| 质粒名称: | Mm-PylRS-AF/Pyl-tRNACUA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 122650 |
| 存储实验室: | Howard Hang |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 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: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | PylRS-AF, Pyl-tRNACUA |
| 物种来源: | Methanosarcina mazei |
| 插入大小: | -- |
| 基因突变: | 酪氨酸306变为丙氨酸，酪氨酸384变为苯丙氨酸 |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | FLAG |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | CGCAAATGGGCGGTAGGCGTG |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | Mm-PylRS-AF/Pyl-tRNACUA was a gift from Howard Hang (Addgene plasmid # 122650 ; http://n2t.net/addgene:122650 ; RRID:Addgene_122650) |
| 参考文献部分: | IFITM3 directly engages and shuttles incoming virus particles to lysosomes. Spence JS, He R, Hoffmann HH, Das T, Thinon E, Rice CM, Peng T, Chandran K, Hang HC. Nat Chem Biol. 2019 Jan 14. pii: 10.1038/s41589-018-0213-2. doi: 10.1038/s41589-018-0213-2. 10.1038/s41589-018-0213-2 [pii] PubMed 30643282 |
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 Mm-PylRS-AF/Pyl-tRNACUA 7401 bp ds-DNA circular SYN 05-MAR-2019
DEFINITION tRNA synthetase/tRNA pair for the incorporation of unnatural amino
acid into proteins in mammalian cells in response to the amber codon
TAG..
ACCESSION .
VERSION .
KEYWORDS Mm-PylRS-AF/Pyl-tRNACUA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7401)
AUTHORS Spence JS, He R, Hoffmann HH, Das T, Thinon E, Rice CM, Peng T,
Chandran K, Hang HC
TITLE IFITM3 directly engages and shuttles incoming virus particles to
lysosomes.
JOURNAL Nat Chem Biol. 2019 Jan 14. pii: 10.1038/s41589-018-0213-2. doi:
10.1038/s41589-018-0213-2.
PUBMED 30643282
REFERENCE 2 (bases 1 to 7401)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 5, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7401
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 46..425
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 426..629
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 580..600
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 626..650
/label=LNCX
/note="Human CMV promoter, forward primer"
promoter 860..878
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 860..877
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind 891..910
/label=pMT2-F
/note="Synthetic intron, forward primer"
regulatory 962..971
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 971..994
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
primer_bind complement(2385..2402)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2391..2615
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2661..3089
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2748..2767)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2958..2979
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3098..3118)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3103..3432
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3283..3418
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3345..3364
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3481..4506
/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 4636..4757
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4673..4692)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4727..4746
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4806..4822)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4806..4822)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4819..4841)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4830..4846
/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(4854..4884)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4899..4920
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5037..5054)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5208..5796)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5288..5307)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5967..6827)
/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 6590..6609
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6828..6932)
/gene="bla"
/label=AmpR promoter
promoter complement(7030..7270)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind complement(7080..7099)
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(7250..7270)
/label=hU6-F
/note="Human U6 promoter, forward primer"
ORIGIN
1 atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgttggcat tgattattga
61 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
121 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
181 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
241 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
301 caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
361 acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
421 ccatggtgat gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg
481 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
541 gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg
601 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
661 gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccgcggcc
721 gggaacggtg cattggaacg cggattcccc gtgccaagag tgacgtaagt accgcctata
781 gagtctatag gcccaccccc ttggcttcgt tagaacgcgg ctacaattaa tacataacct
841 tatgtatcat acacatacga tttaggtgac actatagaat aacatccact ttgcctttct
901 ctccacaggt gtccactccc aggtccaact gcacgggcta gcgtttaaac ttaagaagct
961 tgccaccatg gactacaagg acgacgacga caagatggac aagaagcccc tgaacaccct
1021 gatcagcgcc acaggactgt ggatgtccag aaccggcacc atccacaaga tcaagcacca
1081 cgaggtgtcc cggtccaaaa tctacatcga gatggcctgc ggcgatcacc tggtcgtcaa
1141 caacagcaga agcagccgga cagccagagc cctgcggcac cacaagtaca gaaagacctg
1201 caagcggtgc agagtgtccg acgaggacct gaacaagttc ctgaccaagg ccaacgagga
1261 ccagaccagc gtgaaagtga aggtggtgtc cgcccccacc cggaccaaga aagccatgcc
1321 caagagcgtg gccagagccc ccaagcccct ggaaaacacc gaagccgctc aggcccagcc
1381 cagcggcagc aagttcagcc ccgccatccc cgtgtctacc caggaaagcg tcagcgtccc
1441 cgccagcgtg tccaccagca tctctagcat ctcaaccggc gccacagctt ctgccctggt
1501 caagggcaac accaacccca tcaccagcat gtctgcccct gtgcaggcct ctgccccagc
