| 质粒名称: | pEJS436-pCSDest2-AcrIIC2Nme |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 85712 |
| 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: | 4466bp |
| 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 AcrIIC2Nme |
| 物种来源: | Neisseria meningitidis |
| 插入大小: | 369bp |
| GenBank编号 | WP_042743678.1 |
| 启动子: | CMV |
| 克隆方法: |
Cloning method
|
| 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: |
pEJS436-pCSDest2-AcrIIC2Nme was a gift from Erik Sontheimer (Addgene plasmid # 85712 ; http://n2t.net/addgene:85712 ; RRID:Addgene_85712)
|
| 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 pEJS436-pCSDest2-AcrIIC2Nme 4463 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Mammalian expression of Type II-C anti-CRISPR protein AcrIIC2Nme
from Neisseria meningitidis.
ACCESSION .
VERSION .
KEYWORDS pEJS436-pCSDest2-AcrIIC2Nme
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4463)
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 4463)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4463
/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(1372..1389)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 1394..1528
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1418..1437)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1472..1491
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(1641..1661)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(1641..1659)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1680..1696)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1680..1696)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1693..1715)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1704..1720
/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(1728..1758)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1773..1794
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1911..1928)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2082..2670)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2162..2181)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2841..3701)
/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 3464..3483
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3702..3806)
/gene="bla"
/label=AmpR promoter
rep_origin complement(3832..4287)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3969..3990)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4181..4200
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4365..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 caccatgagc aaaaacaata ttttcaacaa gtatccaaca
1021 attattcacg gcgaagcgcg gggggagaat gacgaatttg tggtgcatac gcgctacccg
1081 cgattcttgg cgcggaaatc ttttgacgac aatttcacgg gcgaaatgcc tgcaaaacct
1141 gttaacgggg aattgggaca aatcggcgaa ccgcgccgcc ttgcttatga ttcacggctt
1201 ggtttgtggc tttctgactt catcatgttg gacaacaaca agccgaaaaa catggaggat
1261 tggcttgggc aattaaaagc cgcctgcgat cgaatcgcgg cggatgattt gatgctgaat
1321 gaagatgcgg cggatttgga gggctgggat gattgactcg agcctctaga actatagtga
1381 gtcgtattac gtagatccag acatgataag atacattgat gagtttggac aaaccacaac
1441 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
1501 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
1561 ggttcagggg gaggtgtggg aggtttttta attcgcggcc gcggcgccaa tgcattgggc
1621 ccggtaccca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
1681 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
1741 gggagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
1801 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
1861 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
1921 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
1981 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
2041 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
2101 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
2161 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
2221 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
2281 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
2341 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
2401 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
2461 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
2521 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
2581 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
2641 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
2701 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
2761 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
2821 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
2881 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
2941 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
3001 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
3061 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
3121 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
3181 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
3241 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
3301 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
3361 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
3421 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
3481 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
3541 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
3601 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
3661 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
3721 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
3781 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
3841 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
3901 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
3961 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
4021 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
4081 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
4141 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
4201 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
4261 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
4321 aagggcgatc ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat tcaatatggc
4381 gtatatggac tcatgccaat tcaatatggt ggatctggac ctgtgccaat tcaatatggc
4441 gtatatggac tcgtgccaat tca
//
