P0302 pAN7-1 20μl

 真菌载体

P0302 pAN7-1 20μl "基本信息
启动子: gpdA
复制子: pUC ori
终止子: TRPC
质粒分类: 其他宿主载体；丝状真菌表达质粒
质粒大小: 6756bp
原核抗性: 氨苄青霉素Ampicillin
筛选标记: 潮霉素Hygromycin
克隆菌株: DH5α
培养条件: 37℃，有氧，LB
表达宿主: 丝状真菌
诱导方式: 无需诱导
5'测序引物: 根据全序列设计
3'测序引物: 根据全序列设计
备注: NCBI号：Z32698.1
质粒简介
参考文献
1、Effect of de-phosphorylation of linearized pAN7-1 and of addition of restriction enzyme on plasmid integration in Penicillium paxilli.
2、Transformation of sapstaining fungi with hygromycin B resistance plasmids pAN7-1 and pCB1004.
3、Padro de integrao de pAN7-1 em mutantes de Magnaporthe grisea com patogenicidade alterada em arroz.
4、Integração de pAN7-1 no Genoma de Magnaporthe grisea Mediada por Enzima de Restrição.
5、Primary structure of the trpC gene from Aspergillus nidulans.
6、Transformation of Aspergillus based on the hygromycin B resistance marker from Escherichia coli.
7、Functional elements in the promoter region of the Aspergillus nidulans gpdA gene encoding glyceraldehyde-3-phosphate dehydrogenase.

