| 質粒名稱: | pCRISPR |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 42875 |
| 存儲實驗室: | Luciano Marraffini |
|
載體骨架基本信息 |
|
| 載體名稱: | pZE21-MCS1 |
| 載體出品公司: | -- |
| 載體抗性: | 卡那霉素 |
| 空載體大小: | 2254 bp |
| 完整質粒大小: | 2433 bp |
| 載體修飾: | -- |
| 載體類型: | 細菌表達，CRISPR；大腸桿菌 |
| 篩選標記: | -- |
| 高拷貝/低拷貝: | 高拷貝 |
| 生長條件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名稱: | CRISPR-BsaI |
| 物種來源: | -- |
| 插入大小: | -- |
| 基因突變: | -- |
| GenBank編號 | -- |
| 啟動子 | -- |
| 標簽/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位點: | -- |
| 3' 克隆位點: | -- |
| 5' 測序引物 | -- |
| 3' 測序引物 | -- |
|
文獻引用方法 |
|
| 材料和方法部分: | pCRISPR was a gift from Luciano Marraffini (Addgene plasmid # 42875 ; //n2t.net/addgene:42875 ; RRID:Addgene_42875) |
| 參考文獻部分: | 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 |
買家導航
pCRISPR載體質粒基本信息
pCRISPR質粒圖譜載體圖譜和pCRISPR載體序列質粒序列多克隆位點信息
pCRISPR質粒載體簡介
pCRISPR質粒序列載體序列
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
//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
//