• 商品货号：FH1679

• 规格：10μl质粒
• 质粒图谱1： png/1-1G12611462X44.png

质粒序列： 点击查看序列

LOCUS       Exported                9294 bp ds-DNA     circular SYN 26-NOV-2017

DEFINITION  synthetic circular DNA

FEATURES             Location/Qualifiers

     source          1..9294

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        2..937

                     /gene="Pichia pastoris AOX1"

                     /label=AOX1 promoter

                     /note="inducible promoter, regulated by methanol"

     CDS             949..1215

                     /codon_start=1

                     /gene="MF-alpha-1"

                     /product="N-terminal secretion signal from S. cerevisiae 

                     alpha-factor"

                     /label=-alpha-factor secretion signal

                     /note="Cleavage by the Kex2 protease occurs after the 

                     dibasic KR sequence. The EA dipeptides are then removed by 

                     dipeptidyl aminopeptidase A."

                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE

                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"

     misc_feature    1216..1241

                     /label=MCS

     misc_feature    1246..1263

                     /label=6×HIS

     terminator      1339..1585

                     /gene="Pichia pastoris AOX1"

                     /label=AOX1 terminator

                     /note="transcription terminator for AOX1"

     CDS             complement(1998..4532)

                     /codon_start=1

                     /gene="Pichia pastoris HIS4"

                     /product="multifunctional enzyme, required for histidine 

                     biosynthesis"

                     /label=PpHIS4

                     /note="auxotrophic marker for Pichia pastoris"

                     /translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQSTE

                     LQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYRTL

                     VDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGIIPII

                     DARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSSKKSI

                     PEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGFCHLER

                     TSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAEAKSKED

                     IAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQPKEESKP

                     KEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQNGDKALLE

                     LTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLTETLQVETCP

                     GVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASPPKKDGTLTPE

                     VIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTAAKMMVQNDTSA

                     LCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAVDMTDKELARIED

                     AVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLILQIENASSYVDQV

                     QHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQKFITSQDVTPEGLK

                     HIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI"

     CDS             complement(4946..5761)

                     /codon_start=1

                     /gene="aph(3')-Ia"

                     /product="aminoglycoside phosphotransferase"

                     /label=KanR

                     /note="confers resistance to kanamycin in bacteria or G418 

                     (Geneticin(R)) in eukaryotes"

                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG

                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK

                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA

                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI

                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"

     misc_feature    6140..6896

                     /label=AOX1 3' fragment

                     /note="region downstream of Pichia pastoris AOX1 gene"

     misc_feature    7039..7179

                     /label=bom

                     /note="basis of mobility region from pBR322"

     rep_origin      complement(7365..7953)

                     /direction=LEFT

                     /label=ori

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 

                     replication"

     CDS             complement(8124..8984)

                     /codon_start=1

                     /gene="bla"

                     /product="beta-lactamase"

                     /label=AmpR

                     /note="confers resistance to ampicillin, carbenicillin, and

                     related antibiotics"

                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS

                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

                     LIKHW"

     promoter        complement(8985..9089)

                     /gene="bla"

