| 質粒名稱: | pRK793 |
|---|---|
| 出品公司: | 愛迪基因 |
| 目錄編號: | 8827 |
| 存儲實驗室: | David Waugh |
|
載體骨架基本信息 |
|
| 載體名稱: | pMal-C2 |
| 載體出品公司: | New England Biolabs |
| 載體抗性: | 氯霉素/氨芐青霉素 |
| 空載體大小: | 6700 bp |
| 完整質粒大小: | -- |
| 載體修飾: | -- |
| 載體類型: | 細菌表達 |
| 篩選標記: | -- |
| 高拷貝/低拷貝: | 低拷貝 |
| 生長條件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名稱: | TEV protease, S219V mutant,tobacco etch virus protease |
| 物種來源: | -- |
| 插入大小: | 750 bp |
| 基因突變: | S219V mutation (improves stability) |
| GenBank編號 | -- |
| 啟動子 | -- |
| 標簽/融合蛋白: |
His（插入件上的N端子）
polyarginine（插入物上的C端）
MBP（帶TEV）（插入件上的N端子）
|
| 克隆方法: | 限制性內切酶 |
| 5' 克隆位點: | SacI(未破壞) |
| 3' 克隆位點: | BamHI (未破壞) |
| 5' 測序引物 | -- |
| 3' 測序引物 | M13-F20 |
|
文獻引用方法 |
|
| 材料和方法部分: | pRK793 was a gift from David Waugh (Addgene plasmid # 8827 ; //n2t.net/addgene:8827 ; RRID:Addgene_8827) |
| 參考文獻部分: | 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 |
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pRK793載體質粒基本信息
pRK793質粒圖譜載體圖譜和pRK793載體序列質粒序列多克隆位點信息
pRK793質粒載體簡介
pRK793質粒序列載體序列
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
//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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