LOCUS       pACYCDuet-1.        4008 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector with a p15A origin for the co-expression of two 
            genes.
ACCESSION   .
VERSION     .
KEYWORDS    pACYCDuet-1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4008)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4008)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             42..64
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             71..73
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             83..100
                     /label=6xHis
                     /note="6xHis affinity tag"
     promoter        214..232
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    233..257
                     /label=lac repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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             286..291
                     /note="ribosome binding site"
     CDS             300..302
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             366..410
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     terminator      462..509
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(732..1388)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(1389..1491)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(2017..2561)
                     /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."
     protein_bind    complement(2752..2773)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2789..3868)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(3869..3946)
                     /label=lacI promoter
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
      121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag
      181 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt
      241 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata
      301 tggcagatct caattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt
      361 ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg
      421 cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg
      481 cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg
      541 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
      601 aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc tgccattcat
      661 ccgcttatta tcacttattc aggcgtagca ccaggcgttt aagggcacca ataactgcct
      721 taaaaaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
      781 ctgccgacat ggaagccatc acagacggca tgatgaacct gaatcgccag cggcatcagc
      841 accttgtcgc cttgcgtata atatttgccc atagtgaaaa cgggggcgaa gaagttgtcc
      901 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa
      961 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
     1021 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
     1081 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
     1141 accagctcac cgtctttcat tgccatacgg aactccggat gagcattcat caggcgggca
     1201 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
     1261 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
     1321 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
     1381 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
     1441 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat
     1501 tttcgccaaa agttggccca gggcttcccg gtatcaacag ggacaccagg atttatttat
     1561 tctgcgaagt gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct
     1621 gccaacttac tgatttagtg tatgatggtg tttttgaggt gctccagtgg cttctgtttc
     1681 tatcagctgt ccctcctgtt cagctactga cggggtggtg cgtaacggca aaagcaccgc
     1741 cggacatcag cgctagcgga gtgtatactg gcttactatg ttggcactga tgagggtgtc
     1801 agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt
     1861 gatacaggat atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc
     1921 ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac
     1981 ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga
     2041 caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag
     2101 ataccaggcg tttcccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt
     2161 accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc
     2221 cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg
     2281 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc
     2341 actggcagca gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt
     2401 aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt
     2461 tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt
     2521 tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc
     2581 agataaaata tttctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca
     2641 gccccatacg atataagttg taattctcat gttagtcatg ccccgcgccc accggaagga
     2701 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     2761 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2821 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     2881 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     2941 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     3001 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     3061 ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     3121 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     3181 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     3241 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     3301 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     3361 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     3421 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     3481 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     3541 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     3601 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     3661 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     3721 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     3781 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     3841 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     3901 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
     3961 tctcccttat gcgactcctg cattaggaaa ttaatacgac tcactata
//
