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pCDFDuet-1 构建费用600元/个起

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基本信息

别名

pCDFDuet-1; pCDFDuet1; pCDFDuet 1

体类型

大肠杆菌蛋白双表达载体,有两个多克隆位点(双启动子)

载体抗性

链霉素 streptomycin

载体标签

N-His, C-S

筛选标记

/

启动子

T7lac

表达水平

低拷贝

载体大小

3871bp

克隆方法

多克隆位点,限制性内切酶

5' 测序引物

DUETF1: TGGTGTCCGGGATCTCGA
DUETF2: TTGTACACGGCCGCATAATC

3' 测序引物

DUETR1: 5'-GATTATGCGGCCGTGTACAA-3' 
T7 ter: TGCTAGTTATTGCTCAGCGG

组成型

诱导型

备注

pCDFDuet-1 is designed for the coexpression of two target genes. pCDFDuet-1 carries the CloDF13 replicon, lacI gene and

质粒图谱和多克隆位点信息

注意:

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如要使用本公司保存的母源载体时,本公司不能保证所有母源载体完全符合客户预期,也不承担由此产生的任何责任。

 

 

 

载体描述

pCDFDuet™-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 CloDF13-derived CDF replicon, lacI gene and streptomycin/spectinomycin resistance gene (SmR). This vector can be used in combination with pACYCDuet™-1 (Cat. No. 71147-3), pRSFDuet™-1 (Cat. No. 71341-3), 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.

 

载体序列

LOCUS       pCDFDuet-1                3781 bp ds-DNA     circular SYN 23-MAR-2019

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    pCDFDuet-1

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 3781)

  AUTHORS   BT-Lab

  TITLE     Direct Submission

  JOURNAL   Exported Saturday, Mar 23, 2019 from SnapGene Viewer 4.3.4

FEATURES             Location/Qualifiers

     source          1..3781

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     protein_bind    3..27

                     /label=lac operator

                     /bound_moiety="lac repressor encoded by lacI"

                     /note="The lac repressor binds to the lac operator to

                     inhibit transcription in E. coli. This inhibition can be

                     relieved by adding lactose or

                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."

     CDS             83..100

                     /codon_start=1

                     /product="6xHis affinity tag"

                     /label=6xHis

                     /translation="HHHHHH"

     misc_feature    106..195

                     /label=MCS1

                     /note="multiple cloning site"

     promoter        214..232

                     /label=T7 promoter

                     /note="promoter for bacteriophage T7 RNA polymerase"

     protein_bind    233..257

                     /label=lac operator

                     /bound_moiety="lac repressor encoded by lacI"

                     /note="The lac repressor binds to the lac operator to

                     inhibit transcription in E. coli. This inhibition can be

                     relieved by adding lactose or

                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."

     misc_feature    297..359

                     /label=MCS2

                     /note="multiple cloning site"

     CDS             366..410

                     /codon_start=1

                     /product="affinity and epitope tag derived from pancreatic

                     ribonuclease A"

                     /label=S-Tag

                     /translation="KETAAAKFERQHMDS"

     terminator      462..509

                     /label=T7 terminator

                     /note="transcription terminator for bacteriophage T7 RNA

                     polymerase"

     CDS             complement(680..1471)

                     /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"

     promoter        complement(1472..1563)

                     /gene="bla"

                     /label=AmpR promoter

     rep_origin      complement(1611..2349)

                     /direction=LEFT

                     /label=CloDF13 ori

                     /note="Plasmids containing the CloDF13 (CDF) origin of

                     replication can be propagated in E. coli cells that contain

                     additional plasmids with compatible origins."

     CDS             complement(2559..3641)

                     /codon_start=1

                     /gene="lacI"

                     /product="lac repressor"

                     /label=lacI

                     /note="The lac repressor binds to the lac operator to

                     inhibit transcription in E. coli. This inhibition can be

                     relieved by adding lactose or

                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."

