LOCUS       pET-16b.        5711 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for inducible expression of N-terminally 
            10xHis-tagged proteins with a Factor Xa site.
ACCESSION   .
VERSION     .
KEYWORDS    pET-16b.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5711)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5711)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..38
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     terminator      complement(213..260)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    319..335
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(336..347)
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     CDS             complement(363..389)
                     /label=9xHis
                     /note="9xHis affinity tag"
     CDS             complement(393..395)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(402..424)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(439..463)
                     /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(464..482)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        791..868
                     /label=lacI promoter
     CDS             869..1948
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1964..1985
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3263..3451
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3556..3698
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3884..4472)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4646..5503)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5504..5608)
                     /label=AmpR promoter
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
      241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
      301 cgggctttgt tagcagccgg atcctcgagc atatgacgac cttcgatatg gccgctgctg
      361 tgatgatgat gatgatgatg atgatgatgg cccatggtat atctccttct taaagttaaa
      421 caaaattatt tctagagggg aattgttatc cgctcacaat tcccctatag tgagtcgtat
      481 taatttcgcg ggatcgagat ctcgatcctc tacgccggac gcatcgtggc cggcatcacc
      541 ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg
      601 gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg
      661 gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc
      721 aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt
      781 cgagatcccg gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc
      841 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
      901 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
      961 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
     1021 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
     1081 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
     1141 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
     1201 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
     1261 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
     1321 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
     1381 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
     1441 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
     1501 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
     1561 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
     1621 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
     1681 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
     1741 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
     1801 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
     1861 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     1921 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca
     1981 ctcattaggc accgggatct cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     2041 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     2101 aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct
     2161 ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc
     2221 aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg
     2281 gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg
     2341 ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca
     2401 tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg ctcgcggctc
     2461 ttaccagcct aacttcgatc actggaccgc tgatcgtcac ggcgatttat gccgcctcgg
     2521 cgagcacatg gaacgggttg gcatggattg taggcgccgc cctatacctt gtctgcctcc
     2581 ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac ctgaatggaa gccggcggca
     2641 cctcgctaac ggattcacca ctccaagaat tggagccaat caattcttgc ggagaactgt
     2701 gaatgcgcaa accaaccctt ggcagaacat atccatcgcg tccgccatct ccagcagccg
     2761 cacgcggcgc atctcgggca gcgttgggtc ctggccacgg gtgcgcatga tcgtgctcct
     2821 gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc
     2881 gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac
     2941 atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cagcgccctg
     3001 caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac
     3061 atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt cccgccgcat
     3121 ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt catcatcagt
     3181 aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca tgaacagaaa
     3241 tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta acatggcccg
     3301 ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg acgcggatga
     3361 acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc gcagctgcct
     3421 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     3481 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     3541 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg
     3601 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatatg cggtgtgaaa
     3661 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
     3721 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3781 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3841 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3901 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3961 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4021 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4081 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4141 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4201 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4261 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4321 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4381 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4441 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4501 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4561 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4621 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4681 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4741 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4801 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4861 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4921 cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct
     4981 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5041 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5101 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5161 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5221 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     5281 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5341 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5401 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5461 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5521 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5581 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     5641 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5701 gtcttcaaga a
//
