pRK793

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联系方式:I47-825O-882O

相关技术服务:质粒构建    基因合成    质粒大提

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载体基本信息

质粒名称: pRK793 
出品公司: 爱迪基因
目录编号: 8827
存储实验室: David Waugh

载体骨架基本信息
载体名称: pMal-C2
载体出品公司: New England Biolabs
载体抗性: 氯霉素/氨苄青霉素
空载体大小: 6700 bp
完整质粒大小: --
载体修饰: --
载体类型: 细菌表达
筛选标记: --
高拷贝/低拷贝: 低拷贝
生长条件: 37 ℃

插入基因信息
插入基因名称: TEV protease, S219V mutant,tobacco etch virus protease
物种来源: --
插入大小: 750 bp
基因突变: S219V mutation (improves stability)
GenBank编号 --
启动子 --
标签/融合蛋白:
His(插入件上的N端子)
polyarginine(插入物上的C端)
MBP(带TEV)(插入件上的N端子)
克隆方法: 限制性内切酶
5' 克隆位点: SacI(未破坏)
3' 克隆位点: BamHI (未破坏)
5' 测序引物 --
3' 测序引物 M13-F20

文献引用方法
材料和方法部分: pRK793 was a gift from David Waugh (Addgene plasmid # 8827 ; http://n2t.net/addgene:8827 ; RRID:Addgene_8827)
参考文献部分: Tobacco etch virus protease: mechanism of autolysis and rational design of stable mutants with wild-type catalytic proficiency. Kapust RB, Tozser J, Fox JD, Anderson DE, Cherry S, Copeland TD, Waugh DS. Protein Eng. 2001 Dec . 14(12):993-1000. 10.1093/protein/14.12.993 PubMed 11809930

载体图谱质粒图谱和多克隆位点信息

pRK793质粒图谱

载体简介

载体序列

LOCUS       pRK793                3586 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pRK793
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3586)
  AUTHORS   Kapust RB, Tozser J, Fox JD, Anderson DE, Cherry S, Copeland TD, 
            Waugh DS
  TITLE     Tobacco etch virus protease: mechanism of autolysis and rational 
            design of stable mutants with wild-type catalytic proficiency.
  JOURNAL   Protein Eng. 2001 Dec . 14(12):993-1000.
  PUBMED    11809930
REFERENCE   2  (bases 1 to 3586)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3586
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     97..114
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        506..608
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             609..1268
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(675..694)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     tRNA            complement(2561..2637)
                     /gene="argU"
                     /product="E. coli tRNA-Arg recognizing AGA and AGG"
                     /label=argU
     promoter        complement(2881..2909)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      3021..3566
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     3467..3486
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctcacttccc tgttaagtat cttcctggca tcttccagga aatctccgcc ccgttcgtaa
       61 gccatttccg ctcgccgcag tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg
      121 gaatatatcc tgtatcacat attctgctga cgcaccggtg cagccttttt tctcctgcca
      181 catgaagcac ttcactgaca ccctcatcag tgccaacata gtaagccagt atacactccg
      241 ctagcgctga tgtccggcgg tgcttttgcc gttacgcacc accccgtcag tagctgaaca
      301 ggagggacag ctgatagaaa cagaagccac tggagcacct caaaaacacc atcatacact
      361 aaatcagtaa gttggcagca tcacccgacg cactttgcgc cgaataaata cctgtgacgg
      421 aagatcactt cgcagaataa ataaatcctg gtgtccctgt tgataccggg aagccctggg
      481 ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag aggttccaac tttcaccata
      541 atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg
      601 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
      661 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
      721 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
      781 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga
      841 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
      901 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
      961 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
     1021 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
     1081 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
     1141 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     1201 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     1261 gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggttgc tacgcctgaa
     1321 taagtgataa taagcggatg aatggcagaa attcgaaagc aaattcgacc cggtcgtcgg
     1381 ttcagggcag ggtcgttaaa tagccgctta tgtctattgc tggtttaccg gtttattgac
     1441 taccggaagc agtgtgaccg tgtgcttctc aaatgcctga ggccagtttg ctcaggctct
     1501 ccccgtggag gtaataattg acgatatgat catttattct gcctcccaga gcctgataaa
     1561 aacggttagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     1621 gcgatgcaga tccggaacat aatggtgcag ggcgcttgtt tcggcgtggg tatggtggca
     1681 ggccccgtgg ccgggggact gttgggcgct gccggcacct gtcctacgag ttgcatgata
     1741 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagcta
     1801 ccggcagcgg tgcggactgt tgtaactcag aataagaaat gaggccgctc atggcgttga
     1861 ctctcagtca tagtatcgtg gtatcaccgg ttggttccac tctctgttgc gggcaacttc
     1921 agcagcacgt aggggacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca
     1981 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc
     2041 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca
     2101 acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gactcgaggg
     2161 catccgatca acgctttctg gtgaatggtg cgggaggcga gacttgaact cgcacaccta
     2221 gcggcgccag aacctaaatc tggtgcgtct accaatttcg ccactcccgc aaaaaaagat
     2281 ggtggctacg acgggattct ctagatcatc atgtttattg cgtggtttat aagtattgtt
     2341 attgcacact caaggccctt tagctcagtg gttagagcag gcgactcata atcgcttggt
     2401 cgctggttca agtccagcaa gggccaccaa atcattattt acgtatcagg aagagttaag
     2461 ggtacaaggg gtactcgatg gcggtctata tttcatgact aacgccattt caaggccatg
     2521 gctggaggat ataaagaagg cgtaactgat tgaattgtaa tggcgcgccc tgcaggattc
     2581 gaacctgcgg cccacgactt agaaggtcgt tgctctatcc aactgagcta agggcgcgtt
     2641 gataccgcaa tgcggtgtaa tcgcgtgaat tatacggtca acccttgctg agtcaatggc
     2701 ttttgatctg gttgctgaac aagtgaacga ccgcgtctga ttactagttt gcccgggccc
     2761 ttgtcgtcgt caattgtaca gcatccaggg tgacggtgcc gaggatgacg atgagcgcat
     2821 tgttagattt catacacggt gcctgactgc gttagcaatt taactgtgat aaactaccgc
     2881 attaaagctt atcgatgata agctgtcaaa catgagaatt acaacttata tcgtatgggg
     2941 ctgacttcag gtgctacatt tgaagagata aattgcactg aaatctagaa atattttatc
     3001 tgattaataa gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa
     3061 aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc tctgagctac caactctttg
     3121 aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct
     3181 taaccggcgc atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag
     3241 tggtgctttt gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3301 gcggtcggac tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc
     3361 ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg
     3421 taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc gccaggggga aacgcctggt
     3481 atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct
     3541 tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccct
//

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