LOCUS       40924_29531        4886 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Plasmid for expressing high levels of T7 lysozyme, an inhibitor of 
            T7 RNA polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4886)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4886)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Expression of the T7 lysozyme gene is driven by the tet promoter.
FEATURES             Location/Qualifiers
     source          1..4886
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(220..322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(848..1392)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     promoter        1504..1532
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             1918..2370
                     /codon_start=1
                     /label=T7 lysozyme
                     /note="lysozyme from bacteriophage T7"
                     /translation="MARVQFKQRESTDAIFVHCSATKPSQNVGVREIRQWHKEQGWLDV
                     GYHFIIKRDGTVEAGRDEMAVGSHAKGYNHNSIGVCLVGGIDDKGKFDANFTPAQMQSL
                     RSLLVTLLAKYEGAGLRAHHEVAPKACPSFDLKRWWEKNELVTSDRG"
     promoter        2371..2404
                     /label=T7 Phi-3.8 promoter
                     /note="weak promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(join(4449..4886,1..219))
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
       61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
      181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
      241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
      301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
      361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
      421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
      481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
      541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
      601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
      841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttcccctg gcggctccct
      961 cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg
     1021 tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact
     1081 gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg
     1141 agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta
     1201 gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt
     1261 gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc
     1321 gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca
     1381 aaacgatctc aagaagatca tcttattaat cagataaaat atttctagat ttcagtgcaa
     1441 tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca
     1501 tgtttgacag cttatcatcg ataagcttta atgcggtagt ttatcacagt taaattgcta
     1561 acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc atcgtcatcc tcggcaccgt
     1621 caccctggat gctgtaggca taggcttggt tatgccggta ctgccgggcc tcttgcggga
     1681 tatcgtccat tccgacagca tcgccagtca ctatggcgtg ctgctagcgc tatatgcgtt
     1741 gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc gaccgctttg gccgccgccc
     1801 agtcctgctc gcttcgctac ttggagccac tatcgactac gcgatcatgg cgaccacacc
     1861 cgtcctgtgg atccgggtcc cctttgatag attaaaaagg aaaggaggaa agaaataatg
     1921 gctcgtgtac agtttaaaca acgtgaatct actgacgcaa tctttgttca ctgctcggct
     1981 accaagccaa gtcagaatgt tggtgtccgt gagattcgcc agtggcacaa agagcagggt
     2041 tggctcgatg tgggatacca ctttatcatc aagcgagacg gtactgtgga ggcaggacga
     2101 gatgagatgg ctgtaggctc tcacgctaag ggttacaacc acaactctat cggcgtctgc
     2161 cttgttggtg gtatcgacga taaaggtaag ttcgacgcta actttacgcc agcccaaatg
     2221 caatcccttc gctcactgct tgtcacactg ctggctaagt acgaaggcgc tggtcttcgc
     2281 gcccatcatg aggtggcgcc gaaggcttgc ccttcgttcg accttaagcg ttggtgggag
     2341 aagaacgaac tggtcacttc tgaccgtgga taattaattg aactcactaa agggagacca
     2401 cagcggtttc cctttgttcg cattggaggt caaataatgc gcaagtctta taaacaattc
     2461 tataaggctc cgaggaggca tatccaagtg tgggaggcag ccaatgggcc ggatcctcta
     2521 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat
     2581 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt
     2641 cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca
     2701 tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct
     2761 aatgcaggag tcgcataagg gagagcgtcg accgatgccc ttgagagcct tcaacccagt
     2821 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     2881 tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg
     2941 ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc
     3001 cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat
     3061 tatcgccggc atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg
     3121 ctggatggcc ttccccatta tgattcttct cgcttccggc ggcatcggga tgcccgcgtt
     3181 gcaggccatg ctgtccaggc aggtagatga cgaccatcag ggacagcttc aaggatcgct
     3241 cgcggctctt accagcctaa cttcgatcac tggaccgctg atcgtcacgg cgatttatgc
     3301 cgcctcggcg agcacatgga acgggttggc atggattgta ggcgccgccc tataccttgt
     3361 ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc
     3421 cggcggcacc tcgctaacgg attcaccact ccaagaattg gagccaatca attcttgcgg
     3481 agaactgtga atgcgcaaac caacccttgg cagaacatat ccatcgcgtc cgccatctcc
     3541 agcagccgca cgcggcgcat ctcgggcagc gttgggtcct ggccacgggt gcgcatgatc
     3601 gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat
     3661 gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga
     3721 gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtcc
     3781 cctacgtgct gctgaagttg cccgcaacag agagtggaac caaccggtga taccacgata
     3841 ctatgactga gagtcaacgc catgagcggc ctcatttctt attctgagtt acaacagtcc
     3901 gcaccgctgt ccggtagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     3961 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgcc caacagtccc
     4021 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgccctgcac cattatgttc
     4081 cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg
     4141 aagcgctaac cgtttttatc aggctctggg aggcagaata aatgatcata tcgtcaatta
     4201 ttacctccac ggggagagcc tgagcaaact ggcctcaggc atttgagaag cacacggtca
     4261 cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac
     4321 gaccctgccc tgaaccgacg accgggtcga atttgctttc gaatttctgc cattcatccg
     4381 cttattatca cttattcagg cgtagcacca ggcgtttaag ggcaccaata actgccttaa
     4441 aaaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg
     4501 ccgacatgga agccatcaca gacggcatga tgaacctgaa tcgccagcgg catcagcacc
     4561 ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata
     4621 ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac
     4681 atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct
     4741 tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa
     4801 aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc
     4861 agctcaccgt ctttcattgc catacg
//
