| 质粒名称: | pJH373 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 86840 |
| 存储实验室: | Joseph Bondy-Denomy |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1(+) |
| 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: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | acrIIA2,LMOG_03147 |
| 物种来源: | Listeria monocytogenes |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 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 | CMV-F |
| 3' 测序引物 | BGH-Rev |
|
文献引用方法 |
|
| 材料和方法部分: | pJH373 was a gift from Joseph Bondy-Denomy (Addgene plasmid # 86840 ; http://n2t.net/addgene:86840 ; RRID:Addgene_86840) |
| 参考文献部分: | Inhibition of CRISPR-Cas9 with Bacteriophage Proteins. Rauch BJ, Silvis MR, Hultquist JF, Waters CS, McGregor MJ, Krogan NJ, Bondy-Denomy J. Cell. 2016 Dec 22. pii: S0092-8674(16)31683-X. doi: 10.1016/j.cell.2016.12.009. 10.1016/j.cell.2016.12.009 PubMed 28041849 |
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 pJH373 5792 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION mamalian expression of AcrIIA2.
ACCESSION .
VERSION .
KEYWORDS pJH373
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5792)
AUTHORS Rauch BJ, Silvis MR, Hultquist JF, Waters CS, McGregor MJ, Krogan
NJ, Bondy-Denomy J
TITLE Inhibition of CRISPR-Cas9 with Bacteriophage Proteins.
JOURNAL Cell. 2016 Dec 22. pii: S0092-8674(16)31683-X. doi:
10.1016/j.cell.2016.12.009.
PUBMED 28041849
REFERENCE 2 (bases 1 to 5792)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5792
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 42..421
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 422..625
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 576..596
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 670..689
/label=T7
/note="T7 promoter, forward primer"
promoter 670..688
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1190..1207)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1196..1420
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1466..1894
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1553..1572)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1763..1784
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(1903..1923)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 1908..2237
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2088..2223
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2150..2169
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2304..3098
/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(2358..2377)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 2968..2987
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3272..3393
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3309..3328)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3363..3382
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3442..3458)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3442..3458)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3455..3477)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3466..3482
/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(3490..3520)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3535..3556
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3673..3690)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3844..4432)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3924..3943)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4603..5463)
/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 5226..5245
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5464..5568)
/gene="bla"
/label=AmpR promoter
primer_bind complement(5643..5662)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
61 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
121 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
181 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
241 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
301 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
361 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
421 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
481 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
541 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
601 gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg cttactggct
661 tatcgaaatt aatacgactc actataggga gacccaagct ggctagcgtt taaacttaag
721 cttggtaccg agctcggatc cgccaccatg acgctgaccc gcgctcagaa aaaatacgcc
781 gaggcgatgc atgagtttat caatatggtt gatgactttg aagaatcaac gcctgacttt
841 gcaaaagagg ttctgcacga ctccgactat gtggtcatta caaaaaacga gaaatatgcc
901 gtggcactct gtagtctctc cacagatgaa tgtgagtacg atactaactt gtatttggat
961 gaaaagctcg tcgattacag cacagttgat gtcaacggag tgacatatta catcaatata
1021 gtggaaacaa atgacataga tgatcttgaa attgcgaccg acgaggacga gatgaagtct
1081 ggaaaccaag agattattct taagtccgaa ctgaagtaag aattctgcag atatccagca
1141 cagtggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
1201 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
1261 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
1321 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
1381 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
1441 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
1501 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
1561 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
1621 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
1681 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
1741 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
1801 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
1861 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
1921 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
1981 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
2041 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
2101 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
2161 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
2221 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
2281 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
2341 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
2401 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
2461 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
2521 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
2581 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
2641 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
2701 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
2761 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
2821 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
2881 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
2941 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
3001 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
3061 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
3121 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
3181 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
3241 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
3301 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
3361 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
3421 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
3481 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
3541 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
3601 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
3661 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
3721 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
3781 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
3841 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
3901 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
3961 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
4021 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
4081 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
4141 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
4201 tggtaacagg attagcagag cgaggtatgt aggcagtgct acagagttct tgaagtggtg
4261 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
4321 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
4381 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
4441 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
4501 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
4561 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
4621 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
4681 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
4741 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
4801 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
4861 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
4921 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
4981 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
5041 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
5101 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
5161 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
5221 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
5281 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
5341 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
5401 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
5461 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
5521 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
5581 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
5641 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
5701 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
5761 aattgcatga agaatctgct tagggttagg cg
//
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