| 质粒名称: | pCRISPR |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 42875 |
| 存储实验室: | Luciano Marraffini |
|
载体骨架基本信息 |
|
| 载体名称: | pZE21-MCS1 |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 2254 bp |
| 完整质粒大小: | 2433 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达，CRISPR；大肠杆菌 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | CRISPR-BsaI |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pCRISPR was a gift from Luciano Marraffini (Addgene plasmid # 42875 ; http://n2t.net/addgene:42875 ; RRID:Addgene_42875) |
| 参考文献部分: | RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Jiang W, Bikard D, Cox D, Zhang F, Marraffini LA. Nat Biotechnol. 2013 Jan 29. doi: 10.1038/nbt.2508. 10.1038/nbt.2508 PubMed 23360965 |
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 pCRISPR 2707 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION A crRNA expression plasmid for targeting a specific sequence..
ACCESSION .
VERSION .
KEYWORDS pCRISPR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2707)
AUTHORS Jiang W, Bikard D, Cox D, Zhang F, Marraffini LA
TITLE RNA-guided editing of bacterial genomes using CRISPR-Cas systems.
JOURNAL Nat Biotechnol. 2013 Jan 29. doi: 10.1038/nbt.2508.
PUBMED 23360965
REFERENCE 2 (bases 1 to 2707)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..2707
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(15..33)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
CDS 212..1006
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(266..285)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 876..895
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
terminator 1037..1131
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
rep_origin 1219..1807
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1708..1727
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator 1972..2058
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
misc_feature 2088..2219
/label=crRNA leader
/note="crRNA leader sequence for the Streptococcus pyogenes
CRISPR/Cas system"
repeat_region 2220..2255
/label=DR
/note="direct repeat for the Streptococcus pyogenes
CRISPR/Cas system"
repeat_region 2286..2321
/label=DR
/note="direct repeat for the Streptococcus pyogenes
CRISPR/Cas system"
misc_feature 2362..2493
/label=crRNA leader
/note="crRNA leader sequence for the Streptococcus pyogenes
CRISPR/Cas system"
repeat_region 2494..2529
/label=DR
/note="direct repeat for the Streptococcus pyogenes
CRISPR/Cas system"
repeat_region 2560..2595
/label=DR
/note="direct repeat for the Streptococcus pyogenes
CRISPR/Cas system"
promoter complement(2628..2701)
/label=PLtetO-1 promoter
/note="modified phage lambda PL promoter with tet operator
sites (Lutz and Bujard, 1997)"
protein_bind 2658..2676
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2683..2701
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
1 gtgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
61 ggtttcttag acgtcggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg
121 caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc
181 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
241 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
301 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
361 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
421 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
481 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
541 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
601 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
661 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
721 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
781 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
841 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
901 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
961 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
1021 ttcgagagct cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg
1081 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagcact
1141 agtcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
1201 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
1261 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
1321 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
1381 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
1441 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
1501 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
1561 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
1621 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
1681 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
1741 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
1801 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
1861 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
1921 gtgagctgat accgctcgcc gcagccgaac gccctaggtc tagggcggcg gatttgtcct
1981 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg
2041 agcctttcgt tttatttgat gcctctagca cgcgtaccat gggatcctat ttcttaataa
2101 ctaaaaatat ggtataatac tcttaataaa tgcagtaata caggggcttt tcaagactga
2161 agtctagctg agacaaatag tgcgattacg aaatttttta gacaaaaata gtctacgagg
2221 ttttagagct atgctgtttt gaatggtccc aaaactgaga ccagtctcgg aagctcaaag
2281 gtctcgtttt agagctatgc tgttttgaat ggtcccaaaa cttcagcaca ctgagacttg
2341 ttgagttgaa ttcatggatc ctatttctta ataactaaaa atatggtata atactcttaa
2401 taaatgcagt aatacagggg cttttcaaga ctgaagtcta gctgagacaa atagtgcgat
2461 tacgaaattt tttagacaaa aatagtctac gaggttttag agctatgctg ttttgaatgg
2521 tcccaaaact gagaccagtc tcggaagctc aaaggtctcg ttttagagct atgctgtttt
2581 gaatggtccc aaaacttcag cacactgaga cttgttgagt tgaattcggt cagtgcgtcc
2641 tgctgatgtg ctcagtatct ctatcactga tagggatgtc aatctctatc actgataggg
2701 actcgag
//
