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基本信息

質粒圖譜和多克隆位點信息

注意：

客戶在我公司選擇構建載體服務項目時，請自行提供母源載體。

如要使用本公司保存的母源載體時，本公司不能保證所有母源載體完全符合客戶預期，也不承擔由此產生的任何責任。

載體描述

甘油醛-3-磷酸脫氫酶（GAPDH）在許多生物細胞中包括畢赤酵母中，穩定高水平表達。最近，GAPDH蛋白編碼基因啟動子（GAP）被深入研究，并在畢赤酵母中能夠高水表達重組蛋白，這取決于使用的碳源（Waterham等人， 1997）。GAP啟動子（PGAP）的重組蛋白表達水平略高于AOX1啟動子。

pGAPZ A，B，和C載體（2.9 KB）和pGAPZα A，B，和C（3.1 KB）載體使用GAP啟動子在畢赤酵母中穩定表達重組蛋白。重組蛋白表達為帶有C-末端myc抗原表位和C端His標簽的融合蛋白。此外，pGAPZα表達的融合蛋白在N-端帶有釀酒酵母（Saccharomyces cerevisiae）的α-因子分泌信號肽。這兩個系列的載體都提供三個不同讀碼框的載體，以方便克隆的帶有C-端標簽和/或N-末端分泌信號肽的載體中。

這些載體是基于顯性選擇標記，Zeocin（博萊霉素）的篩選標記可以同時使用于畢赤酵母和大腸桿菌。

載體序列

LOCUS       Exported                2884 bp ds-DNA     circular SYN 01-APR-2019

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    pGAPZA

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 2884)

  AUTHORS   BT-Lab

  TITLE     Direct Submission

  JOURNAL   Exported Monday, Apr 1, 2019 from SnapGene Viewer 4.3.6

FEATURES             Location/Qualifiers

     source          1..2884

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        7..483

                     /label=GAP promoter

                     /note="promoter for Pichia pastoris

                     glyceraldehyde-3-phosphate dehydrogenase"

     primer_bind     455..476

                     /label=pGAP-F

     misc_feature    484..563

                     /label=MCS

     CDS             564..593

                     /codon_start=1

                     /product="Myc (human c-Myc proto-oncogene) epitope tag"

                     /label=Myc

                     /translation="EQKLISEEDL"

     CDS             609..626

                     /codon_start=1

                     /product="6xHis affinity tag"

                     /label=6xHis

                     /translation="HHHHHH"

     terminator      705..951

                     /gene="Pichia pastoris AOX1"

                     /label=AOX1 terminator

                     /note="transcription terminator for AOX1"

     primer_bind     complement(711..731)

                     /label=3?AOX1

     promoter        966..1377

                     /gene="S. cerevisiae TEF1"

                     /label=TEF1 promoter

                     /note="promoter for EF-1-alpha"

     promoter        1384..1431

                     /label=EM7 promoter

                     /note="synthetic bacterial promoter "

     CDS             1450..1824

                     /codon_start=1

                     /gene="Sh ble from Streptoalloteichus hindustanus"

                     /product="antibiotic-binding protein"

                     /label=BleoR

                     /note="confers resistance to bleomycin, phleomycin, and

                     Zeocin(TM)"

                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD

                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR

                     EFALRDPAGNCVHFVAEEQD"

     terminator      1890..2137

                     /gene="S. cerevisiae CYC1"

                     /label=CYC1 terminator

                     /note="transcription terminator for CYC1"

     rep_origin      complement(2212..2800)

                     /direction=LEFT

                     /label=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 ccagcttggg cccgaacaaa aactcatctc agaagaggat ctgaatagcg

      601 ccgtcgacca tcatcatcat catcattgag ttttagcctt agacatgact gttcctcagt

      661 tcaagttggg cacttacgag aagaccggtc ttgctagatt ctaatcaaga ggatgtcaga

      721 atgccatttg cctgagagat gcaggcttca tttttgatac ttttttattt gtaacctata

      781 tagtatagga ttttttttgt cattttgttt cttctcgtac gagcttgctc ctgatcagcc

      841 tatctcgcag ctgatgaata tcttgtggta ggggtttggg aaaatcattc gagtttgatg

      901 tttttcttgg tatttcccac tcctcttcag agtacagaag attaagtgag accttcgttt

      961 gtgcggatcc cccacacacc atagcttcaa aatgtttcta ctcctttttt actcttccag

     1021 attttctcgg actccgcgca tcgccgtacc acttcaaaac acccaagcac agcatactaa

     1081 attttccctc tttcttcctc tagggtgtcg ttaattaccc gtactaaagg tttggaaaag

     1141 aaaaaagaga ccgcctcgtt tctttttctt cgtcgaaaaa ggcaataaaa atttttatca

     1201 cgtttctttt tcttgaaatt ttttttttta gtttttttct ctttcagtga cctccattga

     1261 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta

     1321 ttacaacttt ttttacttct tgttcattag aaagaaagca tagcaatcta atctaagggc

     1381 ggtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg

     1441 aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc

     1501 ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac

     1561 ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg

     1621 gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag

     1681 tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc

     1741 ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac

     1801 ttcgtggccg aggagcagga ctgacacgtc cgacggcggc ccacgggtcc caggcctcgg

     1861 agatccgtcc cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc

     1921 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta

     1981 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat

     2041 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct

     2101 tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag ctggagacca acatgtgagc

     2161 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag

     2221 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc

     2281 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt

     2341 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct

     2401 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg

     2461 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct

     2521 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat

     2581 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg

     2641 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa

     2701 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt

     2761 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc

     2821 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgcatga

     2881 gatc

//