctg gaaatcgact
1981 tcgaaatcgt aggagattcc tgtatggtct ttggggatac tcttgatata atgcacgggg
2041 acctggagct ttcttcggca gtcgtcgggc cagtttctct tgatagagaa tggggtattg
2101 acaaaccatg gataggtgca ggttttggtc ttgaacgctt gctcaaggtt atgcacggct
2161 ttaaaaacat taagagggca tcaaggtccg aatcttacta taatgggatt tcaaccaatc
2221 tataaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
2281 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
2341 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
2401 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
2461 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
2521 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
2581 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
2641 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
2701 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
2761 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
2821 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
2881 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
2941 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
3001 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
3061 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
3121 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
3181 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
3241 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
3301 ttgtatatcc attttcggat ctgatcagca cgtgatgaaa aagcctgaac tcaccgcgac
3361 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
3421 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
3481 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
3541 ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta
3601 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
3661 cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca
3721 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
3781 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
3841 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
3901 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
3961 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
4021 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
4081 cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat
4141 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
4201 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
4261 cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
4321 aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtgctac gagatttcga
4381 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
4441 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
4501 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
4561 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
4621 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
4681 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
4741 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
4801 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
4861 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
4921 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
4981 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
5041 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
5101 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
5161 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
5221 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
5281 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
5341 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
5401 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
5461 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
5521 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
5581 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
5641 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
5701 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
5761 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
5821 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
5881 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
5941 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
6001 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
6061 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
6121 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
6181 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
6241 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
6301 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
6361 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
6421 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
6481 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
6541 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
6601 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
6661 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
6721 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
6781 tccgcgcaca tttccccgaa aaatggcgga aaccccggga atctaacccg gctgaacgga
6841 tttagagtcc attcgatcta catgatcagg tttccggtgt ttcgtccttt ccacaagata
6901 tataaagcca agaaatcgaa atactttcaa gttacggtaa gcatatgata gtccatttta
6961 aaacataatt ttaaaactgc aaactaccca agaaattatt actttctacg tcacgtattt
7021 tgtactaata tctttgtgtt tacagtcaaa ttaattctaa ttatctctct aacagccttg
7081 tatcgtatat gcaaatatga aggaatcatg ggaaataggc cctcttcctg cccgaccttc
7141 gcgatgtacg ggccagatat acgccaatat tggccattag ccatattatt cattggttat
7201 atagcataaa tcaatattgg ctattggcca ttgcatac
//
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