| 质粒名称: | pTargetF |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 62226 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | Trc99a |
| 载体抗性: | 大观霉素 |
| 空载体大小: | 2000 bp |
| 载体修饰: | Replaced Amp with aadA |
| 载体类型: | 细菌表达，CRISPR |
| 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: | sgRNA |
| 物种来源: | Synthetic |
| 启动子 | pij23119 |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | 未知(未破坏) |
| 3' 克隆位点: | 未知 (未破坏) |
| 5' 测序引物 | MoClo-F (agcgaggaagcggaagagcg) |
|
文献引用方法 |
|
| 材料和方法部分: | pTargetF was a gift from Sheng Yang (爱迪基因 plasmid # 62226) |
| 参考文献部分: | Multigene editing in the Escherichia coli genome using the CRISPR-Cas9 system. Jiang Y, Chen B, Duan C, Sun B, Yang J, Yang S. Appl Environ Microbiol. 2015 Jan 30. pii: AEM.04023-14. 10.1128/AEM.04023-14 PubMed 25636838 |
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 pTargetF 2117 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Constitutive expression of sgRNA without donor editing template DNA.
ACCESSION .
VERSION .
KEYWORDS pTargetF
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2117)
AUTHORS Jiang Y, Chen B, Duan C, Sun B, Yang J, Yang S
TITLE Multigene editing in the Escherichia coli genome using the
CRISPR-Cas9 system.
JOURNAL Appl Environ Microbiol. 2015 Jan 30. pii: AEM.04023-14.
PUBMED 25636838
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pTargetF|
FEATURES Location/Qualifiers
source 1..2117
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 98..115
/label=L4440
/note="L4440 vector, forward primer"
promoter 192..226
/label=J23119(SpeI) promoter
/note="bacterial promoter (Registry of Standard Biological
Parts BBa_J23119) modified to end with an SpeI site"
misc_RNA 247..322
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS 508..1299
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 1473..2061
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1962..1981
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
121 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
181 atgctggatc cttgacagct agctcagtcc taggtataat actagtcatc gccgcagcgg
241 tttcaggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
301 aaaagtggca ccgagtcggt gctttttttg aattctctag agtcgacctg cagaagctta
361 gatctattac cctgttatcc ctactcgagt tcatgtgcag ctccataagc aaaaggggat
421 gataagttta tcaccaccga ctatttgcaa cagtgccgtt gatcgtgcta tgatcgactg
481 atgtcatcag cggtggagtg caatgtcatg agggaagcgg tgatcgccga agtatcgact
541 caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta
601 catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg
661 gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg
721 gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt
781 gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa
841 tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg
901 gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag
961 gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg
1021 ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc
1081 cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg
1141 gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat
1201 cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac
1261 tacgtgaaag gcgagatcac caaggtagtc ggcaaataag atgccgctcg ccagtcgatt
1321 ggctgagctc ataagttcct attccgaagt tccgcgaacg cgtaaaggat ctaggtgaag
1381 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
1441 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
1501 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
1561 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
1621 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
1681 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
1741 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
1801 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
1861 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
1921 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
1981 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
2041 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
2101 tgctggcctt ttgctca
//
