| 质粒名称: | pEcgRNA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 166581 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pTargetF |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 壮观霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | ccdB Survival |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | ccdB |
| 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: | Ligation Independent 克隆 |
| 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: | pEcgRNA was a gift from Sheng Yang (Addgene plasmid # 166581 ; http://n2t.net/addgene:166581 ; RRID:Addgene_166581) |
| 参考文献部分: | A modified pCas/pTargetF system for CRISPR-Cas9-assisted genome editing in Escherichia coli. Li , Q i, Sun, Bingbing, Chen, Jun, Zhang, Yiwen, Jiang, Y u Jiang, Yang, Sheng. Acta Biochimica et Biophysica Sinica, 2021 10.1093/abbs/gmab036 |
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 pEcgRNA 3134 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION CRISPR-Cas9-assisted genome editing in Escherichia coli.
ACCESSION .
VERSION .
KEYWORDS pEcgRNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3134)
AUTHORS Li , Q i, Sun, Bingbing, Chen, Jun, Zhang, Yiwen, Jiang, Y u Jiang,
Yang, Sheng
TITLE A modified pCas/pTargetF system for CRISPR-Cas9-assisted genome
editing in Escherichia coli
JOURNAL Acta Biochimica et Biophysica Sinica, 2021
REFERENCE 2 (bases 1 to 3134)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3134
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 45..62
/label=L4440
/note="L4440 vector, forward primer"
promoter 139..173
/label=J23119(SpeI) promoter
/note="bacterial promoter (Registry of Standard Biological
Parts BBa_J23119) modified to end with an SpeI site"
CDS complement(560..865)
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind complement(662..681)
/label=ccdB-fwd
/note="ccdB gene, forward primer"
misc_RNA 1211..1286
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS 1472..2263
/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 2437..3025
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2926..2945
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
61 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
121 accgcatatg ctggatcctt gacagctagc tcagtcctag gtataatact agttgagacc
181 cagcagtgaa agataaatga tctgagaccc aggtcgaccg acagccttcc aaatgttctt
241 ctcaaacgga atcgtcgtat ccagcctact cgctattgtc ctcaatgccg tattaaatca
301 taaaaagaaa taagaaaaag aggtgcgagc ctcttttttg tgtgacaaaa taaaaacatc
361 tacctattca tatacgctag tgtcatagtc ctgaaaatca tctgcatcaa gaacaatttc
421 acaactctta tacttttctc ttacaagtcg ttcggcttca tctggatttt cagcctctat
481 acttactaaa cgtgataaag tttctgtaat ttctactgta tcgacctgca gactggctgt
541 gtataaggga gcctgacatt tatattcccc agaacatcag gttaatggcg tttttgatgt
601 cattttcgcg gtggctgaga tcagccactt cttccccgat aacggagacc ggcacactgg
661 ccatatcggt ggtcatcatg cgccagcttt catccccgat atgcaccacc gggtaaagtt
721 cacgggagac tttatctgac agcagacgtg cactggccag ggggatcacc atccgtcgcc
781 cgggcgtgtc aataatatca ctctgtacat ccacaaacag acgataacgg ctctctcttt
841 tataggtgta aaccttaaac tgcatttcac cagcccctgt tctcgtcagc aaaagagccg
901 ttcatttcaa taaaccgggc gacctcagcc atcccttcct gattttccgc tttccagcgt
961 tcggcacgca gacgacgggc ttcattctgc atggttgtgc ttaccagacc ggagatattg
1021 acatcatata tgccttgagc aactgatagc tgtcgctgtc aactgtcact gtaatacgct
1081 gcttcatagc atacctcttt ttgacatact tcgggtatac atatcagtat atattcttat
1141 accgcaaaaa tcagcgcgca aatacgcata ctgttatctg gcttttagta agccggatcc
1201 ctgggtctcg gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac
1261 ttgaaaaagt ggcaccgagt cggtgctttt tttgaattct ctagagtcga cctgcagaag
1321 cttagatcta ttaccctgtt atccctactc gagttcatgt gcagctccat aagcaaaagg
1381 ggatgataag tttatcacca ccgactattt gcaacagtgc cgttgatcgt gctatgatcg
1441 actgatgtca tcagcggtgg agtgcaatgt catgagggaa gcggtgatcg ccgaagtatc
1501 gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc
1561 cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt
1621 gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct
1681 tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat
1741 tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg
1801 agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga
1861 tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc
1921 ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt
1981 aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt
2041 gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga
2101 ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc
2161 ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt
2221 ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taagatgccg ctcgccagtc
2281 gattggctga gctcataagt tcctattccg aagttccgcg aacgcgtaaa ggatctaggt
2341 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
2401 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
2461 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
2521 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
2581 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
2641 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
2701 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
2761 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
2821 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
2881 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
2941 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
3001 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
3061 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
3121 ccgtattacc gcct
//
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