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pRSFDuet1載體基本信息
EMD Biosciences (Novagen) |
pRSFDuet1, pRSFDuet 1 |
大腸桿菌蛋白雙表達載體,有兩個多克隆位點 |
高 |
多克隆位點,限制性內切酶 |
3829 bp |
T7 |
T7: 5'-TAATACGACTCACTATAGGG-3'; |
T7t |
T7t: 5'-TGCTAGTTATTGCTCAGCGG-3' |
N-His, C-S |
Kanamycin |
pRSFDuet-1 carries the RSF1030 replicon, lacI gene and kanamycin resistance gene. |
71341-3 |
瞬時表達 Transient |
組成型 Constitutive |
非病毒 |
pRSFDuet1載體質粒圖譜和多克隆位點信息
pRSFDuet1載體簡介
pRSFDuet-1 is designed for the coexpression of two target ORFs. The vector contains two multiple cloning sites (MCS), each of which is preceded by a T7lac promoter and ribosome binding site (rbs). The vector also carries the RSF1030-derived RSF replicon, lacI gene and kanamycin resistance gene (KnR). This vector can be used in combination with pACYCDuet™-1 (Cat. No. 71147-3), pCDFDuet™-1 (Cat. Np. 71340-3), and pETDuet™-1 (Cat. No. 71146-3) in an appropriate host strain for the coexpression of 4 to 8 target proteins. ORFs inserted into MCS1 can be sequenced using the ACYCDuetUP1 Primer (Cat. No. 71178-3) and DuetDOWN1 Primer (Cat. No. 71179-3). ORFs inserted into MCS2 can be sequenced using the DuetUP2 Primer (Cat. No. 71180-3) and T7 Terminator Primer (Cat. No. 69337-3). Unique sites are shown on the circle map.
pRSFDuet1載體序列
DEFINITION pRSFDuet-1
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTOAUTHORAD1|Novagen Inc.|
FEATURES Location/Qualifiers
source 1..3829
/organism="pRSFDuet-1"
/mol_type="other DNA"
misc_feature 2..29
/label="lacO"
promoter 214..232
/label="T7_promoter"
misc_feature 232..259
/label="lacO"
misc_feature complement(448..466)
/label="T7_Terminal_primer"
gene complement(739..1554)
/label="KanR2"
/gene="KanR2"
CDS complement(739..1554)
/label="ORF frame 2"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
promoter complement(1596..1624)
/label="AmpR_promoter"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
CDS complement(2607..3566)
/label="ORF frame 3"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
CDS complement(3439..158)
/label="ORF frame 3"
/translation="MRPQACRPAGAPSSNSDPGCGDDGDGCCPWYISLLKLNKIISTG
ELLSAHNSPIVSRINFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECET
SNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTV
VADWRCHLQSGPARAVANCRGD*"
promoter 3813..2
/label="T7_promoter"
/translation="MRPQACRPAGAPSSNSDPGCGDDGDGCCPWYISLLKLNKIISTG
ELLSAHNSPIVSRINFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECET
SNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTV
VADWRCHLQSGPARAVANCRGD*"
ORIGIN
1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag
181 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt
241 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata
301 tggcagatct caattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt
361 ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg
421 cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg
481 cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg
541 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
601 aacgaccctg ccctgaaccg acgacaagct gacgaccggg tctccgcaag tggcactttt
661 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
721 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
781 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
841 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
901 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
961 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
1021 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
1081 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cggtcgctgt taaaaggaca
1141 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
1201 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
1261 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
1321 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
1381 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
1441 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
1501 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt
1561 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
1621 tgaatgtatt tagaaaaata aacaaatagg catgcagcgc tcttccgctt cctcgctcac
1681 tgactcgcta cgctcggtcg ttcgactgcg gcgagcggtg tcagctcact caaaagcggt
1741 aatacggtta tccacagaat caggggataa agccggaaag aacatgtgag caaaaagcaa
1801 agcaccggaa gaagccaacg ccgcaggcgt ttttccatag gctccgcccc cctgacgagc
1861 atcacaaaaa tcgacgctca agccagaggt ggcgaaaccc gacaggacta taaagatacc
1921 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
1981 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgtt
2041 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
2101 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
2161 acgacttatc gccactggca gcagccattg gtaactgatt tagaggactt tgtcttgaag
2221 ttatgcacct gttaaggcta aactgaaaga acagattttg gtgagtgcgg tcctccaacc
2281 cacttacctt ggttcaaaga gttggtagct cagcgaacct tgagaaaacc accgttggta
2341 gcggtggttt ttctttattt atgagatgat gaatcaatcg gtctatcaag tcaacgaaca
2401 gctattccgt tactctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
2461 agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg
2521 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
2581 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
2641 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
2701 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
27
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