| 质粒名称: | pRAU171 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 86835 |
| 存储实验室: | Joseph Bondy-Denomy |
|
载体骨架基本信息 |
|
| 载体名称: | pBAD24 |
| 完整质粒大小: | 5668 bp |
| 载体类型: | 细菌细胞表达 |
| 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: | acrIIA2 / LMOG_03147 |
| 物种来源: | Listeria monocytogenes |
| 启动子: | araBAD |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | pBAD-F |
| 3’测序引物 | pBAD-R |
|
文献引用方法 |
|
| 材料和方法部分: |
pRAU171 was a gift from Joseph Bondy-Denomy (Addgene plasmid # 86835 ; http://n2t.net/addgene:86835 ; RRID:Addgene_86835)
|
| 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 pRAU171 5668 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION bacterial expression of AcrIIA2.
ACCESSION .
VERSION .
KEYWORDS pRAU171
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5668)
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 5668)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5668
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 16..300
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
primer_bind 223..242
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
terminator 799..885
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 977..1004
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 1023..1114
/gene="bla"
/label=AmpR promoter
terminator 1711..1797
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 1889..1916
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 1935..2026
/gene="bla"
/label=AmpR promoter
CDS 2027..2887
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(2245..2264)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2929..3384
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3016..3035)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3226..3247
/label=F1ori-F
/note="F1 origin, forward primer"
rep_origin 3495..4083
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3984..4003
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4237..4254
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 4420..4442
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 4580..4599
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
CDS complement(4779..5657)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
ORIGIN
1 tcccgccatt cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg
61 tcttttactg gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt
121 aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca
181 gaaaagtcca cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat
241 ccataagatt agcggatcct acctgacgct ttttatcgca actctctact gtttctccat
301 acccgttttt ttgggctagc aggaggaatt caccatgaca ttaacaagag cacaaaaaaa
361 gtatgcagaa gcgatgcacg agtttataaa tatggtggat gactttgaag agtctacacc
421 ggattttgca aaggaagttc tacatgattc tgactatgta gttattacaa aaaatgaaaa
481 atatgcagta gctctttgct ctcttagcac tgatgaatgt gaatatgata ctaacttata
541 cttagatgaa aaattggttg attactcaac agttgacgta aacggtgtga catactacat
601 caacattgtt gaaactaacg atatcgatga tttagaaatc gctacggatg aagatgagat
661 gaaaagtggc aaccaagaaa ttattttaaa aagtgagttg aagtaaaaaa tatgactatt
721 agaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
781 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
841 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
901 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
961 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact
1021 cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
1081 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
1141 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
1201 gaaagtctcc atacccgttt ttttgggcta gcaggaggaa ttcaccatga cattaacaag
1261 agcacaaaaa aagtatgcag aagcgatgca cgagtttata aatatggtgg atgactttga
1321 agagtctaca ccggattttg caaaggaagt tctacatgat tctgactatg tagttattac
1381 aaaaaatgaa aaatatgcag tagctctttg ctctcttagc actgatgaat gtgaatatga
1441 tactaactta tacttagatg aaaaattggt tgattactca acagttgacg taaacggtgt
1501 gacatactac atcaacattg ttgaaactaa cgatatcgat gatttagaaa tcgctacgga
1561 tgaagatgag atgaaaagtg gcaaccaaga aattatttta aaaagtgagt tgaagtaaaa
1621 aatatgacta ttagaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc
1681 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
1741 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
1801 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
1861 ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg
1921 tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
1981 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
2041 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
2101 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
2161 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
2221 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
2281 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
2341 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
2401 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
2461 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
2521 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
2581 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
2641 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
2701 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
2761 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
2821 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
2881 ttggtaactg tcagaccaag tttactcata tatactttag attgatttac gcgccctgta
2941 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
3001 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
3061 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
3121 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
3181 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
3241 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc
3301 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
3361 acaaaatatt aacgtttaca atttaaaagg atctaggtga agatcctttt tgataatctc
3421 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
3481 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
3541 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
3601 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
3661 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
3721 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
3781 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
3841 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
3901 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
3961 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
4021 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
4081 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
4141 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
4201 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
4261 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
4321 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
4381 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
4441 ggttttcacc gtcatcaccg aaacgcgcga ggcagaagga gatggcgccc aacagtcccc
4501 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
4561 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
4621 cgccggtgat gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt
4681 atcatcgatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc
4741 tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca
4801 tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt
4861 ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg
4921 ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc
4981 cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac
5041 aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg
5101 atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc
5161 gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag
5221 cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg
5281 tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat
5341 tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc
5401 ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg
5461 caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga
5521 atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca
5581 atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca
5641 ttttgcgctt cagccatact tttcatac
//
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