| 质粒名称: | pVAX_CBI_AcrIIA5_D4276 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 136664 |
| 存储实验室: | Yannick Doyon |
|
载体骨架基本信息 |
|
| 载体名称: | pVAX_CBI |
| 完整质粒大小: | 3936bp |
| 载体类型: | 哺乳细胞表达、Synthetic Biology |
| 载体抗性: | Kanamycin |
| 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: | AcrIIA5_D4276 / |
| 物种来源: | H. sapiens (human), Synthetic |
| 启动子: | CMV |
| 标签/融合蛋白: | No tag, No NLS |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | TAT GGT TGG GAT AAG GCT GG |
|
文献引用方法 |
|
| 材料和方法部分: |
pVAX_CBI_AcrIIA5_D4276 was a gift from Yannick Doyon (Addgene plasmid # 136664 ; http://n2t.net/addgene:136664 ; RRID:Addgene_136664)
|
| 参考文献部分: | Widespread anti-CRISPR proteins in virulent bacteriophages inhibit a range of Cas9 proteins. Hynes AP, Rousseau GM, Agudelo D, Goulet A, Amigues B, Loehr J, Romero DA, Fremaux C, Horvath P, Doyon Y, Cambillau C, Moineau S. Nat Commun. 2018 Jul 25;9(1):2919. doi: 10.1038/s41467-018-05092-w. 10.1038/s41467-018-05092-w PubMed |
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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 pVAX_CBI_AcrIIA5_D4276 3936 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Expresses human codon-optimized AcrIIA5 D4276 in mammalian cells.
ORF flanked by NotI and XhoI restriction sites.
ACCESSION .
VERSION .
KEYWORDS pVAX_CBI_AcrIIA5_D4276
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3936)
AUTHORS Hynes AP, Rousseau GM, Agudelo D, Goulet A, Amigues B, Loehr J,
Romero DA, Fremaux C, Horvath P, Doyon Y, Cambillau C, Moineau S
TITLE Widespread anti-CRISPR proteins in virulent bacteriophages inhibit a
range of Cas9 proteins.
JOURNAL Nat Commun. 2018 Jul 25;9(1):2919. doi: 10.1038/s41467-018-05092-w.
PUBMED 30046034
REFERENCE 2 (bases 1 to 3936)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3936
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 93..396
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 397..600
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 551..571
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 597..621
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 1214..1261
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
regulatory 1289..1298
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind complement(1758..1775)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1764..1988
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
CDS 2161..2955
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2215..2234)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 2825..2844
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
rep_origin 3280..3868
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3769..3788
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gcttcgcgat gtacgggcca gatatacgcg tctagttatt aatagtaatc aattacgggg
61 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
121 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
181 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
241 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
301 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
361 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
421 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
481 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
541 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
601 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
661 agacaccggg accgatccag cctccgcgga ttcgaatccc ggccgggaac ggtgcattgg
721 aacgcggatt ccccgtgcca agagtgacgt aagtaccgcc tatagagtct ataggcccac
781 aaaaaatgct ttcttctttt aatatacttt tttgtttatc ttatttctaa tactttccct
841 aatctctttc tttcagggca ataatgatac aatgtatcat gcctctttgc accattctaa
901 agaataacag tgataatttc tgggttaagg caatagcaat atttctgcat ataaatattt
961 ctgcatataa attgtaactg atgtaagagg tttcatattg ctaatagcag ctacaatcca
1021 gctaccattc tgcttttatt ttatggttgg gataaggctg gattattctg agtccaagct
1081 aggccctttt gctaatcatg ttcatacctc ttatcttcct cccacagctc ctgggcaacg
1141 tgctggtctg tgtgctggcc catcactttg gcaaagaatt gggattcgaa catcgattga
1201 attcctgtac aagaaaaggc cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa
1261 gggcggaggc tctggcggcg gaagcgcggc cgccatggcc tatggtaaaa gtaggtataa
1321 ctcctatcgc aaacgctctt tcaacaggtc taacaaacaa aggagagaat acgcacagga
1381 aatggaccgc cttgaaaagg catttgagaa cctcgacgga tggtacttga gtagcatgaa
1441 agattctgcg tataaggatt tcggtaagta cgaaatccga ctttcaaatc actcagccga
1501 caataaatat catgacctgg agaacggccg cttgatcgtg aatatcaaag caagcaaact
1561 taactttgtc gatattatcg agaataaact cgacaagatc atcgaaaaaa ttgataagct
1621 ggaccttgat aaatatcgct tcatcaatgc gaccaacctg gagcacgaca ttaagtgcta
1681 ctataaaggg tttaagacta agaaagaggt tatctgataa ctcgagtcta gagggcccgt
1741 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
1801 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
1861 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
1921 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
1981 ctctatggct tctactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg
2041 gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca
2101 aggatctgat ggcgcagggg atcaagctct gatcaagaga caggatgagg atcgtttcgc
2161 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
2221 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
2281 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
2341 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
2401 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
2461 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
2521 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
2581 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
2641 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
2701 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
2761 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
2821 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
2881 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
2941 gacgagttct tctgaattat taacgcttac aatttcctga tgcggtattt tctccttacg
3001 catctgtgcg gtatttcaca ccgcatcagg tggcactttt cggggaaatg tgcgcggaac
3061 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
3121 ctgataaatg cttcaataat agcacgtgct aaaacttcat ttttaattta aaaggatcta
3181 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
3241 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
3301 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
3361 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
3421 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
3481 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
3541 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
3601 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
3661 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
3721 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
3781 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
3841 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
3901 ggccttttgc tggccttttg ctcacatgtt cttgct
//
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