| 质粒名称: | pEJS443-pCSDest2-AcrIIC3Nme |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 85713 |
| 存储实验室: | Erik Sontheimer |
|
载体骨架基本信息 |
|
| 载体名称: | pCSDest2 |
| 空载体大小: | 4097 bp |
| 完整质粒大小: | 4448bp |
| 载体类型: | 哺乳细胞表达、CRISPR |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | Type II-C anti-CRISPR AcrIIC3Nme |
| 物种来源: | Neisseria meningitidis |
| 插入大小: | 351 bp |
| GenBank编号 | WP_042743676.1 |
| 启动子: | CMV |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | CGC AAA TGG GCG GTA GGC GTG |
| 3’测序引物 | CAG GAA ACA GCT ATG AC |
|
文献引用方法 |
|
| 材料和方法部分: |
pEJS443-pCSDest2-AcrIIC3Nme was a gift from Erik Sontheimer (Addgene plasmid # 85713 ; http://n2t.net/addgene:85713 ; RRID:Addgene_85713)
|
| 参考文献部分: | Naturally Occurring Off-Switches for CRISPR-Cas9. Pawluk A, Amrani N, Zhang Y, Garcia B, Hidalgo-Reyes Y, Lee J, Edraki A, Shah M, Sontheimer EJ, Maxwell KL, Davidson AR. Cell. 2016 Dec 15;167(7):1829-1838.e9. doi: 10.1016/j.cell.2016.11.017. Epub 2016 Dec 8. 10.1016/j.cell.2016.11.017 PubMed 27984730 |
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 pEJS443-pCSDest2-AcrIIC3Nme 4445 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Mammalian expression of Type II-C anti-CRISPR protein AcrIIC3Nme
from Neisseria meningitidis.
ACCESSION .
VERSION .
KEYWORDS pEJS443-pCSDest2-AcrIIC3Nme
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4445)
AUTHORS Pawluk A, Amrani N, Zhang Y, Garcia B, Hidalgo-Reyes Y, Lee J,
Edraki A, Shah M, Sontheimer EJ, Maxwell KL, Davidson AR
TITLE Naturally Occurring Off-Switches for CRISPR-Cas9.
JOURNAL Cell. 2016 Dec 15;167(7):1829-1838.e9. doi:
10.1016/j.cell.2016.11.017. Epub 2016 Dec 8.
PUBMED 27984730
REFERENCE 2 (bases 1 to 4445)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4445
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 914..932
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 914..931
/label=SP6
/note="SP6 promoter, forward primer"
promoter complement(1354..1371)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 1376..1510
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1400..1419)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1454..1473
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(1623..1643)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(1623..1641)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1662..1678)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1662..1678)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1675..1697)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1686..1702
/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(1710..1740)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1755..1776
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1893..1910)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2064..2652)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2144..2163)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2823..3683)
/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 3446..3465
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3684..3788)
/gene="bla"
/label=AmpR promoter
rep_origin complement(3814..4269)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3951..3972)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4163..4182
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4347..879
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
ORIGIN
1 atatggtgga tctggacccc agccaattca atatggcgga cttggcacca tgccaattca
61 atatggcgga cctggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt
121 tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa
181 tggcggccat attggctata tgccaggatc aatatatagg caatatccaa tatggcccta
241 tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt
301 ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc
361 atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta
421 tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc
481 ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg
541 accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc
601 ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata
661 gtaacttggc aagtacatta ctattggaag tacgccaggg tacattggca gtactcccat
721 tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa
781 tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc
841 taatgggagg tctatataag caatgctcgt ttagggaacc gccattctgc ctggggacgt
901 cggagcaagc ttgatttagg tgacactata gaatacaagc tacttgttct ttttgcagga
961 tcccatcgat tcgaagccgc caccatgttc aaacgcgcta ttatcttcac ttctttcaac
1021 ggctttgaaa aagtttctcg aactgaaaaa cgccgccttg ccaaaatcat caatgctcga
1081 gtttccatca tcgacgaata cttgagagcc aaagacacca acgcatcgct tgacggtcag
1141 taccgcgctt tcttgttcaa cgacgaatcg cccgcaatga ccgaatttct ggcaaaactt
1201 aaagcctttg ccgaaagttg caccggaatc agcatcgacg catgggaaat tgaagaaagc
1261 gaatacgtcc gcctgccggt ggaacgcagg gatttcttag cggcagccaa cggcaaagag
1321 atttttaaaa tttaatgact cgagcctcta gaactatagt gagtcgtatt acgtagatcc
1381 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
1441 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
1501 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
1561 ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt
1621 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
1681 tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa
1741 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
1801 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
1861 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
1921 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
1981 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
2041 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
2101 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
2161 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
2221 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
2281 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
2341 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
2401 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
2461 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
2521 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
2581 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
2641 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
2701 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
2761 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
2821 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
2881 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
2941 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
3001 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
3061 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
3121 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
3181 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
3241 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
3301 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
3361 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
3421 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
3481 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
3541 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
3601 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
3661 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
3721 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
3781 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
3841 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
3901 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
3961 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
4021 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
4081 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
4141 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
4201 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
4261 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
4321 cctcttcgct attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca
4381 attcaatatg gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca
4441 attca
//
