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ID   PWT551     preliminary; circular DNA; SYN; 3773 BP.
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AC   L09154; VB0088; IG0124;
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DT   02-NOV-1992 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli plasmid vector pWT551 - complete.
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KW   cloning vector.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RP   1-3773
RC   pWT551
RA   Gilbert W.;
RT   "Obtained from VecBase 3.0";
RL   Unpublished (1991).
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RN   [2]
RC   pWT221 from pAT153 & pWT121
RC   pWT501 from pAT153 & pWT221
RC   pWT511 from pWT501
RC   pWT550 from pAT153 & pWT501
RC   pWT551 from pWT550
RC   pWT551-2P2, pWT551-3P3 from pWT551
RC   pWT570 from pAT153 & pWT501
RC   pWT571 from pWT570
RC   pMN39-1 from pWT501 & human beta-interferon
RA   Tacon W., Bonass W.A., Jenkins B., Emtage J.S.;
RT   "Expression plasmid vectors containing Escherichia coli
RT   tryptophan promoter transcriptional units lacking the attenuator";
RL   Gene 23:255-265(1983).
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CC   Assembled from pAT153 and GenBank:EcoTrpPO according to the
CC   publication cited by F. Pfeiffer.
CC   There is confliciting information on the structure of pAT153.
CC   some 200 bases may be erraneuosly missing from this sequence.
CC
CC   The construction of three different types of trp promoter
CC   expression plasmids (pWT511, pWT551 and pWT571) is described.
CC   The Tc-resistance gene is under the control of the Trp-operator.
CC   In pWT511, the HindIII site is located several codons after
CC   the ATG and allows production of a fusion protein with few
CC   N-terminal amino acids from the Trp leader peptide.
CC
CC   The bom or Mob site is lost.
CC   NM (pWT551)
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 (pAT153)(EcoTrpPO)(EcoTrp)
CC   BR (pBR327)(pWT511)(pWT571)
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pBR322
FT                   2. ptrp ED5-1, E. coli trp operon
FT                   -> pWT101
FT                   1. pWT101
FT                   -> pWT111
FT                   1. pWT111
FT                   -> pWT121
FT                   1. pAT153 3658bp
FT                   2. pWT121, E. coli trp operon
FT                   -> pWT221
FT                   1. pAT153 HindIII 3658bp 30..30
FT                   2. pWT221 HhaI-HhaI 139bp, E. coli trp operon
FT                   DNA polymerase I:DNA polymerase I
FT                   HindIII linker 10bp ccaagcttgg:HindIII linker 10bp
FT                   \ ccaagcttgg
FT                   -> pWT501
FT                   1. pAT153 HindIII 3658bp 30..30
FT                   2. pWT501 HaeIII-HaeIII 137bp, trp promoter
FT                   HaeIII-TaqI 99bp, trp promoter
FT                   :DNA polymerase I
FT                   HindIII linker 10bp ccaagcttgg:HindIII linker 10bp
FT                   \ ccaagcttgg
FT                   -> pWT550 [unique HindIII]"
FT                   1. pWT550 HindIII, 5' to trp promoter
FT                   DNA polymerase I
FT                   -> pWT551 3773bp [unique HindIII]"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   misc_feature    1..10
FT                   /note="destroyed HindIII-linker"
FT   promoter        12..113
FT                   /note="PRO E. coli trp gene, from 99 bp HaeIII-TaqI
FT                   fragment from the trp operon"
FT   RBS             12..113
FT                   /note="RBS E. coli trp gene, from 99 bp HaeIII-TaqI
FT                   fragment from the trp operon"
FT   CDS             173..1363
FT                   /note="ANT E. coli tetracycline resistance gene (tet)"
FT   CDS             complement(2680..3468)
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
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SQ   Sequence 3773 BP; 850 A; 1035 C; 976 G; 912 T; 0 other;
     aagctagctt ggccgacatc acaacggttc tggcaaatat tctgaaatga gctgttgaca
     attaatcatc gaactagtta actagtacgc aagttcacgt aaaaagggta tcgccaagct
     ttaatgcggt agtttatcac agttaaattg ctaacgcagt caggcaccgt gtatgaaatc
     taacaatgcg ctcatcgtca tcctcggcac cgtcaccctg gatgctgtag gcataggctt
     ggttatgccg gtactgccgg gcctcttgcg ggatatcgtc cattccgaca gcatcgccag
     tcactatggc gtgctgctag cgctatatgc gttgatgcaa tttctatgcg cacccgttct
     cggagcactg tccgaccgct ttggccgccg cccagtcctg ctcgcttcgc tacttggagc
     cactatcgac tacgcgatca tggcgaccac acccgtcctg tggatcctct acgccggacg
     catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat
     caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg
     tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt
     ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga
     gtcgcataag ggagagcgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
     ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
     actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc gctttcgctg
     gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca
     agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg
     catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag gctggatggc
     cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat
     gctgtccagg caggtagatg acgaccatca gggacagctt caaggatcgc tcgcggctct
     taccagccta acttcgatca ctggaccgct gatcgtcacg gcgatttatg ccgcctcggc
     gagcacatgg aacgggttgg catggattgt aggcgccgcc ctataccttg tctgcctccc
     cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc tgaatggaag ccggcggcac
     ctcgctaacg gattcaccac tccaagaatt ggagccaatc aattcttgcg gagaactgtg
     aatgcgcaaa ccaacccttg gcagaacata tccatcgcgt ccgccatctc cagcagccgc
     acgcggcgca tctcgggcag cgttgggtcc tggccacggg tgcgcatgat cgtgctcctg
     tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg
     atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca
     tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgctcttc
     cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat
     cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga
     taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     cacgaggccc tttcgtcttc aagaattctc atgtttgaca gcttatcatc gat
//