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ID   PAN562H    preliminary; circular DNA; SYN; 6114 BP.
XX
AC   Z32690;
XX
DT   01-JUL-1994 (Rel. 9, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE   E. coli plasmid vector pAN56-2H - complete.
XX
KW   cloning vector.
XX
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
XX
RN   [1]
RP   1-850
RC   pABpyrGNot from pBluescript & linker & pAB94-11, pyrG* gene
RC   pAN52-7NotI from pAN52-1NotI & A.niger glaA gene promoter
RC   pGUS54 from pABpyrGNot & pNOM102, E. coli uidA gene
RC   pGUS64 from pAN52-7NotI & pNOM102, E. coli uidA gene
RC   plasmid from pMTL25P & pAN52-6NotI, glaA gene promoter
RC   pAN4-1 from pAN4
RC   pAN4-cos1 from pAN4 & lambda, cos
RC   pAB6-15 from plasmid & pAN4-1
RC   pAB6-18 from plasmid & pAN4-cos1
RA   Verdoes J.C., Punt P.J., Stouthamer A.H., Van Den Hondel C.A.M.J.J.;
RT   "The effect of multiple copies of the upstream region of the
RT   Aspergillus niger glucoamylase-encoding gene on expression";
RL   Gene 145:179-187(1994).
XX
RN   [2]
RP   1-6114
RC   pAN56-2H, 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RL   Unpublished (1994).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   1-6114
RC   pAN56-2H
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
CC   NCBI gi: 474930
CC   NM (pAN56-2H)
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 (pAN56-1)
CC   BR ()
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pTZ19R 2863bp
FT                   2. Aspergillus niger, gLAA promoter
FT                   3. Aspergillus niger, GLAA2 gene
FT                   4. Aspergillus nidulans, trpC gene terminator
FT                   5. lambda, cos
FT                   -> pAN56-1 6100bp
FT                   1. pAN56-1
FT                   -> pAN56-2H 6114bp"
FT   promoter        36..781
FT                   /note="PRO A. niger gLAA gene;
FT                   EXPERIMENTAL [1] [2]"
FT   misc_feature    2134..2134
FT                   /note="A. niger gLAA gene major
FT                   transcription start site; EXPERIMENTAL [1]"
FT   misc_RNA        851..2719
FT                   /note="A.niger glucoamylase gene (encoding GII form
FT                   only) GLAA2 gene; EXPERIMENTAL [2];
FT                   #XOO712 with several modifications; see other
FT                   features; this region contains 4 introns"
FT   sig_peptide     815..904
FT                   /note="glaa secretion signal sequence
FT                   identical to that of the published glaa gene;
FT                   EXPERIMENTAL [2]"
FT   misc_feature    905..922
FT                   /note="PRO_PEPTIDE sequence for putative propeptide,
FT                   contains dibasic processing site (LYS-ARG) [2]"
FT   misc_feature    925..2617
FT                   /note="this region was taken from accession X00712.
FT                   GLAA gene present in PAN56-1 is from another A.niger
FT                   (N402) and could therefore have minor deviations from
FT                   the sequence given here [2]"
FT   terminator      2751..3474
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 3011..3021
FT                   /note="replace(3011..3021,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 3220..3228
FT                   /note="replace(3220..3228,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 3308..3318
FT                   /note="replace(3308..3318,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        3345..3352
FT                   /note="replace(3345..3352,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      3001..3011
FT                   /note="partial"
FT   polyA_site      3274..3284
FT                   /note="partial"
FT   misc_feature    3474..6114
FT                   /note="from pUC18 HindIII-EcoRI [2]"
FT   misc_feature    1..6114
FT                   /note="fungal expression/secretion vector pAN56-1;
FT                   EXPERIMENTAL [2]"
XX
SQ   Sequence 6114 BP; 1429 A; 1617 C; 1545 G; 1520 T; 3 other;
     gcggccgctg tgcagaattc gagctcggta cccggggatc cgaactccaa ccggggggag
     tacattgagt ggccgcagtg gaaggaatcg cggcagttga tgaatttcgg agcgaacgac
