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pFastBacDual

pFastBacDual

pFastBacDual

pFastBacDual

 



编号

载体名称

北京华越洋VECT75434

pFastBacDual

 

pFastBacDual载体基本信息:

复制子:

pUC ori,F1 ori

终止子:

SV40 poly(A) signal

质粒分类:

广宿主系列,昆虫细胞载体

质粒大小:

5238bp

原核抗性:

Amp

筛选标记:

Gen

克隆菌株:

DH5α

培养条件:

37℃,有氧 LB

表达宿主:

昆虫细胞

诱导方式:

无须诱导,瞬时表达

5'测序引物:

pFastbac-F: TATTCCGGATTATTCATACC

3'测序引物:

pFastBac-R: ACAAATGTGGTATGGCTGA

 

pFastBacDual载体图谱:


pFastBacDual载体简介:

 pFastBac Dual昆虫细胞表达载体包含有两个启动子,能够同时表达两个外源基因,一个表达基因是polyhedrin (PH)启动子控制,另一个外源基因是由p10启动子控制。
pFastBac™Dual 载体的特点是在单个载体上有两个启动子,用于在使用 Bac-to-Bac® 杆状病毒表达系统(Bac-to-Bac® Baculovirus Expression System) 时在昆虫细胞中同时表达两个蛋白质。该载体具有两个强启动子,多角体蛋白启动子和p10 启动子,因此可实现高水平表达。包含 CAT和β-葡萄糖醛酸酶 (gus) 的表达对照。
   The Bac-to-Bac® Vector Kit contains a pFastBac™ 1 vector, as well an expression control vector, intended for use as part of the Bac-to-Bac® Baculovirus Expression System , which enables the efficent production of recombinant baculovirus for expression testing in insect cells. The Bac-to-Bac® System relies on generation of recombinant baculovirus by site-specific transposition in E. colirather than homologous recombination in insect cells (Figure 1). This system features:
   Time-saving expression bacmid. With Bac-to-Bac®, the expression cassette of the pFastBac™ vector recombines with the parent bacmid in DH10Bac™ E. coli Competent Cells (not included with this vector kit) to form an expression bacmid. The bacmid is then transfected into insect cells for production of recombinant baculovirus particles.
   Easy colony screening. The parent bacmid in DH10Bac™ E. colicontains a segment of the lacZα gene. The lacZα gene is disrupted upon transposition of the expression cassette into the bacmid allowing for blue/white selection of recombinants. This makes identification of recombinant colonies easy.

 

pFastBacDual载体序列:

ORIGIN

        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga

       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc

      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct

      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg

      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga

      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt

      361 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg

      421 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc

      481 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat

      541 attaacgttt acaatttcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg

      601 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat

      661 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat

      721 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt

      781 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag

      841 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa

      901 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg

      961 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct

     1021 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac

     1081 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca

     1141 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat

     1201 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact

     1261 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc

     1321 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga

     1381 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg

     1441 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg

     1501 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca

     1561 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta

     1621 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca

     1681 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg

     1741 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga

     1801 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa

     1861 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc

     1921 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg

     1981 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac

     2041 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct

     2101 acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc

     2161 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg

     2221 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg

     2281 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct

     2341 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga

     2401 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg

     2461 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca

     2521 tctgtgcggt atttcacacc gcagaccagc cgcgtaacct ggcaaaatcg gttacggttg

     2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa

     2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga

     2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact

     2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg

     2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc

     2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac

     2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc

     3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag

     3061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat

     3121 ctcactacgc ggctgctcaa acctgggcag aacgtaagcc gcgagagcgc caacaaccgc

     3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta

     3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag

     3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat

     3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt

     3421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg

     3481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga

     3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg

     3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc

     3661 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg

     3721 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg

     3781 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg ccctggcttc

     3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag

     3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct

     3961 atagttctag tggttggcct acgtacccgt agtggctatg gcagggcttg ccgccccgac

     4021 gttggctgcg agccctgggc cttcacccga acttgggggt tggggtgggg aaaaggaaga

     4081 aacgcgggcg tattggtccc aatggggtct cggtggggta tcgacagagt gccagccctg

     4141 ggaccgaacc ccgcgtttat gaacaaacga cccaacaccc gtgcgtttta ttctgtcttt

     4201 ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc

     4261 tcccccatct cccggtaccg catgctatgc atcagctgct agcaccatgg ctcgagatcc

     4321 cgggtgatca agtcttcgtc gagtgattgt aaataaaatg taatttacag tatagtattt

     4381 taattaatat acaaatgatt tgataataat tcttatttaa ctataatata ttgtgttggg

     4441 ttgaattaaa ggtccgtata ctccggaata ttaatagatc atggagataa ttaaaatgat

     4501 aaccatctcg caaataaata agtattttac tgttttcgta acagttttgt aataaaaaaa

     4561 cctataaata ttccggatta ttcataccgt cccaccatcg ggcgcggatc ccggtccgaa

     4621 gcgcgcggaa ttcaaaggcc tacgtcgacg agctcactag tcgcggccgc tttcgaatct

     4681 agagcctgca gtctcgacaa gcttgtcgag aagtactaga ggatcataat cagccatacc

     4741 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa

     4801 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa

     4861 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt

     4921 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tctgatcact gcttgagcct

     4981 aggagatccg aaccagataa gtgaaatcta gttccaaact attttgtcat ttttaatttt

     5041 cgtattagct tacgacgcta cacccagttc ccatctattt tgtcactctt ccctaaataa

     5101 tccttaaaaa ctccatttcc acccctccca gttcccaact attttgtccg cccacagcgg

     5161 ggcatttttc ttcctgttat gtttttaatc aaacatcctg ccaactccat gtgacaaacc

     5221 gtcatcttcg gctacttt

//

 

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