                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL

                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV

                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH

                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA

                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC

                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR

                     ALADSLMQLARQVSRLESGQ"

     promoter        complement(3642..3719)

                     /gene="lacI"

                     /label=lacI promoter

     misc_feature    join(3765..3781,1..2)

                     /label=T7 promoter

                     /note="promoter for bacteriophage T7 RNA polymerase"

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 acgaccgggt catcgtggcc ggatcttgcg gcccctcggc

      661 ttgaacgaat tgttagacat tatttgccga ctaccttggt gatctcgcct ttcacgtagt

      721 ggacaaattc ttccaactga tctgcgcgcg aggccaagcg atcttcttct tgtccaagat

      781 aagcctgtct agcttcaagt atgacgggct gatactgggc cggcaggcgc tccattgccc

      841 agtcggcagc gacatccttc ggcgcgattt tgccggttac tgcgctgtac caaatgcggg

      901 acaacgtaag cactacattt cgctcatcgc cagcccagtc gggcggcgag ttccatagcg

      961 ttaaggtttc atttagcgcc tcaaatagat cctgttcagg aaccggatca aagagttcct

     1021 ccgccgctgg acctaccaag gcaacgctat gttctcttgc ttttgtcagc aagatagcca

     1081 gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag aatgtcattg cgctgccatt

     1141 ctccaaattg cagttcgcgc ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa

     1201 caatggtgac ttctacagcg cggagaatct cgctctctcc aggggaagcc gaagtttcca

     1261 aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag ccttacggtc accgtaacca

     1321 gcaaatcaat atcactgtgt ggcttcaggc cgccatccac tgcggagccg tacaaatgta

     1381 cggccagcaa cgtcggttcg agatggcgct cgatgacgcc aactacctct gatagttgag

     1441 tcgatacttc ggcgatcacc gcttccctca tactcttcct ttttcaatat tattgaagca

     1501 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac

     1561 aaatagctag ctcactcggt cgctacgctc cgggcgtgag actgcggcgg gcgctgcgga

     1621 cacatacaaa gttacccaca gattccgtgg ataagcaggg gactaacatg tgaggcaaaa

     1681 cagcagggcc gcgccggtgg cgtttttcca taggctccgc cctcctgcca gagttcacat

     1741 aaacagacgc ttttccggtg catctgtggg agccgtgagg ctcaaccatg aatctgacag

     1801 tacgggcgaa acccgacagg acttaaagat ccccaccgtt tccggcgggt cgctccctct

     1861 tgcgctctcc tgttccgacc ctgccgttta ccggatacct gttccgcctt tctcccttac

     1921 gggaagtgtg gcgctttctc atagctcaca cactggtatc tcggctcggt gtaggtcgtt

     1981 cgctccaagc tgggctgtaa gcaagaactc cccgttcagc ccgactgctg cgccttatcc

     2041 ggtaactgtt cacttgagtc caacccggaa aagcacggta aaacgccact ggcagcagcc

     2101 attggtaact gggagttcgc agaggatttg tttagctaaa cacgcggttg ctcttgaagt

     2161 gtgcgccaaa gtccggctac actggaagga cagatttggt tgctgtgctc tgcgaaagcc

     2221 agttaccacg gttaagcagt tccccaactg acttaacctt cgatcaaacc acctccccag

     2281 gtggtttttt cgtttacagg gcaaaagatt acgcgcagaa aaaaaggatc tcaagaagat

     2341 cctttgatct tttctactga accgctctag atttcagtgc aatttatctc ttcaaatgta

     2401 gcacctgaag tcagccccat acgatataag ttgtaattct catgttagtc atgccccgcg

     2461 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg

     2521 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg

     2581 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc

     2641 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc

     2701 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg

     2761 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg

     2821 tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt aatggcgcgc

     2881 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca

     2941 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc

     3001 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc

     3061 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc

     3121 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt

     3181 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca

     3241 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga

     3301 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc

     3361 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg

     3421 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt

     3481 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc

     3541 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca

     3601 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga

     3661 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc

     3721 gggatctcga cgctctccct tatgcgactc ctgcattagg aaattaatac gactcactat

     3781 a

//