| 质粒名称: | pEJS433-pCSDest2-AcrIIC1Nme |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 85679 |
| 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: | 4097bp |
| 完整质粒大小: | 4352 bp |
| 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 AcrIIC1Nme |
| 物种来源: | Neisseria meningitidis |
| 插入大小: | 255bp |
| GenBank编号 | WP_049360089.1 |
| 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: |
pEJS433-pCSDest2-AcrIIC1Nme was a gift from Erik Sontheimer (Addgene plasmid # 85679 ; http://n2t.net/addgene:85679 ; RRID:Addgene_85679)
|
| 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 pEJS433-pCSDest2-AcrIIC1Nme 4349 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Mammalian expression of Type II-C anti-CRISPR protein AcrIIC1Nme
from Neisseria meningitidis.
ACCESSION .
VERSION .
KEYWORDS pEJS433-pCSDest2-AcrIIC1Nme
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4349)
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 4349)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4349
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 860..878
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 860..877
/label=SP6
/note="SP6 promoter, forward primer"
promoter complement(1204..1221)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 1226..1360
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1250..1269)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1304..1323
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(1473..1493)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(1473..1491)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1512..1528)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1512..1528)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1525..1547)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1536..1552
/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(1560..1590)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1605..1626
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1743..1760)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1914..2502)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1994..2013)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2673..3533)
/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 3296..3315
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3534..3638)
/gene="bla"
/label=AmpR promoter
rep_origin complement(3664..4119)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3801..3822)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4013..4032
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4197..825
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
ORIGIN
1 aattcaatat ggcggacctg gcactgtgcc aactggggag gggtctactt ggcacggtgc
61 caagtttgag gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc
121 caataatggc ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg
181 gccctatgcc aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat
241 agtattccat atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca
301 tataccatat atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg
361 tctctatata tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg
421 cctcccccat tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc
481 aataggacca cccaccattg acgtcaatgg gatggctcat tgcccattca tatccgttct
541 cacgccccct attgacgtca atgacggtaa atggcccact tggcagtaca tcaatatcta
601 ttaatagtaa cttggcaagt acattactat tggaagtacg ccagggtaca ttggcagtac
661 tcccattgac gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca tccccattga
721 cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg
781 cgtgcctaat gggaggtcta tataagcaat gctcgtttag ggaaccgcca ttctgcctgg
841 ggacgtcgga gcaagcttga tttaggtgac actatagaat acaagctact tgttcttttt
901 gcaggatccc atcgattcga agccgccacc atgaataaaa cttataaaat tggaaaaaat
961 gccgggtatg atggctgcgg tctttgtctt gcggccattt ctgaaaatga agctatcaaa
1021 gttaagtatt tgcgcgacat ttgtcctgat tacgatggcg atgataaagc tgaggattgg
1081 ctgagatggg gaacggacag ccgcgtcaaa gcagccgctc ttgaaatgga gcagtacgca
1141 tatacgtcgg ttggtatggc ctcatgttgg gagtttgttg aactatgact cgagcctcta
1201 gaactatagt gagtcgtatt acgtagatcc agacatgata agatacattg atgagtttgg
1261 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
1321 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
1381 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc
1441 aatgcattgg gcccggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
1501 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
1561 aacatacgag ccgggagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
1621 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
1681 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
1741 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
1801 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
1861 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
1921 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
1981 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
2041 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
2101 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
2161 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
2221 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
2281 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
2341 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
2401 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
2461 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
2521 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
2581 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
2641 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
2701 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
2761 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
2821 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
2881 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
2941 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
3001 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
3061 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
3121 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
3181 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
3241 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
3301 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
3361 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
3421 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
3481 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
3541 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
3601 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
3661 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
3721 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
3781 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
3841 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
3901 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
3961 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
4021 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
4081 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
4141 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag tcgaccgcca
4201 attcaatatg gcgtatatgg actcatgcca attcaatatg gtggatctgg acctgtgcca
4261 attcaatatg gcgtatatgg actcgtgcca attcaatatg gtggatctgg accccagcca
4321 attcaatatg gcggacttgg caccatgcc
//
