LOCUS       V014059                 9549 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014059
VERSION     V014059
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9549)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2265..4004
                     /gene="mcjD"
                     /label="Microcin-J25 export ATP-binding/permease protein
                     McjD"
                     /note="Microcin-J25 export ATP-binding/permease protein
                     McjD from Escherichia coli. Accession#: Q9X2W0"
     promoter        4079..4097
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4098..4122
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             4537..4707
                     /gene="mcjA"
                     /label="Microcin J25"
                     /note="Microcin J25 from Escherichia coli. Accession#:
                     Q9X2V7"
     CDS             4723..4767
                     /label="S-Tag"
                     /note="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
     terminator      4819..4866
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     rep_origin      4903..5358
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(5476..6333)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6334..6426)
                     /label="AmpR promoter"
     rep_origin      6507..7095
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(7281..7423)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(7528..7716)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(8254..8275)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(8291..9370)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(9371..9448)
                     /label="lacI promoter"
ORIGIN
        1 ctctttttga tgcaatccgc tttgcttctg actataatag tcagggtaat tgagtgtaaa
       61 ggcataacta caggagggag tgtgcaaaat gatccgttac tgcttaacca gttatagaga
      121 ggatcttgtt atcctggata taattaatga tagtttcagc atagtgcctg acgcaggtag
      181 cttgctaaaa gaaagagata aattgcttaa agaattccca caactatctt acttttttga
      241 cagtgaatat catattggaa gtgtttctcg taatagtgac acttcttttc ttgaagaacg
      301 ctggtttcta ccagaacctg acaaaacatt atataagtgt tctctattta aacgatttat
      361 attattactc aaagtctttt actatagctg gaatattgaa aaaaaaggga tggcatggat
      421 tttcataagt aataaaaaag agaataggct atactccttg aatgaagagc atcttatccg
      481 gaaagaaatt agtaatcttt ccattatctt tcatcttaat atttttaaat ctgactgtct
      541 tacctattca tacgcactaa aaagaattct taattccaga aatattgatg ctcatcttgt
      601 tattggtgta aggacacaac ctttttatag ccactcttgg gtggaggttg ggggacaagt
      661 tatcaatgat gctcccaata tgcgggataa attatctgtt attgcagaga tatagttatg
      721 gaaatattta atgtcaagtt aaatgatact tcaattagaa ttattttctg taaaacgctt
      781 tctgccttcc ggacagaaaa taccatcgtt atgctcaaag gaaaagcagt ttcaaatggc
      841 aaacctgtat ccacagagga gattgccaga gtagtggaag aaaaaggtgt ttcagaagta
      901 atagaaaatt tagatggtgt tttctgtatc ctaatttatc attttaatga tctccttata
      961 gggaaaagca ttcaatcagg ccccgctcta ttttattgta aaaagaatat ggatattttt
     1021 gtttcggata aaatttctga tatcaaattt ttgaatccag atatgacatt cagtctaaat
     1081 ataacaatgg cagaacatta tctgtcagga aatcgaatag caacccagga atcactaatc
     1141 actggcattt acaaagtaaa taatggtgag tttataaaat ttaataatca gttgaaacct
     1201 gtgctacttc gtgatgagtt tagtattacc aaaaagaaca attcaactat cgacagtatc
     1261 attgataata ttgagatgat gcgggataat agaaaaatag ccctattatt ttccggagga
     1321 ttggattctg cattaatttt tcacacactt aaagaatcag gtaacaaatt ctgcgcttat
     1381 catttttttt ctgatgaatc tgatgacagt gaaaagtatt ttgctaagga atactgttca
     1441 aaatatggag ttgattttat atctgttaat aaaaacatca actttaatga aaaactttat
     1501 ttcaatttaa atcctaatag tccggacgaa atccctttga tatttgaaca gacagatgaa
     1561 gaaggtgaag gtcagccccc catagacgat gatttattat atctatgtgg tcacggtgga
     1621 gatcatattt tcggacaaaa tccttcagaa ctttttggca ttgatgcata tcgaagtcat
     1681 ggcttgatgt ttatgcataa aaaaatagta gaattttcca atctcaaggg aaagagatat
     1741 aaagatatca tattttcaaa tatttccgca ttcattaata catccaacgg atgttctcca
     1801 gcaaagcaag agcacgtatc agatatgaaa cttgcctctg ctcagttttt tgcaactgat
     1861 tatacaggaa aaattaataa actaactcca ttcctgcata aaaatattat ccagcattat
     1921 gctggcttac cagtttttag tctatttaac cagcactttg atcgttatcc cgttcgttat
     1981 gaagcgtttc aacgatttgg ttcagatatt ttctggaaaa aaaccaaacg gtcatcttca
     2041 tcatgactaa tattcagaat tctatccggt aaaaaggatg aactagtgaa tacaataaaa
     2101 cagtcaggat taattgaaat attaggcatt aaccatattg aattggaaag cattttgtat
     2161 gaaaatacga ctacacgtct gacaatggaa ctaccatata tacttaactt ataccgtctg
     2221 gcaaaattca ttcaacttca atccattgat tataaaggtt aattatggaa agaaaacaga
     2281 aaaactcatt atttaattat atttattcat taatggatgt aagaggtaaa tttttattct
     2341 tttccatgtt attcattaca tcattatcat cgataatcat atctatttca ccattgattc
     2401 ttgcaaagat tacagattta ctgtctggct cattgtcaaa ttttagttat gaatatctgg
     2461 ttttacttgc ctgtttatac atgttttgcg ttatatctaa taaagcaagt gtttttttat
     2521 ttatgatact gcaaagtagt ctacgtatta acatgcagaa aaaaatgtcg ctaaagtatt
     2581 tgagagaatt gtataacgaa aatataacta acttgagtaa aaataatgct ggatatacaa
     2641 cgcaaagtct taaccaggct tcaaatgaca tttatattct tgtgagaaat gtttcccaga
     2701 atatcctgtc acctgttata caacttattt ccactattgt tgttgtttta tctacgaagg
     2761 actggttttc tgccggtgtg ttttttctct atattctggt atttgtaatt tttaatacca
     2821 gactgactgg cagtttagcg tctctcagaa aacacagcat ggatatcact cttaactctt
     2881 atagtctgtt atctgatact gttgataaca tgatagcagc taaaaagaat aatgcattaa
     2941 gacttatttc tgaacgttat gaagatgctc tcactcagga aaacaatgct cagaaaaaat
     3001 actggttact cagttctaaa gttcttttat tgaactcttt acttgctgta atattatttg
     3061 gttctgtatt catatataat attttaggtg tgctgaatgg tgtagttagt atcggccact
     3121 tcattatgat tacatcatat atcattcttc tttcaacgcc agtggaaaat ataggggcat
     3181 tgctaagtga gatcaggcag tcaatgtcta gcctggcagg ttttattcaa cgtcatgccg
     3241 agaataaagc cacatctcct tcaatacctt ttctcaacat ggagcgaaaa ttaaacctgt
     3301 ccataagaga gctttcattt agctatagtg atgataaaaa aatacttaat tcagtcagtc
     3361 ttgacctttt taccggaaaa atgtattcat taaccggacc cagtggttca ggaaaatcca
     3421 cccttgtaaa aataatatca ggttactata aaaattactt tggagacatt tatctgaatg
     3481 atatatcctt acgtaatatc agtgatgagg atttgaatga tgctatttac tacctaacac
     3541 aagatgatta tatttttatg gatacactac gatttaatct ccggctcgca aattacgacg
     3601 cgtcagaaaa tgaaatattt aaagttctta aactggcaaa tctttctgtc gtcaacaatg
