| 质粒名称: | pJH372 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 86839 |
| 存储实验室: | Joseph Bondy-Denomy |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1(+) |
| 完整质粒大小: | 5870bp |
| 载体类型: | 哺乳细胞表达 |
| 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: | acrIIA1 / LMOG_03146 |
| 物种来源: | Listeria monocytogenes |
| 启动子: | CMV |
| 克隆方法: | Gibson Cloning |
| 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: | pJH372 was a gift from Joseph Bondy-Denomy (Addgene plasmid # 86839 ; http://n2t.net/addgene:86839 ; RRID:Addgene_86839) |
| 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 pJH372 5870 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION mamalian expression of AcrIIA1.
ACCESSION .
VERSION .
KEYWORDS pJH372
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5870)
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 5870)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5870
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 17..396
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 397..600
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 551..571
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 645..664
/label=T7
/note="T7 promoter, forward primer"
promoter 645..663
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1243..1260)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1249..1473
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1519..1947
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1606..1625)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1816..1837
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(1956..1976)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 1961..2290
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2141..2276
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2203..2222
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2357..3151
/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(2411..2430)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3021..3040
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3325..3446
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3362..3381)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3416..3435
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3495..3511)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3495..3511)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3508..3530)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3519..3535
/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(3543..3573)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3588..3609
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3726..3743)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3897..4485)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3977..3996)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4656..5516)
/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 5279..5298
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5517..5621)
/gene="bla"
/label=AmpR promoter
primer_bind complement(5696..5715)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
61 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
121 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
181 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
241 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
301 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
361 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
421 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
481 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
541 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
601 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
661 agggagaccc aagctggcta gcgtttaaac ttaagcttgg taccgagctc ggatccgcca
721 ccatgaccat caagttgctc gacgaatttc tcaagaaaca tgatcttacc cggtaccaac
781 tgtctaaact gactggcatt tctcaaaata ccttgaaaga tcagaatgaa aagcccttga
841 ataagtacac tgtatccata ctcagaagtc tcagcttgat ttccggtctg agcgttagcg
901 atgtactttt tgaacttgaa gacatagaaa agaattctga tgacttggct ggattcaagc
961 acctcctcga taaatacaaa ctctccttcc cagcacaaga atttgaactc tactgcctta
1021 ttaaagaatt cgagtccgca aatatcgaag ttcttccatt cacgtttaat agattcgaga
1081 acgaggaaca tgtgaatatt aagaaggatg tctgcaaggc tcttgaaaat gccatcaccg
1141 tcctgaagga aaaaaaaaat gagctgcttt gagaattctg cagatatcca gcacagtggc
1201 ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
1261 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
1321 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
1381 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
1441 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
1501 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
1561 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
1621 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
1681 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
1741 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
1801 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
1861 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
1921 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
1981 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
2041 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
2101 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
2161 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
2221 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
2281 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
2341 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
2401 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
2461 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
2521 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
2581 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
2641 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
2701 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
2761 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
2821 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
2881 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
2941 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
3001 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
3061 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
3121 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
3181 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
3241 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
3301 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
3361 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
3421 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
3481 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
3541 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
3601 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
3661 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
3721 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
3781 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
3841 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
3901 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
3961 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
4021 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
4081 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
4141 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
4201 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
4261 aggattagca gagcgaggta tgtaggcagt gctacagagt tcttgaagtg gtggcctaac
4321 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
4381 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
4441 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
4501 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
4561 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
4621 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
4681 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
4741 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
4801 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
4861 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
4921 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
4981 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
5041 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
5101 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
5161 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
5221 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
5281 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
5341 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
5401 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
5461 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
5521 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
5581 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
5641 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
5701 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
5761 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
5821 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
//
