| 质粒名称: | pTarget12f1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 171183 |
| 存储实验室: | Kohsuke Honda |
|
载体骨架基本信息 |
|
| 载体名称: | pTargetF |
| 载体出品公司: | -- |
| 空载体大小: | 2014 bp |
| 完整质粒大小: | 2217 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达，CRISPR |
| 筛选标记: | -- |
| 载体抗性: | 大观霉素，50μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | CRISPR/Cas12f1系统中的sgRNA支架 |
| 物种来源: | Synthetic |
| 插入大小: | 951 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | J23119（SpeI）启动子 |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | SpeI (未破坏) |
| 3’克隆位点: | EcoRI (未破坏) |
| 5’测序引物 | CTCCTTACGCATCTGTGCG |
| 3’测序引物 | GATCAACGGCACTGTTGC |
|
文献引用方法 |
|
| 材料和方法部分: | pTarget12f1 was a gift from Kohsuke Honda (Addgene plasmid # 171183 ; http://n2t.net/addgene:171183 ; RRID:Addgene_171183) |
| 参考文献部分: | Genome editing by miniature CRISPR/Cas12f1 enzyme in Escherichia coli. Okano K, Sato Y, Hizume T, Honda K. J Biosci Bioeng. 2021 Aug;132(2):120-124. doi: 10.1016/j.jbiosc.2021.04.009. Epub 2021 May 19. 10.1016/j.jbiosc.2021.04.009 PubMed 34023220 |
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 pTarget12f1 2243 bp ds-DNA circular SYN 29-JUN-2021
DEFINITION Constitutive expression of sgRNA without donor DNA.
ACCESSION .
VERSION .
KEYWORDS pTarget12f1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2243)
TITLE Genome editing by miniature CRISPR/Cas12f1 enzyme in Escherichia
coli
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 2243)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jun 29, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..2243
/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"
CDS 634..1425
/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 1599..2187
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2088..2107
/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 actagtcttc actgataaag
241 tggagaaccg cttcaccaaa agctgtccct taggggatta gaacttgagt gaaggtgggc
301 tgcttgcatc agcctaatgt cgagaagtgc tttcttcgga aagtaaccct cgaaacaaat
361 tcatttttcc tctccaattc tgcacaagaa agttgcagaa cccgaataga cgaatgaagg
421 aatgcaactg caccgtgtct agctagcttt tttttgaatt ctctagagtc gacctgcaga
481 agcttagatc tattaccctg ttatccctac tcgagttcat gtgcagctcc ataagcaaaa
541 ggggatgata agtttatcac caccgactat ttgcaacagt gccgttgatc gtgctatgat
601 cgactgatgt catcagcggt ggagtgcaat gtcatgaggg aagcggtgat cgccgaagta
661 tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg
721 gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat
781 ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac
841 cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc
901 attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt
961 ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt
1021 gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg
1081 gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc
1141 ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg
1201 ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc
1261 gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag
1321 gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt
1381 gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataagatgc cgctcgccag
1441 tcgattggct gagctcataa gttcctattc cgaagttccg cgaacgcgta aaggatctag
1501 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
1561 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
1621 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
1681 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
1741 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
1801 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
1861 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
1921 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
1981 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
2041 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
2101 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
2161 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
2221 gccttttgct ggccttttgc tca
//