     3661 aaccagtgag tctggataca caccttataa acagaggcaa taactattca ggagggcaaa
     3721 aacaacgaat ttcgttagcg cgactgtttt tgagaaaacc tgcaataatt attattgatg
     3781 aagccacatc ggctctggat tatattaatg aatcagaaat tttatcatca ataagaactc
     3841 attttcctga tgcgttaatt ataaatatta gtcaccgaat aaatcttctg gagtgttccg
     3901 attgtgttta tgtattgaat gaaggaaata ttgttgcttc tggccatttc agggatttga
     3961 tggtcagcaa tgaatacata tcgggactgg cttctgttac tgaataaaag cttgcggccg
     4021 cataatgctt aagtcgaaca gaaagtaatc gtattgtaca cggccgcata atcgaaatta
     4081 atacgactca ctatagggga attgtgagcg gataacaatt ccccatctta gtatattagt
     4141 taagtataag aaggagatat acatatgacc atgtacgcaa cgcttgaaga agccattgac
     4201 gctgcacgcg aagaatttct tgcagacaac cccggcatcg acgccgaaga tgcgaatgtg
     4261 caacagttca atgcccaaaa atacgttttg caggacggcg acatcatgtg gcaagttgag
     4321 ttttttgccg acgaagggga agaaggtgaa tgtttaccta tgcttagcgg tgaagccgcg
     4381 caaagtgttt ttgatggcga ctatgatgag atagagatac gccaggagtg gcaggaagag
     4441 aatacattac atgaatggga cgagggggaa tttcagcttg agccaccgct ggataccgag
     4501 gaaggacgcg cagcagctga tgagtgggat gaacgtatta agcattttca ttttaataaa
     4561 ctgtcttctg gtaaaaaaaa taatgttcca tctcctgcaa agggggttat acaaataaaa
     4621 aaatcagcat cgcaactcac aaaaggtggt gcaggacatg tgcctgagta ttttgtgggg
     4681 attggtacac ctatatcttt ctatggctga ctcgagtctg gtaaagaaac cgctgctgcg
     4741 aaatttgaac gccagcacat ggactcgtct actagcgcag cttaattaac ctaggctgct
     4801 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     4861 tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg
     4921 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4981 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5041 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5101 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5161 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5221 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5281 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5341 tattaacgtt tacaatttct ggcggcacga tggcatgaga ttatcaaaaa ggatcttcac
     5401 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5461 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5521 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5581 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5641 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5701 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5761 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5821 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5881 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5941 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6001 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6061 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6121 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6181 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6241 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6301 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat catgattgaa
     6361 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6421 aacaaatagg tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6481 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6541 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6601 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     6661 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6721 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6781 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6841 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6901 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6961 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7021 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7081 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7141 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7201 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     7261 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     7321 tcacaccgca tatatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     7381 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     7441 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     7501 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     7561 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     7621 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     7681 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     7741 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     7801 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     7861 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     7921 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     7981 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     8041 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     8101 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     8161 gagcacgatc atgctagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     8221 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     8281 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     8341 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     8401 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     8461 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     8521 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa
     8581 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     8641 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     8701 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     8761 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     8821 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     8881 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     8941 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     9001 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     9061 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     9121 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     9181 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     9241 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     9301 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     9361 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     9421 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     9481 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     9541 ggtgcatgc
//
