pGAPZB Plasmid

$136.00

  • Model: PVT4010
  • 50 Units in Stock
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pGAPZB

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pGAPZB,Plasmid pGAPZB,pGAPZB vector

 

pGAPZB Information

Bacterial Resistance:Zeo
Growth Strain:
Expression: Yeast
Use: Yeast expression

 

pGAPZB  Desciption

Glyceraldehyde -3- phosphate dehydrogenase (GAPDH) enzyme is stable and highly expressed in many biological cells including Pichia pastoris. Recently, the GAPDH protein coding gene promoter (GAP) has been studied in depth and can be expressed in high water in Pichia pastoris, depending on the use of carbon sources (Waterham et al., 1997). The expression level of GAP promoter (PGAP) was slightly higher than that of AOX1 promoter.

PGAPZ A, B, and C vectors (2.9 KB) and pGAPZ alpha, B, and C (3.1 KB) vectors were used to stably express recombinant proteins in the Pichia pastoris using a GAP promoter. The recombinant protein was expressed as a fusion protein with C- terminal myc epitope and C terminal His tag. In addition, the fusion protein pGAPZ alpha expresses at the N- end with the alpha factor secreting signal peptide of Saccharomyces cerevisiae. These two series of vectors provide three different reading frame carriers to facilitate cloning of vectors carrying C- terminal tags and / or N- terminal secretory signal peptides. These vectors are based on dominant selection markers, and Zeocin (bleomycin) screening markers can be used in Pichia pastoris and Escherichia coli simultaneously.

 

pGAPZB Sequence

LOCUS       Exported                2882 bp ds-DNA     circular SYN 10-SEP-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2882)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Saturday, September 10, 2016 from SnapGene Viewer 3.1.4
FEATURES             Location/Qualifiers
     source          1..2882
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        7..483
                     /note="GAP promoter"
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             562..591
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /note="Myc"
                     /translation="EQKLISEEDL"
     CDS             607..624
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     terminator      703..949
                     /gene="Pichia pastoris AOX1"
                     /note="AOX1 terminator"
                     /note="transcription terminator for AOX1"
     promoter        964..1375
                     /gene="S. cerevisiae TEF1"
                     /note="TEF1 promoter"
                     /note="promoter for EF-1-alpha"
     promoter        1382..1429
                     /note="EM7 promoter"
                     /note="synthetic bacterial promoter "
     CDS             1448..1822
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /note="BleoR"
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      1888..2135
                     /gene="S. cerevisiae CYC1"
                     /note="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      complement(2210..2798)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agatcttttt tgtagaaatg tcttggtgtc ctcgtccaat caggtagcca tctctgaaat
       61 atctggctcc gttgcaactc cgaacgacct gctggcaacg taaaattctc cggggtaaaa
      121 cttaaatgtg gagtaatgga accagaaacg tctcttccct tctctctcct tccaccgccc
      181 gttaccgtcc ctaggaaatt ttactctgct ggagagcttc ttctacggcc cccttgcagc
      241 aatgctcttc ccagcattac gttgcgggta aaacggaggt cgtgtacccg acctagcagc
      301 ccagggatgg aaaagtcccg gccgtcgctg gcaataatag cgggcggacg catgtcatga
      361 gattattgga aaccaccaga atcgaatata aaaggcgaac acctttccca attttggttt
      421 ctcctgaccc aaagacttta aatttaattt atttgtccct atttcaatca attgaacaac
      481 tatttcgaaa cgaggaattc acgtggccca gccggccgtc tcggatcggt acctcgagcc
      541 gcggcggccg ccagctttct agaacaaaaa ctcatctcag aagaggatct gaatagcgcc
      601 gtcgaccatc atcatcatca tcattgagtt ttagccttag acatgactgt tcctcagttc
      661 aagttgggca cttacgagaa gaccggtctt gctagattct aatcaagagg atgtcagaat
      721 gccatttgcc tgagagatgc aggcttcatt tttgatactt ttttatttgt aacctatata
      781 gtataggatt ttttttgtca ttttgtttct tctcgtacga gcttgctcct gatcagccta
      841 tctcgcagct gatgaatatc ttgtggtagg ggtttgggaa aatcattcga gtttgatgtt
      901 tttcttggta tttcccactc ctcttcagag tacagaagat taagtgagac cttcgtttgt
      961 gcggatcccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
     1021 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
     1081 tttccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
     1141 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
     1201 tttctttttc ttgaaatttt tttttttagt ttttttctct ttcagtgacc tccattgata
     1261 tttaagttaa taaacggtct tcaatttctc aagtttcagt ttcatttttc ttgttctatt
     1321 acaacttttt ttacttcttg ttcattagaa agaaagcata gcaatctaat ctaagggcgg
     1381 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     1441 ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg
     1501 agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt
     1561 cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt
     1621 gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg
     1681 gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg
     1741 cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt
     1801 cgtggccgag gagcaggact gacacgtccg acggcggccc acgggtccca ggcctcggag
     1861 atccgtcccc cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac
     1921 gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     1981 tccctattta tttttttata gttatgttag tattaagaac gttatttata tttcaaattt
     2041 ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg
     2101 agaaggtttt gggacgctcg aaggctttaa tttgcaagct ggagaccaac atgtgagcaa
     2161 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2221 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2281 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2341 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2401 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2461 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2521 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2581 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2641 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2701 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
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