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ID   PWT511     preliminary; circular DNA; SYN; 3809 BP.
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AC   L09153; A15710; A15711; A15712; A15713; A15714; VB0087; IG0123;
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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 pWT511 - 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   1-3809
RC   pWT511
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RL   Unpublished (1991).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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RN   [3]
RC   pWT101, pWT102 from pBR322 & ptrp ED5-1
RC   pWT111 from pWT101
RC   pWT121 from pWT111
RC   pWT131 from pWT121
RC   ptrpE2-1 from pBR322 & trp gene
RA   Tacon W., Carey N., Emtage J.S.;
RT   "The construction and characterization of plasmid vectors suitable
RT   for the expression of all DNA phases under the control of the
RT   E. coli tryptophan promoter";
RL   Mol. Gen. Genet. 177:427-438(1980).
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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 (pWT511)
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)(pWT551)(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. E. coli trp operon
FT                   -> ptrp ED5-1
FT                   1. pBR322 HindIII 4361bp 30..30
FT                   bacterial alkaline phosphatase
FT                   2. ptrp ED5-1 HinfI-HinfI 497bp, E. coli trp promoter
FT                   Klenow:Klenow
FT                   HindIII linker 10bp ccaagcttgg:HindIII linker 10bp
FT                   \ ccaagcttgg
FT                   -> pWT101 4874bp
FT                   1. pWT101 EcoRI
FT                   exonuclease III
FT                   Klenow:Klenow
FT                   -> pWT111 4823bp [unique HindIII]
FT                   1. pWT111 HindIII
FT                   Klenow
FT                   HindIII linker 10bp ccaagcttgg
FT                   -> pWT121 4837bp
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. pWT501 HindIII, 5' to trp promoter
FT                   DNA polymerase I
FT                   -> pWT511 3809bp [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..150
FT                   /note="PRO E. coli trp gene, from 139 bp HhaI-HhaI
FT                   fragment from the trp operon"
FT   RBS             12..150
FT                   /note="RBS E. coli trp gene, from 139 bp HhaI-HhaI
FT                   fragment from the trp operon"
FT   CDS             209..1399
FT                   /note="ANT E. coli tetracycline resistance gene (tet)"
FT   CDS             complement(2716..3504)
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
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SQ   Sequence 3809 BP; 862 A; 1039 C; 986 G; 922 T; 0 other;
     aagctagctt ggccgacatc acaacggttc tggcaaatat tctgaaatga gctgttgaca
     attaatcatc gaactagtta actagtacgc aagttcacgt aaaaagggta tcgacaatga
     aagcaatttt cgtactgaaa ggttggtggc caagctttaa tgcggtagtt tatcacagtt
     aaattgctaa cgcagtcagg caccgtgtat gaaatctaac aatgcgctca tcgtcatcct
     cggcaccgtc accctggatg ctgtaggcat aggcttggtt atgccggtac tgccgggcct
     cttgcgggat atcgtccatt ccgacagcat cgccagtcac tatggcgtgc tgctagcgct
     atatgcgttg atgcaatttc tatgcgcacc cgttctcgga gcactgtccg accgctttgg
     ccgccgccca gtcctgctcg cttcgctact tggagccact atcgactacg cgatcatggc
     gaccacaccc gtcctgtgga tcctctacgc cggacgcatc gtggccggca tcaccggcgc
     cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg
     ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg
     gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg
     cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgacc
     gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc
     gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac
     caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg cgctgggcta
     cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga ttcttctcgc
     ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgacga
     ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agcctaactt cgatcactgg
     accgctgatc gtcacggcga tttatgccgc ctcggcgagc acatggaacg ggttggcatg
     gattgtaggc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag
     ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt caccactcca
     agaattggag ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa cccttggcag
     aacatatcca tcgcgtccgc catctccagc agccgcacgc ggcgcatctc gggcagcgtt
     gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg
     cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac
     tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg tttccgtgtt
     tcgtaaagtc tggaaacgcg gaagtcagcg ctcttccgct tcctcgctca ctgactcgct
     gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
     tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
     tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga
     attctcatgt ttgacagctt atcatcgat
//