| 质粒名称: | phU6-gRNA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 53188 |
| 存储实验室: | Charles Gersbach |
|
载体骨架基本信息 |
|
| 载体名称: | pzDonor |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 3515 bp |
| 载体类型: | 哺乳细胞表达，CRISPR |
| 高拷贝/低拷贝: | 未知 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 启动子 | human U6 |
| 克隆方法: | 限制性内切酶 |
| 3' 测序引物 | M13 Reverse |
|
文献引用方法 |
|
| 材料和方法部分: | phU6-gRNA was a gift from Charles Gersbach (爱迪基因 plasmid # 53188) |
| 参考文献部分: | Multiplex CRISPR/Cas9-based genome engineering from a single lentiviral vector. Kabadi AM, Ousterout DG, Hilton IB, Gersbach CA. Nucleic Acids Res. 2014 Aug 13. pii: gku749. 10.1093/nar/gku749 PubMed 25122746 |
买家导航
phU6-gRNA载体质粒基本信息
phU6-gRNA质粒图谱载体图谱和phU6-gRNA载体序列质粒序列多克隆位点信息
phU6-gRNA质粒载体简介
phU6-gRNA质粒序列载体序列
LOCUS phU6-gRNA 3515 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Expresses the S. pyogenes sgRNA from the human U6 promoter.
ACCESSION .
VERSION .
KEYWORDS phU6-gRNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3515)
AUTHORS Kabadi AM, Ousterout DG, Hilton IB, Gersbach CA
TITLE Multiplex CRISPR/Cas9-based genome engineering from a single
lentiviral vector.
JOURNAL Nucleic Acids Res. 2014 Aug 13. pii: gku749.
PUBMED 25122746
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|phU6-gRNA|
FEATURES Location/Qualifiers
source 1..3515
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(140..161)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 352..371
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 585..607
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 599..616
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 600..616
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 626..645
/label=T7
/note="T7 promoter, forward primer"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 696..936
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 696..716
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 867..886
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 963..1038
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind complement(1094..1114)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(1094..1112)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1133..1149)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1133..1149)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1146..1168)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1157..1173
/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(1181..1211)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1226..1247
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1364..1381)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1535..2123)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1615..1634)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2200..2994)
/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(2311..2330)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 2921..2940
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(3029..3386)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3043..3178
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(3097..3116)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 3343..3363
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter complement(3388..3492)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtacccg
661 tctcacaggc ggatcgatcc aaggtcgggc aggaagaggg cctatttccc atgattcctt
721 catatttgca tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa
781 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg
841 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt
901 tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgggtct tcgagaagac
961 ctgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa
1021 gtggcaccga gtcggtgctt tttttctccg ctgagcgtac tgagacgccg cggtggagct
1081 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
1141 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
1201 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
1261 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
1321 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
1381 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
1441 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
1501 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
1561 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
1621 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
1681 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
1741 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
1801 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
1861 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
1921 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
1981 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
2041 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
2101 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
2161 agtggaacga aaactcacgt taagggattt tggtcatgat cagaagaact cgtcaagaag
2221 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
2281 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg
2341 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc
2401 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg
2461 catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc
2521 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg
2581 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc
2641 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc
2701 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc
2761 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttgcagttc
2821 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag
2881 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag
2941 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa
3001 cgatcctcat cctgtctctt gatcgatctt tgcaaaagcc taggcctcca aaaaagcctc
3061 ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa
3121 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
3181 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc
3241 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc
3301 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
3361 tccacagctg gttctttccg cctcaggact cttccttttt caatattatt gaagcattta
3421 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
3481 aggggttccg cgcacatttc cccgaaaagt gccac
//
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