质粒图谱

质粒序列
LOCUS       Z32698                  6756 bp    DNA     linear   SYN 14-NOV-2006
DEFINITION  Expression vector (PAN7-1) DNA, 6756bp.
ACCESSION   Z32698
VERSION     Z32698.1  GI:475166
KEYWORDS    hygromycin b phosphotransferase.
SOURCE      synthetic construct
  ORGANISM  synthetic construct
            other sequences; artificial sequences.
REFERENCE   1
  AUTHORS   Mullaney,E.J., Hamer,J.E., Roberti,K.A., Yelton,M.M. and
            Timberlake,W.E.
  TITLE     Primary structure of the trpC gene from Aspergillus nidulans
  JOURNAL   Mol. Gen. Genet. 199 (1), 37-45 (1985)
   PUBMED   3158796
  REMARK    sites .
REFERENCE   2
  AUTHORS   Punt,P.J., Oliver,R.P., Dingemanse,M.A., Pouwels,P.H. and van den
            Hondel,C.A.
  TITLE     Transformation of Aspergillus based on the hygromycin B resistance
            marker from Escherichia coli
  JOURNAL   Gene 56 (1), 117-124 (1987)
   PUBMED   2824287
  REMARK    (sites)
REFERENCE   3  (bases 999 to 2572)
  AUTHORS   Punt,P.J., Dingemanse,M.A., Kuyvenhoven,A., Soede,R.D.,
            Pouwels,P.H. and van den Hondel,C.A.
  TITLE     Functional elements in the promoter region of the Aspergillus
            nidulans gpdA gene encoding glyceraldehyde-3-phosphate
            dehydrogenase
  JOURNAL   Gene 93 (1), 101-109 (1990)
   PUBMED   2121607
REFERENCE   4  (bases 1 to 6756)
  AUTHORS   Punt,P.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-APR-1994) Punt P., TNO Nutrition and Food Research
            Institute, Lange Kleiweg 139, Rijkswik, The Netherlands
FEATURES             Location/Qualifiers
     source          1..6756
                     /organism=""synthetic construct""
                     /mol_type=""other DNA""
                     /db_xref=""taxon:32630""
     misc_feature    1..6756
                     /product=""fungal transformation vector pAN7-1""
                     /standard_name=""HPH""
                     /experiment=""experimental evidence, no additional details
                     recorded""
                     /citation=[2]
                     /phenotype=""gene product confers resistance to hygromycin
                     B""
     regulatory      1..2129
                     /regulatory_class=""promoter""
                     /experiment=""experimental evidence, no additional details
                     recorded""
                     /citation=[2]
                     /citation=[3]
                     /function=""promoter region of the A.nidulans gpdA gene""
     gene            2130
                     /gene=""gpdA""
     misc_feature    2130
                     /gene=""gpdA""
                     /experiment=""experimental evidence, no additional details
                     recorded""
                     /citation=[3]
                     /function=""major transcription start site""
     intron          2178..2293
                     /experiment=""experimental evidence, no additional details
                     recorded""
                     /citation=[3]
                     /number=1
     gene            2302..3321
                     /gene=""HPH""
     CDS             2302..3321
                     /gene=""HPH""
                     /experiment=""experimental evidence, no additional details
                     recorded""
                     /note=""gene-product confers resistance to hygromycin B""
                     /citation=[2]
                     /citation=[3]
                     /codon_start=1
                     /transl_table=11
                     /product=""hygromycin B phosphotransferase""
                     /protein_id=""CAA83647.1""
                     /db_xref=""GI:475167""
                     /translation=""MPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEA
                     MFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSL
                     VDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTR
                     PRAKE""
     regulatory      3339..4108
                     /regulatory_class=""terminator""
                     /standard_name=""terminator region from A.nidulans trpC
                     gene (containing CDS)""
                     /experiment=""experimental evidence, no additional details
                     recorded""
                     /note=""this region contains the 3'end of the trpC
                     transcript including 50 codons. two major transcription
                     ends have been identified""
                     /citation=[1]
                     /citation=[2]
     polyA_site      3590..3600
     misc_difference 3600..3610
                     /note=""introduction of one base at this position generates
                     an BLGI site with was shown to be present by restriction
                     digestion""
                     /citation=[1]
                     /replace=""gccttncaggc""
     misc_difference 3809..3817
                     /note=""introduction of one base at this position generates
                     an ALWNI site which was shown to be present by restriction
                     digestion""
                     /citation=[1]
                     /replace=""cagcncctg""
     polyA_site      3863..3873
     misc_difference 3897..3907
                     /note=""introduction (actual sequence could be deletion as
                     well) of a base at this position results in the
                     elimination of an XMNI site which was shown to be absent
                     by restriction digestion""
                     /citation=[1]
                     /replace=""gaaatcanttc""
     variation       3934..3941
                     /note=""A NCOI site present at this position was removed by
                     digestion, treatment with klenow polymerase and
                     religation""
                     /citation=[2]
                     /replace=""catgcatg""
     misc_feature    4105..6756
                     /product=""PUC18""
                     /note=""vector sequences derived from PUC18 from SALI TO
                     ECORI""
                     /citation=[2]
ORIGIN     
        1 gaattccctt gtatctctac acacaggctc aaatcaataa gaagaacggt tcgtcttttt
       61 cgtttatatc ttgcatcgtc ccaaagctat tggcgggata ttctgtttgc agttggctga
      121 cttgaagtaa tctctgcaga tctttcgaca ctgaaatacg tcgagcctgc tccgcttgga
      181 agcggcgagg agcctcgtcc tgtcacaact accaacatgg agtacgataa gggccagttc
      241 cgccagctca ttaagagcca gttcatgggc gttggcatga tggccgtcat gcatctgtac
      301 ttcaagtaca ccaacgctct tctgatccag tcgatcatcc gctgaaggcg ctttcgaatc
      361 tggttaagat ccacgtcttc gggaagccag cgactggtga cctccagcgt ccctttaagg
      421 ctgccaacag ctttctcagc cagggccagc ccaagaccga caaggcctcc ctccagaacg
      481 ccgagaagaa ctggaggggt ggtgtcaagg aggagtaagc tccttattga agtcggagga
      541 cggagcggtg tcaagaggat attcttcgac tctgtattat agataagatg atgaggaatt
      601 ggaggtagca tagcttcatt tggatttgct ttccaggctg agactctagc ttggagcata
      661 gagggtcctt tggctttcaa tattctcaag tatctcgagt ttgaacttat tccctgtgaa
      721 ccttttattc accaatgagc attggaatga acatgaatct gaggactgca atcgccatga
      781 ggttttcgaa atacatccgg atgtcgaagg cttggggcac ctgcgttggt tgaatttaga
      841 acgtggcact attgatcatc cgatagctct gcaaagggcg ttgcacaatg caagtcaaac
      901 gttgctagca gttccaggtg gaatgttatg atgagcattg tattaaatca ggagatatag
      961 catgatctct agttagctca ccacaaaagt cagacggcgt aaccaaaagt cacacaacac
     1021 aagctgtaag gatttcggca cggctacgga agacggagaa gccaccttca gtggactcga
     1081 gtaccattta attctatttg tgtttgatcg agacctaata cagcccctac aacgaccatc
     1141 aaagtcgtat agctaccagt gaggaagtgg actcaaatcg acttcagcaa catctcctgg
     1201 ataaacttta agcctaaact atacagaata agataggtgg agagcttata ccgagctccc
     1261 aaatctgtcc agatcatggt tgaccggtgc ctggatcttc ctatagaatc atccttattc
     1321 gttgacctag ctgattctgg agtgacccag agggtcatga cttgagccta aaatccgccg
     1381 cctccaccat ttgtagaaaa atgtgacgaa ctcgtgagct ctgtacagtg accggtgact
     1441 ctttctggca tgcggagaga cggacggacg cagagagaag ggctgagtaa taagccactg
     1501 gccagacagc tctggcggct ctgaggtgca gtggatgatt attaatccgg gaccggccgc
     1561 ccctccgccc cgaagtggaa aggctggtgt gcccctcgtt gaccaagaat ctattgcatc
     1621 atcggagaat atggagcttc atcgaatcac cggcagtaag cgaaggagaa tgtgaagcca
     1681 ggggtgtata gccgtcggcg aaatagcatg ccattaacct aggtacagaa gtccaattgc
     1741 ttccgatctg gtaaaagatt cacgagatag taccttctcc gaagtaggta gagcgagtac
     1801 ccggcgcgta agctccctaa ttggcccatc cggcatctgt agggcgtcca aatatcgtgc
     1861 ctctcctgct ttgcccggtg tatgaaaccg gaaaggccgc tcaggagctg gccagcggcg
     1921 cagaccggga acacaagctg gcagtcgacc catccggtgc tctgcactcg acctgctgag
     1981 gtccctcagt ccctggtagg cagctttgcc ccgtctgtcc gcccggtgtg tcggcggggt
     2041 tgacaaggtc gttgcgtcag tccaacattt gttgccatat tttcctgctc tccccaccag
     2101 ctgctctttt cttttctctt tcttttccca tcttcagtat attcatcttc ccatccaaga

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