                     /label=AmpR promoter

ORIGIN

        1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag

       61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt

      121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc

      181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta

      241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta

      301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg

      361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct

      421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg

      481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcgcca taccgtttgt

      541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct

      601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct

      661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact

      721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat

      781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt

      841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga

      901 caacttgaga agatcaaaaa acaactaatt attcgaagga tccaaacgat gagatttcct

      961 tcaattttta ctgcagtttt attcgcagca tcctccgcat tagctgctcc agtcaacact

     1021 acaacagaag atgaaacggc acaaattccg gctgaagctg tcatcggtta ctcagattta

     1081 gaaggggatt tcgatgttgc tgttttgcca ttttccaaca gcacaaataa cgggttattg

     1141 tttataaata ctactattgc cagcattgct gctaaagaag aaggggtatc tctcgagaaa

     1201 agagaggctg aagcttacgt agaattccct agggcggccg cgaatcatca tcaccatcac

     1261 cattaattcg ccttagacat gactgttcct cagttcaagt tgggcactta cgagaagacc

     1321 ggtcttgcta gattctaatc aagaggatgt cagaatgcca tttgcctgag agatgcaggc

     1381 ttcatttttg atactttttt atttgtaacc tatatagtat aggatttttt ttgtcatttt

     1441 gtttcttctc gtacgagctt gctcctgatc agcctatctc gcagctgatg aatatcttgt

     1501 ggtaggggtt tgggaaaatc attcgagttt gatgtttttc ttggtatttc ccactcctct

     1561 tcagagtaca gaagattaag tgagaagttc gtttgtgcaa gcttatcgat aagctttaat

     1621 gcggtagttt atcacagtta aattgctaac gcagtcaggc accgtgtatg aaatctaaca

     1681 atgcgctcat cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata ggcttggtta

     1741 tgccggtact gccgggcctc ttgcgggata tcgtccattc cgacagcatc gccagtcact

     1801 atggcgtgct gctagcgcta tatgcgttga tgcaatttct atgcgcaccc gttctcggag

     1861 cactgtccga ccgctttggc cgccgcccag tcctgctcgc ttcgctactt ggagccacta

     1921 tcgactacgc gatcatggcg accacacccg tcctgtggat ctatcgaatc taaatgtaag

     1981 ttaaaatctc taaataatta aataagtccc agtttctcca tacgaacctt aacagcattg

     2041 cggtgagcat ctagaccttc aacagcagcc agatccatca ctgcttggcc aatatgtttc

     2101 agtccctcag gagttacgtc ttgtgaagtg atgaacttct ggaaggttgc agtgttaact

     2161 ccgctgtatt gacgggcata tccgtacgtt ggcaaagtgt ggttggtacc ggaggagtaa

     2221 tctccacaac tctctggaga gtaggcacca acaaacacag atccagcgtg ttgtacttga

     2281 tcaacataag aagaagcatt ctcgatttgc aggatcaagt gttcaggagc gtactgattg

     2341 gacatttcca aagcctgctc gtaggttgca accgataggg ttgtagagtg tgcaatacac

     2401 ttgcgtacaa tttcaaccct tggcaactgc acagcttggt tgtgaacagc atcttcaatt

     2461 ctggcaagct ccttgtctgt catatcgaca gccaacagaa tcacctggga atcaatacca

     2521 tgttcagctt gagacagaag gtctgaggca acgaaatctg gatcagcgta tttatcagca

     2581 ataactagaa cttcagaagg cccagcaggc atgtcaatac tacacagggc tgatgtgtca

     2641 ttttgaacca tcatcttggc agcagtaacg aactggtttc ctggaccaaa tattttgtca

     2701 cacttaggaa cagtttctgt tccgtaagcc atagcagcta ctgcctgggc gcctcctgct

     2761 agcacgatac acttagcacc aaccttgtgg gcaacgtaga tgacttctgg ggtaagggta

     2821 ccatccttct taggtggaga tgcaaaaaca atttctttgc aaccagcaac tttggcagga

     2881 acacccagca tcagggaagt ggaaggcaga attgcggttc caccaggaat atagaggcca

     2941 actttctcaa taggtcttgc aaaacgagag cagactacac cagggcaagt ctcaacttgc

     3001 aacgtctccg ttagttgagc ttcatggaat ttcctgacgt tatctataga gagatcaatg

     3061 gctctcttaa cgttatctgg caattgcata agttcctctg ggaaaggagc ttctaacaca

     3121 ggtgtcttca aagcgactcc atcaaacttg gcagttagtt ctaaaagggc tttgtcacca

     3181 ttttgacgaa cattgtcgac aattggtttg actaattcca taatctgttc cgttttctgg

     3241 ataggacgac gaagggcatc ttcaatttct tgtgaggagg ccttagaaac gtcaattttg

     3301 cacaattcaa tacgaccttc agaagggact tctttaggtt tggattcttc tttaggttgt

     3361 tccttggtgt atcctggctt ggcatctcct ttccttctag tgacctttag ggacttcata

     3421 tccaggtttc tctccacctc gtccaacgtc acaccgtact tggcacatct aactaatgca

     3481 aaataaaata agtcagcaca ttcccaggct atatcttcct tggatttagc ttctgcaagt

     3541 tcatcagctt cctccctaat tttagcgttc aacaaaactt