1561 cctgaccaag tcccagaccg accggctgga agtgctcctg aaccccaagg acgagatcag
1621 cctgaacagc ggcaagccct tccgggagct ggaaagcgag ctgctgagcc ggcggaagaa
1681 ggacctccag caaatctacg ccgaggaacg ggagaactac ctgggcaagc tggaaagaga
1741 gatcacccgg ttcttcgtgg accggggctt cctggaaatc aagagcccca tcctgatccc
1801 cctggagtac atcgagcgga tgggcatcga caacgacacc gagctgagca agcagatttt
1861 ccgggtggac aagaacttct gcctgcggcc catgctggcc cccaacctgg ccaactacct
1921 gcggaaactg gatcgcgctc tgcccgaccc catcaagatt ttcgagatcg gcccctgcta
1981 ccggaaagag agcgacggca aagagcacct ggaagagttt acaatgctga acttttgcca
2041 gatgggcagc ggctgcacca gagagaacct ggaatccatc atcaccgact ttctgaacca
2101 cctggggatc gacttcaaga tcgtgggcga cagctgcatg gtgttcggcg acaccctgga
2161 cgtgatgcac ggcgacctgg aactgtctag cgccgtcgtg ggacccatcc ctctggaccg
2221 ggagtggggc atcgataagc cctggatcgg agccggcttc ggcctggaac ggctgctgaa
2281 agtcaagcac gactttaaga acatcaagcg ggctgccaga agcgagagct actacaacgg
2341 catcagcacc aacctgtgat gataaggatc cacccgctga tcagcctcga ctgtgccttc
2401 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
2461 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
2521 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
2581 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
2641 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
2701 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
2761 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
2821 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
2881 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
2941 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
3001 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
3061 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
3121 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
3181 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
3241 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
3301 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
3361 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
3421 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
3481 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
3541 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
3601 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
3661 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
3721 ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
3781 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat
3841 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
3901 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
3961 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
4021 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
4081 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
4141 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
4201 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg
4261 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
4321 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
4381 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc
4441 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
4501 gaatagcacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
4561 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
4621 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
4681 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
4741 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
4801 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
4861 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
4921 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
4981 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
5041 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
5101 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
5161 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
5221 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
5281 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
5341 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
5401 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
5461 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
5521 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
5581 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
5641 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
5701 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
5761 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
5821 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
5881 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
5941 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
6001 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
6061 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
6121 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
6181 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
6241 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
6301 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
6361 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
6421 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
6481 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
6541 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
6601 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
6661 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
6721 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
6781 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
6841 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
6901 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaaat ggcggaaacc
6961 ccgggaatct aacccggctg aacggattta gagtccattc gatctacatg atcaggtttc
7021 cggtgtttcg tcctttccac aagatatata aagccaagaa atcgaaatac tttcaagtta
7081 cggtaagcat atgatagtcc attttaaaac ataattttaa aactgcaaac tacccaagaa
7141 attattactt tctacgtcac gtattttgta ctaatatctt tgtgtttaca gtcaaattaa
7201 ttctaattat ctctctaaca gccttgtatc gtatatgcaa atatgaagga atcatgggaa
7261 ataggccctc ttcctgcccg accttcgcga tgtacgggcc agatatacgc caatattggc
7321 cattagccat attattcatt ggttatatag cataaatcaa tattggctat tggccattgc
7381 atacgttgta tctatatcat a
//
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