LOCUS       V013296                 8033 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013296
VERSION     V013296
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8033)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(5..144)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(249..437)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(1715..1736)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(1752..2831)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(2832..2909)
                     /label="lacI promoter"
     promoter        3218..3236
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3237..3261
                     /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)."
     RBS             3276..3298
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3311..3337
                     /label="9xHis"
                     /note="9xHis affinity tag"
     CDS             3353..3364
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
     CDS             3368..5692
                     /gene="pol"
                     /label="DNA polymerase"
                     /note="DNA polymerase from Pyrococcus furiosus (strain ATCC
                     43587 / DSM 3638 / JCM 8422 / Vc1). Accession#: P61875"
     terminator      5765..5812
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     promoter        complement(5987..6015)
                     /label="tet promoter"
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        6128..6232
                     /label="AmpR promoter"
     CDS             6233..7090
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      7264..7852
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
       61 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
      121 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
      181 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
      241 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
      301 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
      361 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
      421 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
      481 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
      541 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
      601 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
      661 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
      721 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
      781 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
      841 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
      901 cgtggccagg acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac
      961 gcgatggata tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga
     1021 ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg
     1081 tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg
     1141 cggcgcctac aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg
     1201 tgacgatcag cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa
     1261 gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc
     1321 cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga
     1381 acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct
     1441 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     1501 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     1561 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     1621 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     1681 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     1741 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     1801 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     1861 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
     1921 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
     1981 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
     2041 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     2101 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     2161 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     2221 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     2281 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     2341 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     2401 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     2461 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     2521 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     2581 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     2641 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     2701 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     2761 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     2821 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     2881 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
     2941 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     3001 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     3061 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     3121 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     3181 tccggcgtag aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa
     3241 ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata
     3301 taccatgggc catcatcatc atcatcatca tcatcatcac agcagcggcc atatcgaagg
     3361 tcgtcatatg attttagatg tggattacat aactgaagaa ggaaaacctg ttattaggct
     3421 attcaaaaaa gagaacggaa aatttaagat agagcatgat agaactttta gaccatacat
     3481 ttacgctctt ctcagggatg attcaaagat tgaagaagtt aagaaaataa cgggggaaag
     3541 gcatggaaag attgtgagaa ttgttgatgt agagaaggtt gagaaaaagt ttctcggcaa
     3601 gcctattacc gtgtggaaac tttatttgga acatccccaa gatgttccca ctattagaga
     3661 aaaagttaga gaacatccag cagttgtgga catcttcgaa tacgatattc catttgcaaa
     3721 gagatacctc atcgacaaag gcctaatacc aatggagggg gaagaagagc taaagattct
     3781 tgccttcgat atagaaaccc tctatcacga aggagaagag tttggaaaag gcccaattat
     3841 aatgattagt tatgcagatg aaaatgaagc aaaggtgatt acttggaaaa acatagatct
     3901 tccatacgtt gaggttgtat caagcgagag agagatgata aagagatttc tcaggattat
     3961 cagggagaag gatcctgaca ttatagttac ttataatgga gactcattcg acttcccaca
     4021 tttagcgaaa agggcagaaa aacttgggat taaattaacc attggaagag atggaagcga
     4081 gcccaagatg cagagaatag gcgatatgac ggctgtagaa gtcaagggaa gaatacattt
     4141 cgacttgtat catgtaataa caaggacaat aaatctccca acatacacac tagaggctgt
     4201 atatgaagca atttttggaa agccaaagga gaaggtatac gccgacgaga tagcaaaagc
     4261 ctgggaaagt ggagagaacc ttgagagagt tgccaaatac tcgatggaag atgcaaaggc
     4321 aacttatgaa ctcgggaaag aattccttcc aatggaaatt cagctttcaa gattagttgg
     4381 acaaccttta tgggatgttt caaggtcaag cacagggaac cttgtagagt ggttcttact
     4441 taggaaagcc tacgaaagaa acgaagtagc tccaaacaag ccaagtgaag aggagtatca
     4501 aagaaggctc agggagagct acacaggtgg attcgttaaa gagccagaaa aggggttgtg
     4561 ggaaaacata gtatacctag attttagagc cctatatccc tcgattataa ttacccacaa
     4621 tgtttctccc gatactctaa atcttgaggg atgcaagaac tatgatatcg ctcctcaagt
     4681 aggccacaag ttctgcaagg acatccctgg ttttatacca agtctcttgg gacatttgtt
     4741 agaggaaaga cgaaagatta agacaaaaat gaaggaaact caagatccta tagaaaaaat
     4801 actccttgac tatagacaaa aagcgataaa actcttagca aattctttct acggatatta
     4861 tggctatgca aaagcaagat ggtactgtaa ggagtgtgct gagagcgtta ctgcctgggg
     4921 aagaaagtac atcgagttag tatggaagga gctcgaagaa aagcttggat ttaaagtcct
     4981 ctacattgac actgatggtc tctatgcaac tatcccagga ggagaaagtg aggaaataaa
     5041 gataaaggct ctagaatttg taaaatacat aaattcaaag ctccctggac tgctagagct
     5101 tgaatatgaa gggttttata agaggggatt cttcgttacg aagaagaggt atgcagtaat
     5161 agatgaagaa ggaaaagtca ttactcgtgg tttagagata gttaggagag attggagtga
     5221 aattgcaaaa gaaactcaag ctagagtttt ggagacaata ctaaaacacg gagatgttga
     5281 agaagctgtg agaatagtaa aagaagtaat acaaaagctt gccaattatg aaattccacc
     5341 agagaagctc gcaatatatg agcagataac aagaccatta catgagtata aggcgatagg
     5401 tcctcacgta gctgctgcaa agaaactagc tgctaaagga gttaaaataa agccaggaat
     5461 ggtaattgga tacatagtac ttagaggcga tggtccaatt agcaataggg caattctagc
     5521 tgaggaatac gatcccaaaa agcacaagta tgacgcagaa tattacattg agaaccaggt
     5581 tcttccagcg gtacttagga tattggaggg atttggatac agaaaggaag acctcagata
     5641 ccaaaagaca agacaagtcg gcctaacttc ctggcttaac attaaaaaat cctagctcga
     5701 ggatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     5761 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     5821 aggaactata tccggatatc ccgcaagagg cccggcagta ccggcataac caagcctatg
     5881 cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata
     5941 cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg
     6001 atgataagct gtcaaacatg agaattcttg aagacgaaag ggcctcgtga tacgcctatt
     6061 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     6121 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     6181 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     6241 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     6301 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     6361 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6421 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     6481 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6541 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     6601 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     6661 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6721 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     6781 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6841 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6901 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6961 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     7021 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     7081 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     7141 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     7201 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7261 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7321 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7381 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     7441 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7501 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7561 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7621 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     7681 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7741 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7801 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7861 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7921 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     7981 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aag
//
