| 质粒名称: | pCMV-T7-hAcrVA3.1-NLS(sv40) (RTW2624) |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 115139 |
| 存储实验室: | Joseph Bondy-Denomy, Keith Joung, Benjamin Kleinstiver |
|
载体骨架基本信息 |
|
| 载体名称: | JDS246 (Plasmid #43861) |
| 载体出品公司: | Joung lab (Addgene Plasmid #43861) |
| 载体类型: | 哺乳细胞表达 |
| 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: | human codon optimized AcrVA3.1 anti-CRISPR protein / RTW2624 |
| 物种来源: | Synthetic |
| 标签/融合蛋白: | NLS(SV40) (C terminal on insert) |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | CMV-F |
| 3’测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV-T7-hAcrVA3.1-NLS(sv40) (RTW2624) was a gift from Joseph Bondy-Denomy & Keith Joung & Benjamin Kleinstiver (Addgene plasmid # 115139 ; http://n2t.net/addgene:115139 ; RRID:Addgene_115139) |
| 参考文献部分: | Discovery of widespread Type I and Type V CRISPR-Cas inhibitors. Marino ND, Zhang JY, Borges AL, Sousa AA, Leon LM, Rauch BJ, Walton RT, Berry JD, Joung JK, Kleinstiver BP, Bondy-Denomy J. Science. 2018 Sep 6. pii: science.aau5174. doi: 10.1126/science.aau5174. 10.1126/science.aau5174 PubMed 30190308 |
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
 (RTW2624)质粒图谱.png)
载体简介
载体序列
LOCUS pCMV-T7-hAcrVA3.1-NLS(sv40) (RTW2624) 3951 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION CMV-T7 promoter expression plasmid for human codon optimized
AcrVA3.1 anti-CRISPR protein with C-terminal NLS .
ACCESSION .
VERSION .
KEYWORDS pCMV-T7-hAcrVA3.1-NLS(sv40) (RTW2624)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3951)
AUTHORS Marino ND, Zhang JY, Borges AL, Sousa AA, Leon LM, Rauch BJ, Walton
RT, Berry JD, Joung JK, Kleinstiver BP, Bondy-Denomy J
TITLE Discovery of widespread Type I and Type V CRISPR-Cas inhibitors.
JOURNAL Science. 2018 Sep 6. pii: science.aau5174. doi:
10.1126/science.aau5174.
PUBMED 30190308
REFERENCE 2 (bases 1 to 3951)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3951
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 529..549
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 558..575
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(598..615)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 604..828
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(899..915)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(899..915)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(912..934)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 923..939
/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(947..977)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 992..1013
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1130..1147)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1301..1889)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1381..1400)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2060..2920)
/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 2683..2702
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2921..3025)
/gene="bla"
/label=AmpR promoter
primer_bind complement(3104..3123)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 3295..3674
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3675..3878
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 3829..3849
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 3920..3939
/label=T7
/note="T7 promoter, forward primer"
promoter 3920..3938
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 3946..4
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
ORIGIN
1 atgggcatag cgcatcagaa gaaccggaga gcggattggg aaagcgtaga ctggactaaa
61 cataatgatg agattgccca actgctgagc agacatcctg atagtgtagc aaagatgaga
121 acgaagttcg gcgctcaggg aatggctaaa cggaagccga ggcgcaaata taaggtcaca
181 cgaaaggctg tcccgcctcc tcatacgcag gagctcgcaa cagccgctgc aaaaatctcc
241 ccaaagagtg gacgctacga aaccaacgtg aacgctaaaa gatggctcat catatccccg
301 agtggtcaga ggttcgagtt ctccaacctc cagcatttcg tacgcaatca cccggaattg
361 tttgccaagg cggatacggt ttggaagcgc caggggggaa agaggggtac ggggggcgaa
421 tattgcaacg cgagcaatgg acttgctcag gccgccaggt tgaacatagg gtggaagggg
481 tggcaagcga agatcattaa aggcggatcc ggcggtggag gcagtgggcc caagaagaag
541 aggaaagtct gaccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct
601 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
661 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
721 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
781 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
841 aaagaaccag ctggggctcg ataccgtcga cctctagcta gagcttggcg taatcatggt
901 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
961 gaagcataaa gtgtaaagcc tagggtgcct aatgagtgag ctaactcaca ttaattgcgt
1021 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
1081 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
1141 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
1201 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
1261 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
1321 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
1381 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
1441 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
1501 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
1561 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
1621 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
1681 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
1741 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
1801 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
1861 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
1921 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
1981 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
2041 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
2101 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
2161 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
2221 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
2281 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
2341 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
2401 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
2461 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
2521 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
2581 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
2641 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
2701 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
2761 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
2821 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
2881 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
2941 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
3001 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
3061 gatcgggaga tcgatctccc gatcccctag ggtcgactct cagtacaatc tgctctgatg
3121 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
3181 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
3241 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
3301 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
3361 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
3421 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
3481 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
3541 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
3601 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
3661 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
3721 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
3781 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
3841 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
3901 gctagagatc cgcggccgct aatacgactc actataggga gagccgccac c
//
