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ID   PAN24      preliminary; circular DNA; SYN; 5760 BP.
XX
AC   Z32750;
XX
DT   01-JUL-1994 (Rel. 9, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE   E. coli plasmid vector pAN2-4 or pAN52-5NOT - complete.
XX
KW   cloning vector.
XX
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
XX
RN   [1]
RP   999-2572
RC   M13mAN5-7B from M13mp11 & gpdA gene
RC   from pAN5-41B
RC   from pAN923-41B
RC   from pAN923-42B
RA   Punt P.J., Dingemanse M., Kuyvenhoven A., Soede R.D.M.,
RA   Pouwels P.H., Van Den Hondel C.A.M.J.J.;
RT   "Functional elements in the promoter region of the Aspergillus
RT   nidulans GPDA gene encoding glyceraldehyde-3-phosphate
RT   dehydrogenase";
RL   Gene 93:101-109(1990).
XX
RN   [2]
RP   1-5760
RC   pAN series from gpdA gene
RA   Punt P.J., Kuijvenhoven A., Broekhuijsen M.P.,
RA   Van Den Hondel C.A.M.J.J.;
RT   "Vectors for efficient production of homologous and heterologous
RT   proteins in filamentous fungi";
RL   Unpublished (1994).
XX
RN   [3]
RP   2305-3071
RC   from A.nidulans trpC gene
RA   Mullaney E.J., Hamer J.E., Roberti K.A., Yelton M.,
RA   Timberlake W.E.;
RT   "Primary structure of the trpC gene from Aspergillus nidulans";
RL   Mol. Gen. Genet. 199:37-45(1985).
XX
RN   [4]
RP   1-5760
RC   pAN2-4 or pAN52-5NOT
RA   Punt P.;
RT   ;
RL   Submitted (22-APR-1994) to the EMBL/GenBank/DDBJ databases by:
RL   Punt P., TNO Nutrition and Food Research Institute, Lange Kleiweg 139,
RL   Rijkswik, The Netherlands.
XX
RN   [5]
RC   pYS4 from A.nidulans trpC gene
RA   Yelton M., Hamer J.E., DeSouza E.R., Mullaney E.J., Timberlake W.E.;
RT   "Developmental regulation of the Aspergillus nidulans trpC gene";
RL   Proc. Natl. Acad. Sci. U.S.A. 80:7576-7580(1983).
XX
RN   [6]
RC   pHY201 from A.nidulans trpC gene
RA   Yelton M., Hamer J.E., Timberlake W.E.;
RT   "Transformation of Aspergillus nidulans by using a trpC plasmid";
RL   Proc. Natl. Acad. Sci. U.S.A. 81:1470-1474(1984).
XX
RN   [7]
RC   pAN923-41B, pAN923-42B from argB gene
RC   pAN5-41B from A.nidulans gpdA gene
RA   Van Gorcom R.F., Punt P.J., Pouwels P.H., Van Den Hondel C.A.;
RT   "A system for the analysis of expression signals in Aspergillus";
RL   Gene 48:211-217(1986).
XX
RN   [8]
RC   pAB94-11 from pyrG* gene
RA   Van Gorcom R.F., Van Den Hondel C.A.;
RT   "Expression analysis vectors for Aspergillus niger";
RL   Nucleic Acids Res. 18:9052-9052(1988).
XX
RN   [9]
RC   pAN52-1NotI from A.nidulans gpdA gene
RC   pAN52-6NotI from A.niger glaA gene
RA   Van Den Hondel C.A., Punt P., Van Gorcom R.F.;
RT   "Heterologous gene expression in filamentous fungi";
RL   More Gene Manipulations in Fungi 0:396-428(1991).
RL   Benneth J.W., Lasure L.L., eds., Academic Press, San Diego CA.