     gccagtctcc ttacggatga tttccgcaac gggacatatg agttcatcct gcagaatacc
     gcggcgttcc acatctgatg ccattggcgg aggggtccgg acggtcagga acttagcctt
     atgagatgaa tgatggacgt gtctggcctc ggaaaaggat atatggggat cataatagta
     ctagccatat taatgaaggg catataccac gcgttggacc tgcgttatag cttcccgtta
     gttatagtac catcgttata ccagccaatc aagtcaccac gcacgaccgg ggacggcgaa
     tccccgggaa ttgaaagaaa ttgcatccta ggccagtgag gccagcgatt ggccacctct
     ccaagcacac agggccattc tgcagcgctg gtggattcat cgcaatttcc cccggcccgg
     cccgacaccg ctataggctg gttctcccac accatcggag attcgtcgcc taatgtctcg
     tccgttcaca agctgaagag cttgaagtgg cgagatgtct ctgcaggaat tcaagctaga
     tgctaagcga tattgcatgg caatatgtgt tgatgcatgt gcttcttcct tcagcttccc
     ctcgtgcaga tgaggtttgg ctataaattg aagtggttgg tcggggttcc gtgaggggct
     gaagtgcttc ctccctttta gacgcaactg agagcctgag cttcatcccc agcatcatta
     cacctcagca atgtcgttcc gatctcttct cgccctgagc ggccttgtct gcacagggtt
     ggcaaatgtg atttccaagc gcgcgacctt ggattcatgg ttgagcaacg aagcgaccgt
     ggctcgtact gccatcctga ataacatcgg ggcggacggt gcttgggtgt cgggcgcgga
     ctctggcatt gtcgttgcta gtcccagcac ggataacccg gactgtatgt ttcgagctca
     gatttagtat gagtgtgtca ttgattgatt gatgctgact ggcgtgtcgt ttgttgtaga
     cttctacacc tggactcgcg actctggtct cgtcctcaag accctcgtcg atctcttccg
     aaatggagat accagtctcc tctccaccat tgagaactac atctccgccc aggcaattgt
     ccagggtatc agtaacccct ctggtgatct gtccagcggc gctggtctcg gtgaacccaa
     gttcaatgtc gatgagactg cctacactgg ttcttgggga cggccgcagc gagatggtcc
     ggctctgaga gcaactgcta tgatcggctt cgggcagtgg ctgcttgtat gttctccacc
     cccttgcgtc tgatctgtga catatgtagc tgactggtca ggacaatggc tacaccagca
     ccgcaacgga cattgtttgg cccctcgtta ggaacgacct gtcgtatgtg gctcaatact
     ggaaccagac aggatatggt gtgtttgttt tattttaaat ttccaaagat gcgccagcag
     agctaacccg cgatcgcaga tctctgggaa gaagtcaatg gctcgtcttt ctttacgatt
     gctgtgcaac accgcgccct tgtcgaaggt agtgccttcg cgacggccgt cggctcgtcc
     tgctcctggt gtgattctca ggcacccgaa attctctgct acctgcagtc cttctggacc
     ggcagcttca ttctggccaa cttcgatagc agccgttccg gcaaggacgc aaacaccctc
     ctgggaagca tccacacctt tgatcctgag gccgcatgcg acgactccac cttccagccc
     tgctccccgc gcgcgctcgc caaccacaag gaggttgtag actctttccg ctcaatctat
     accctcaacg atggtctcag tgacagcgag gctgttgcgg tgggtcggta ccctgaggac
     acgtactaca acggcaaccc gtggttcctg tgcaccttgg ctgccgcaga gcagttgtac
     gatgctctat accagtggga caagcagggg tcgttggagg tcacagatgt gtcgctggac
     ttcttcaagg cactgtacag cgatgctgct actggcacct actcttcgtc cagttcgact
     tatagtagca ttgtagatgc cgtgaagact ttcgccgatg gcttcgtctc tattgtggta
     agtctacgct agacaagcgc tcatgttgac agagggtgcg tactaacaga agtaggaaac
     tcacgccgca agcaacggct ccatgtccga gcaatacgac aagtctgatg gcgagcagct
     ttccgctcgc gacctgacct ggtcttatgc tgctctgctg accgccaaca accgtcgtaa
     ctccgtcgtg cctgcttctt ggggcgagac ctctgccagc agcgtgcccg gcacctgtgc
     ggccacatct gccattggta cctacagcag tgtgactgtc acctcgtggc cgagtatcgt
     ggctactggc ggcaccacta cgacggctac ccccactgga tccggcagcg tgacctcgac
     cagcaagacc accgcgactg ctagcaagac cagcaccagt acgtcatcaa cctcctgtac
     cactcccacg gcgccgtaaa gatctgctag aggatctggt tacagcttga gatccactta
     acgttactga aatcatcaaa cagcttgacg aatctggata taagatcgtt ggtgtcgatg
     tcagctccgg agttgagaca aatggtgttc aggatctcga taagatacgt tcatttgtcc
     aagcagcaaa gagtgccttc tagtgattta atagctccat gtcaacaaga ataaaacgcg
     ttttcgggtt tacctcttcc agatacagct catctgcaat gcattaatgc attgactgca
     acctagtaac gccttncagg ctccggcgaa gagaagaata gcttagcaga gctattttca
     ttttcgggag acgagatcaa gcagatcaac ggtcgtcaag agacctacga gactgaggaa
     tccgctcttg gctccacgcg actatatatt tgtctctaat tgtactttga catgctcctc
     ttctttactc tgatagcttg actatgaaaa ttccgtcacc agcncctggg ttcgcaaaga
     taattgcatg tttcttcctt gaactctcaa gcctacagga cacacattca tcgtaggtat
     aaacctcgaa atcanttcct actaagatgg tatacaatag taaccatgca tggttgccta
     gtgaatgctc cgtaacaccc aatacgccgg ccgaaacttt tttacaactc tcctatgagt
     cgtttaccca gaatgcacag gtacacttgt ttagaggtaa tccttctttc tagagcttgg
     cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     gcccttccca acagttgcgc agcctgaatg gcgaatggcg cgcctgatgc ggtattttct
     ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg aatt
//