LOCUS       V013182                 7297 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013182
VERSION     V013182
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7297)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(140..157)
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             complement(164..2227)
                     /gene="ppk"
                     /label="Polyphosphate kinase"
                     /note="Polyphosphate kinase from Escherichia coli O157:H7.
                     Accession#: P0A7B2"
     RBS             complement(2234..2256)
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(2271..2295)
                     /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)."
     promoter        complement(2296..2314)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2623..2700
                     /label="lacI promoter"
     CDS             2701..3780
                     /label="lacI"
                     /note="lac repressor"
     protein_bind    3796..3817
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             4592..4780
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    4885..5027
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5213..5801)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             5923..6735
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(6831..7286)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagttcaggt tgttcaagag
      181 atttgatgta atcgtagata gcaagttgag cacgaacttt acgacggttg ccacgaggaa
      241 cgtaacggtt agaaagttct ttatcgatgt aacgagcttt aactgtatca gagaaaagga
      301 tatcgatgat atcaagaaca cgttgtttaa gacgaggatc aagaagaggt gtagcaactt
      361 cgatacggta atcgatgtta cgtgtcatcc aatcagcaga agaaaggtaa acttttttat
      421 cgccgccgtt ttcgaagatg taaacacgat catgttcaag gtaacgatca acgatagaga
      481 tagcacggat gttatcagag atgccttcaa ggttagggat aagagagcac atgccacgaa
      541 caagaaggtt aacaggaacg ccagaagaag aagcagcgta aagacgatca acaaggcctt
      601 tatcaacaag gttgttaagt ttaagtgtga tgccagaagg aaggccttgt tgagcgttag
      661 cgatttcacg atcaaccatt tcgtaaagaa gacgacgaga gttttgagga gaaaccataa
      721 ggtaatcgaa tgtaacagga cggtaagggt tttcgatgaa gttgaaaaca cgacgaactt
      781 cgtttgtgat acgagcatca gctgtaagaa gagagtaatc tgtgtaaaga cgagctgttt
      841 tttcgttgaa gttgcctgtg ccgatatgag cgtaacgaac aacttcgccg ttttctttac
      901 gagagataag gaaaagttta gcatggattt taaggccagg agcagagaag ataacatgaa
      961 cgccagcttc tgtaagacgt ttagcccaat ggatgttagc ttcttcatcg aaacgagctt
     1021 gaagttcaac aacaactgta acttttttgc cgttatgagc agcatggatc atagaatcga
     1081 tgatacgaga atctttagca acacggtaga tgttgatttt gatagcaaga acagaaggat
     1141 cgaaagaagc ttgacgaaga agttcaagaa catgttcgaa tgtatggtaa gggtagtaaa
     1201 gaagaacatc acgttcacgg atagcatcga agccgttacg gaattgagct ttatcgaacc
     1261 agatatgacg aagacgagga agaggtttgt taacaaggtt agctttgcca acgttaggga
     1321 agttgatgaa atctttgaag ttatggtaac ggccgccagg aacgatagaa tcgtaacgag
     1381 agattgtaag tttttcacga agaacttcaa caagagcgtt aggcatatca cgttggtaaa
     1441 cgaaacgaac aggttcagct gtaagacgtt gtttaagaga agaagacata agttccataa
     1501 gagaagcttc catttcatga acaagatcgt attcagcatc acgtgtcatt ttcatagagt
     1561 aagcgttaag agcatcgtaa tcgaagaagc ctttgaagat atcatcaagg cagtaacgaa
     1621 ggatgttatc aagaaggatc ataggtttac gacgacgagg agcttcagga ggaaggttaa
     1681 cgaaacgagg aactttatca gaagggattt caagaagagc gtaacggatt gtatcgccac
     1741 ggatgatttc aacagcaagg tatgtgtaat catctttaag gaattgaaca agatctgtat
     1801 cagggttgat aaggataggt gtgatatgtt gacgaaggta ttgtttgaag taatgacgaa
     1861 gccagttttg ttggttaaca gaaagttgac gttcgttgat aaggaagatt tggttacgag
     1921 ccatttcaag aagaagttcg ttgtaaaggc catcgaattc ttgatcagct ttaagaacac
     1981 gagattggat tttgccaaga agatgacgag aatgagagtt agagccttgt tcttcagaga
     2041 tgatgatacg acgtttaagt tcagcgaaac gaactttgta gaattcatca aggttgttag
     2101 agtagatgcc aaggaaacgc atacgttcga taagagggtt agatttatca gcagcttctt
     2161 gaagaacacg ttcgttgaaa gaaagccaag aaagttcttt ttcgatgtaa agtttttctt
     2221 ggcccatggt atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta
     2281 tccgctcaca attcccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc
     2341 tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct
     2401 atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt
     2461 gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct
     2521 tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct
     2581 tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc
     2641 aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat
     2701 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
     2761 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
     2821 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
     2881 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
     2941 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
     3001 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt
     3061 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc
     3121 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
     3181 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag
     3241 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
     3301 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
     3361 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
     3421 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
     3481 gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca
     3541 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
     3601 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc
     3661 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc
     3721 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     3781 tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat
     3841 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     3901 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     3961 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct
     4021 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa
     4081 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccccacggg tgcgcatgat
     4141 cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa
     4201 tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg
     4261 agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc
     4321 agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg
     4381 aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc
     4441 ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc
     4501 atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat
     4561 gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa
     4621 catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga
     4681 cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg
     4741 cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     4801 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     4861 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     4921 gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatatgc
     4981 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     5041 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5101 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5161 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5221 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5281 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5341 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5401 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5461 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5521 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5581 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5641 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5701 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5761 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5821 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgaaca
     5881 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     5941 gaaacgtctt gctctaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     6001 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag
     6061 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     6121 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     6181 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg gaaaacagca
     6241 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     6301 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     6361 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     6421 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg
     6481 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     6541 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     6601 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     6661 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     6721 ctcgatgagt ttttctaaga attaattcat gagcggatac atatttgaat gtatttagaa
     6781 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg aaattgtaaa
     6841 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     6901 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     6961 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     7021 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     7081 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     7141 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     7201 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     7261 gcttaatgcg ccgctacagg gcgcgtccca ttcgcca
//