XX
CC   NCBI gi: 475897
CC   NM (pAN2-4)(pAN52-5NOT)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP ()
CC   HO (E.coli)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA (pAN52-5NOT)
CC   BR ()
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pUC18 EcoRI-HindIII 2635bp 451..2686..400
FT                   2. A.nidulans HindIII-EcoRI 3120bp, gpdA gene
FT                   -> pAN2-4 5760bp"
FT   promoter        1..2129
FT                   /note="PRO A. nidulans gpdA gene;
FT                   EXPERIMENTAL [1],[2]"
FT   misc_feature    2130..2130
FT                   /note="A. nidulans gpdA gene major
FT                   transcription start site; EXPERIMENTAL [1]"
FT   intron          2178..2293
FT                   /note="EXPERIMENTAL [1]"
FT   terminator      2392..3115
FT                   /note="TER A. nidulans trpC gene terminator region
FT                   (containing CDS); EXPERIMENTAL [2],[3];
FT                   contains the 3'end of the trpC
FT                   transcript including 50 codons; two major
FT                   transcription ends have been identified"
FT   misc_difference 2652..2662
FT                   /note="replace(2652..2662,gccttncaggc);
FT                   introduction of one base at this position generates
FT                   an BlgI site which was shown to be present by
FT                   restriction digestion [3]"
FT   misc_difference 2861..2869
FT                   /note="replace(2861..2869,cagcncctg);
FT                   introduction of one base at this position generates
FT                   an AlwNI site which was shown to be present by
FT                   restriction digestion [3]"
FT   misc_difference 2949..2959
FT                   /note="replace(2949..2959,gaaatcanttc);
FT                   introduction (actual sequence could be deletion as
FT                   well) of a base at this position results in the
FT                   elimination of an XmnI site which was shown to be
FT                   absent by restriction digestion [3]"
FT   mutation        2986..2993
FT                   /note="replace(2986..2993,catgcatg);
FT                   A NcoI site present at this position was removed by
FT                   digestion, treatment with klenow polymerase and
FT                   religation [2]"
FT   polyA_site      2642..2652
FT                   /note="partial"
FT   polyA_site      2915..2925
FT                   /note="partial"
FT   misc_feature    3115..5753
FT                   /note="from pUC18 HindIII-EcoRI [2]"
FT   misc_feature    1..5760
FT                   /note="fungal expression/secretion vector pAN52-5NOT;
FT                   EXPERIMENTAL [2]"
FT   sig_peptide     2302..2355
FT                   /note="synthetic signal sequence, very similar to
FT                   that of the signal sequence of the
FT                   A. niger glucoamylase gene (accession #X00712) [2]"
FT   misc_feature    2356..2375
FT                   /note="PRO_PEPTIDE sequence for putative propeptide;
FT                   contains dibasic processing site (LYS-ARG) [2]"
FT   CDS             2302..2391
FT                   /note="gpdA secretion signal sequence very similar to
FT                   that of the A.niger glucoamylase gene;
FT                   NCBI gi: 475898; EXPERIMENTAL [2]"
XX
SQ   Sequence 5760 BP; 1437 A; 1458 C; 1391 G; 1471 T; 3 other;
     caattccctt gtatctctac acacaggctc aaatcaataa gaagaacggt tcgtcttttt
     cgtttatatc ttgcatcgtc ccaaagctat tggcgggata ttctgtttgc agttggctga
     cttgaagtaa tctctgcaga tctttcgaca ctgaaatacg tcgagcctgc tccgcttgga
