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pComb3XLambda

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. pComb3XLambda can be used as a template to amplify the lambda light chain constant region for construction of chimeric antibody libraries as described in: Barbas, C. F., III; Burton, D. R.; Scott, J.K., Silverman, G.J. Eds. (2001) Phage Display: A Laboratory Manual; Cold Spring Harbor Laboratory Press: Cold Spring Harbor, New York

Please note that there is a A246V mutation in heavy chain region; however, this vector is meant to be used as a PCR template for the light chain region from base# 613-1016.

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

  • HA tag (C terminal on backbone)

  • 6x His (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'

pComb3XLambda.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 ctcgtgctga

      301 ctaacgccct cagtgtctgg ggccccaggg cagagggtca ccatctcctg cactgggagc

      361 agctccaacc tcggggcagc ttatgatgta cactggtacc accaggttcc aggaacggcc

      421 cccaaactcc tcctctatga taacatcaag cggccctcag gggtccctga ccgattctct

      481 ggctccaagt ctggcacctc agcctccctg gccatcactg ggctccaggc tgaggacgag

      541 gctgattatt actgcaacag agtaacagct ggcctcacac gttcggcgga gggaccaagc

      601 tgaccgtcct aagtcagccc aaggctgccc cctcggtcac tctgttcccg ccctcctctg

      661 aggagcttca agccaacaag gccacactgg tgtgtctcat aagtgacttc tacccgggag

      721 ccgtgacagt ggcctggaag gcagatggca gccccgtcaa ggcgggagtg gagaccacca

      781 caccctccaa acaaagcaac aacaagtacg cggccagcag ctatctgagc ctgacgcctg

      841 agcagtggaa gtcccacaga agctacagct gccaggtcac gcatgaaggg agcaccgtgg

      901 agaagacagt ggcccctaca gaatgttcat aattctagat aattaattag gaggaattta

      961 aaatgaaata cctattgcct acggcagccg ctggattgtt attactcgct gcccaaccag

     1021 ccatggccga ggtgcagctg ctcgagcagt ctggggctga ggtgaagaag cctgggtcct

     1081 cggtgaaggt ctcctgcagg gcttctggag gcaccttcaa caattatgcc atcagctggg

     1141 tgcgacaggc ccctggacaa gggcttgagt ggatgggagg gatcttccct ttccgtaata

     1201 cagcaaagta cgcacaacac ttccagggca gagtcaccat taccgcggac gaatccacgg

     1261 gcacagccta catggagctg agcagcctga gatctgagga cacggccata tattattgtg

     1321 cgagagggga tacgattttt ggagtgacca tgggatacta cgctatggac gtctggggcc

     1381 aagggaccac ggtcaccgtc tccgcagcct ccaccaaggg cccatcggtc ttccccctgg

     1441 caccctcctc caagagcacc tctgggggca cagcggccct gggctgcctg gtcaaggact

     1501 acttccccga accggtgacg gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca

     1561 ccttcccggc tgtcctacag tcctcaggac tctactccct cagcagcgtg gtgaccgtgc

     1621 cctccagcag cttgggcacc cagacctaca tctgcaacgt gaatcacaag cccagcaaca

     1681 ccaaggtgga caagaaagtt gagcccaaat cttgtgacaa aactagtggc caggccggcc

     1741 agcaccatca ccatcaccat ggcgcatacc cgtacgacgt tccggactac gcttcttagg

     1801 agggtggtgg ctctgagggt ggcggttctg agggtggcgg ctctgaggga ggcggttccg

     1861 gtggtggctc tggttccggt gattttgatt atgaaaagat ggcaaacgct aataaggggg

     1921 ctatgaccga aaatgccgat gaaaacgtgc tacagtctga cgctaaaggc aaacttgatt

     1981 ctgtcgctac tgattacggt gctgctatcg atggtttcat tggtgacgtt tccggccttg

     2041 ctaatggtaa tggtgctact ggtgattttg ctggctctaa ttcccaaatg gctcaagtcg

     2101 gtgacggtga taattcacct ttaatgaata atttccgtca atatttacct tccctccctc

     2161 aatcggttga atgtcgccct tttgtctttg gcgctggtaa accatatgaa ttttctattg

     2221 attgtgacaa aataaactta ttccgtggtg tctttgcgtt tcttttatat gttgccacct

     2281 ttatgtatgt attttctacg tttgctaaca tactgcgtaa taaggagtct taagctagct

     2341 aattaattta agcggccgca gatctgctct ctgaggagga tctgggaaat tgtaagcgtt

     2401 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag

     2461 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt

     2521 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga

     2581 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg

     2641 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct

     2701 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc

     2761 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt

     2821 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct

     2881 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga

     2941 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc

     3001 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg

     3061 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc

     3121 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact

     3181 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc

     3241 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag

     3301 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat

     3361 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt

     3421 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa

     3481 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc

     3541 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg

     3601 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt

     3661 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca

     3721 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat

     3781 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca

     3841 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg

     3901 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg

     3961 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt

     4021 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg

     4081 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata

     4141 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca

     4201 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag

     4261 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc

     4321 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga

     4381 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg

     4441 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac

     4501 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg

     4561 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg

     4621 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct

     4681 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc

     4741 gagcgcagcg agtcagtgag cgaggaagcg ga

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