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pComb3XTT

Details and Advantages
Applications: phage display system
概述 >

pComb3X is the newest of the pComb vectors. Improvements over pComb3 include increased stability and introduction of an asymmetric SfiI cassette for directional cloning of full Fab, scFv, peptide and other protein for phage display. 6xHis and HA tags allow for purification and detection. An amber stop codon was introduced to turn-off expression of the pIII fusion protein by switching to a non-supressor strain of E. coli allowing production of soluble protein without subcloning. Alternatively, the gene for phage protein pIII can be removed by SpeI/NheI digest. The “TT” refers to the human Fab to tetanus toxin which can be used as a template to amplify the kappa light chain constant region and Fd for construction of chimeric antibody libraries .


Summary

Bacterial Resistance(s): Ampicillin

  • Growth Temperature: 37°C

  • Growth Strain(s): TG1

  • Copy number: High Copy

  • Insert Size (bp): 1600

  • Promoter:lacZ

Tags / Fusion Proteins



     6x His (C terminal on backbone)

     HA tag (C terminal on backbone)

     geneIII (C terminal on backbone)

  • 5′ sequencing primer

    5'-AAG ACA GCT ATC GCG ATT GCA G-3'

  • 3′ sequencing primer

    5'-GCC CCC TTA TTA GCG TTT GCC ATC-3'

pcomb3XTT Map.png




















Vector sequence

        1 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg

       61 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta

      121 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg

      181 aattgtgagc ggataacaat tgaattcagg aggaatttaa aatgaaaaag acagctatcg

      241 cgattgcagt ggcactggct ggtttcgcta ccgtggccca ggcggccgag ctcacgcagt

      301 ctccaggcac cctgtctttg tctccagggg aaagagccac cctctcctgc agggccagtc

      361 acagtgttag cagggcctac ttagcctggt accagcagaa acctggccag gctcccaggc

      421 tcctcatcta tggtacatcc agcagggcca ctggcatccc agacaggttc agtggcagtg

      481 ggtctgggac agacttcact ctcaccatca gcagactgga gcctgaagat tttgcagtgt

      541 actactgtca gcagtatggt ggctcaccgt ggttcggcca agggaccaag gtggaactca

      601 aacgaactgt ggctgcacca tctgtcttca tcttcccgcc atctgatgag cagttgaaat

      661 ctggaactgc ctctgttgtg tgcctgctga ataacttcta tcccagagag gccaaagtac

      721 agtggaaggt ggataacgcc ctccaatcgg gtaactccca ggagagtgtc acagagcagg

      781 acagcaagga cagcacctac agcctcagca gcaccctgac gctgagcaaa gcagactacg

      841 agaaacacaa agtctacgcc tgcgaagtca cccatcaggg cctgagcttg cccgtcacaa

      901 agagcttcaa caggggagag tgttagttct agataattaa ttaggaggaa tttaaaatga

      961 aatacctatt gcctacggca gccgctggat tgttattact cgctgcccaa ccagccatgg

     1021 ccgaggtgca gctgctcgag cagtctgggg ctgaggtgaa gaagcctggg tcctcggtga

     1081 aggtctcctg cagggcttct ggaggcacct tcaacaatta tgccatcagc tgggtgcgac

     1141 aggcccctgg acaagggctt gagtggatgg gagggatctt ccctttccgt aatacagcaa

     1201 agtacgcaca acacttccag ggcagagtca ccattaccgc ggacgaatcc acgggcacag

     1261 cctacatgga gctgagcagc ctgagatctg aggacacggc catatattat tgtgcgagag

     1321 gggatacgat ttttggagtg accatgggat actacgctat ggacgtctgg ggccaaggga

     1381 ccacggtcac cgtctccgca gcctccacca agggcccatc ggtcttcccc ctggcaccct

     1441 cctccaagag cacctctggg ggcacagcgg ccctgggctg cctggtcaag gactacttcc

     1501 ccgaaccggt gacggtgtcg tggaactcag gcgccctgac cagcggcgtg cacaccttcc

     1561 cggctgtcct acagtcctca ggactctact ccctcagcag cgtggtgacc gtgccctcca

     1621 gcagcttggg cacccagacc tacatctgca acgtgaatca caagcccagc aacaccaagg

     1681 tggacaagaa agttgagccc aaatcttgtg acaaaactag tggccaggcc ggccagcacc

     1741 atcaccatca ccatggcgca tacccgtacg acgttccgga ctacgcttct taggagggtg

     1801 gtggctctga gggtggcggt tctgagggtg gcggctctga gggaggcggt tccggtggtg

     1861 gctctggttc cggtgatttt gattatgaaa agatggcaaa cgctaataag ggggctatga

     1921 ccgaaaatgc cgatgaaaac gtgctacagt ctgacgctaa aggcaaactt gattctgtcg

     1981 ctactgatta cggtgctgct atcgatggtt tcattggtga cgtttccggc cttgctaatg

     2041 gtaatggtgc tactggtgat tttgctggct ctaattccca aatggctcaa gtcggtgacg

     2101 gtgataattc acctttaatg aataatttcc gtcaatattt accttccctc cctcaatcgg

     2161 ttgaatgtcg cccttttgtc tttggcgctg gtaaaccata tgaattttct attgattgtg

     2221 acaaaataaa cttattccgt ggtgtctttg cgtttctttt atatgttgcc acctttatgt

     2281 atgtattttc tacgtttgct aacatactgc gtaataagga gtcttaagct agctaattaa

     2341 tttaagcggc cgcagatctg ctctctgagg aggatctggg aaattgtaag cgttaatatt

     2401 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa

     2461 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca

     2521 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc

     2581 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg

     2641 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg

     2701 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg

     2761 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg

     2821 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt

     2881 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg

     2941 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt

     3001 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta

     3061 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc

     3121 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa

     3181 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc

     3241 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt

     3301 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact

     3361 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac

     3421 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata

     3481 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta

     3541 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg

     3601 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat

     3661 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt

     3721 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga

     3781 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa

     3841 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag

     3901 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac

     3961 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc

     4021 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat

     4081 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat

     4141 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct

     4201 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt

     4261 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg

     4321 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta

     4381 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg

     4441 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg

     4501 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc

     4561 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg

     4621 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat

     4681 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc

     4741 agcgagtcag tgagcgagga agcgga

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