     agcggcgagg agcctcgtcc tgtcacaact accaacatgg agtacgataa gggccagttc
     cgccagctca ttaagagcca gttcatgggc gttggcatga tggccgtcat gcatctgtac
     ttcaagtaca ccaacgctct tctgatccag tcgatcatcc gctgaaggcg ctttcgaatc
     tggttaagat ccacgtcttc gggaagccag cgactggtga cctccagcgt ccctttaagg
     ctgccaacag ctttctcagc cagggccagc ccaagaccga caaggcctcc ctccagaacg
     ccgagaagaa ctggaggggt ggtgtcaagg aggagtaagc tccttattga agtcggagga
     cggagcggtg tcaagaggat attcttcgac tctgtattat agataagatg atgaggaatt
     ggaggtagca tagcttcatt tggatttgct ttccaggctg agactctagc ttggagcata
     gagggtcctt tggctttcaa tattctcaag tatctcgagt ttgaacttat tccctgtgaa
     ccttttattc accaatgagc attggaatga acatgaatct gaggactgca atcgccatga
     ggttttcgaa atacatccgg atgtcgaagg cttggggcac ctgcgttggt tgaatttaga
     acgtggcact attgatcatc cgatagctct gcaaagggcg ttgcacaatg caagtcaaac
     gttgctagca gttccaggtg gaatgttatg atgagcattg tattaaatca ggagatatag
     catgatctct agttagctca ccacaaaagt cagacggcgt aaccaaaagt cacacaacac
     aagctgtaag gatttcggca cggctacgga agacggagaa gccaccttca gtggactcga
     gtaccattta attctatttg tgtttgatcg agacctaata cagcccctac aacgaccatc
     aaagtcgtat agctaccagt gaggaagtgg actcaaatcg acttcagcaa catctcctgg
     ataaacttta agcctaaact atacagaata agataggtgg agagcttata ccgagctccc
     aaatctgtcc agatcatggt tgaccggtgc ctggatcttc ctatagaatc atccttattc
     gttgacctag ctgattctgg agtgacccag agggtcatga cttgagccta aaatccgccg
     cctccaccat ttgtagaaaa atgtgacgaa ctcgtgagct ctgtacagtg accggtgact
     ctttctggca tgcggagaga cggacggacg cagagagaag ggctgagtaa taagccactg
     gccagacagc tctggcggct ctgaggtgca gtggatgatt attaatccgg gaccggccgc
     ccctccgccc cgaagtggaa aggctggtgt gcccctcgtt gaccaagaat ctattgcatc
     atcggagaat atggagcttc atcgaatcac cggcagtaag cgaaggagaa tgtgaagcca
     ggggtgtata gccgtcggcg aaatagcatg ccattaacct aggtacagaa gtccaattgc
     ttccgatctg gtaaaagatt cacgagatag taccttctcc gaagtaggta gagcgagtac
     ccggcgcgta agctccctaa ttggcccatc cggcatctgt agggcgtcca aatatcgtgc
     ctctcctgct ttgcccggtg tatgaaaccg gaaaggccgc tcaggagctg gccagcggcg
     cagaccggga acacaagctg gcagtcgacc catccggtgc tctgcactcg acctgctgag
     gtccctcagt ccctggtagg cagctttgcc ccgtctgtcc gcccggtgtg tcggcggggt
     tgacaaggtc gttgcgtcag tccaacattt gttgccatat tttcctgctc tccccaccag
     ctgctctttt cttttctctt tcttttccca tcttcagtat attcatcttc ccatccaaga
     acctttattt cccctaagta agtactttgc tacatccata ctccatcctt cccatccctt
     attcctttga acctttcagt tcgagctttc ccacttcatc gcagcttgac taacagctac
     cccgcttgag cagacatcac catgggcttc cgatctctac tcgccctgag cggcctcgtc
     tgcaacgggt tggcaaatgt gatttccaag cgcgctggat ccgaagcttg agatccactt
     aacgttactg aaatcatcaa acagcttgac gaatctggat ataagatcgt tggtgtcgat
     gtcagctccg gagttgagac aaatggtgtt caggatctcg ataagatacg ttcatttgtc
     caagcagcaa agagtgcctt ctagtgattt aatagctcca tgtcaacaag aataaaacgc
     gttttcgggt ttacctcttc cagatacagc tcatctgcaa tgcattaatg cattgactgc
     aacctagtaa cgccttncag gctccggcga agagaagaat agcttagcag agctattttc
     attttcggga gacgagatca agcagatcaa cggtcgtcaa gagacctacg agactgagga
     atccgctctt ggctccacgc gactatatat ttgtctctaa ttgtactttg acatgctcct
     cttctttact ctgatagctt gactatgaaa attccgtcac cagcncctgg gttcgcaaag
     ataattgcat gtttcttcct tgaactctca agcctacagg acacacattc atcgtaggta
     taaacctcga aatcanttcc tactaagatg gtatacaata gtaaccatgc atggttgcct
     agtgaatgct ccgtaacacc caatacgccg gccgaaactt ttttacaact ctcctatgag
     tcgtttaccc agaatgcaca ggtacacttg tttagaggta atccttcttt ctagagcttg
     gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacga attgcggccg
//