| 质粒名称: | pCSW21 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 86836 |
| 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: | 4643bp |
| 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: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | acrIIA4 / LMOG_02993 |
| 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: |
pCSW21 was a gift from Joseph Bondy-Denomy (Addgene plasmid # 86836 ; http://n2t.net/addgene:86836 ; RRID:Addgene_86836)
|
| 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 pCSW21 4643 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION bacterial expression of AcrIIA4.
ACCESSION .
VERSION .
KEYWORDS pCSW21
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4643)
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 4643)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4643
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 8..292
/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 215..234
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
terminator 678..764
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 856..883
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 902..993
/gene="bla"
/label=AmpR promoter
CDS 994..1854
/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(1212..1231)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1896..2351
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1983..2002)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2193..2214
/label=F1ori-F
/note="F1 origin, forward primer"
rep_origin 2462..3050
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2951..2970
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3204..3221
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 3387..3409
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 3547..3566
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
CDS complement(3746..4624)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
ORIGIN
1 ttcagagaag aaaccaattg tccatattgc atcagacatt gccgtcactg cgtcttttac
61 tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct gtaacaaagc
121 gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc
181 cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt atccataaga
241 ttagcggatc ctacctgacg ctttttatcg caactctcta ctgtttctcc atacccgttt
301 ttttgggcta gcaggaggaa ttcaccatga atattaatga cttaattaga gaaatcaaaa
361 acaaagatta cacagtgaaa ttgagtggta cggatagcaa tagtatcaca cagctaatta
421 ttcgcgttaa taatgatggc aacgagtatg taatttctga aagtgaaaat gaatcaatcg
481 ttgaaaaatt catctctgca ttcaaaaacg gttggaatca agaatacgag gatgaagaag
541 aattttataa tgacatgcaa acaatcacct taaaaagtga gttgaactaa aaatatgtca
601 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt
661 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
721 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
781 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
841 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc
901 ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
961 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
1021 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
1081 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
1141 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
1201 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
1261 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
1321 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
1381 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
1441 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
1501 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
1561 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
1621 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
1681 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
1741 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
1801 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
1861 gaccaagttt actcatatat actttagatt gatttacgcg ccctgtagcg gcgcattaag
1921 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
1981 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
2041 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
2101 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
2161 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
2221 actcaaccct atctcgggct attcttttga tttataaggg attttgccga tttcggccta
2281 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
2341 gtttacaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
2401 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
2461 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
2521 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
2581 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
2641 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
2701 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
2761 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
2821 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
2881 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
2941 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
3001 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
3061 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
3121 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
3181 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
3241 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataggg tcatggctgc
3301 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
3361 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
3421 atcaccgaaa cgcgcgaggc agaaggagat ggcgcccaac agtcccccgg ccacggggcc
3481 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
3541 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
3601 ggccacgatg cgtccggcgt agaggatctg ctcatgtttg acagcttatc atcgatgcat
3661 aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac tccgtcaagc
3721 cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca ttcacttttt
3781 cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg
3841 agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg
3901 tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc
3961 taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct
4021 gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag
4081 cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc
4141 gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt
4201 gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg
4261 ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg
4321 cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt
4381 agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc
4441 gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca
4501 ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac
4561 ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag
4621 ccatactttt catactcccg cca
//
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