LOCUS       pET-40b(+).        6190 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expressing periplasmic DsbC fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    pET-40b(+).
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6190)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6190)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6190
                     /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(150..173)
                     /label=8xHis
                     /note="8xHis affinity tag"
     misc_feature    174..250
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(252..266)
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             complement(282..326)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             complement(342..359)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(369..386)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(408..1115)
                     /label=DsbC
                     /note="bacterial periplasmic protein disulfide isomerase"
     RBS             complement(1123..1145)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(1160..1184)
                     /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(1185..1203)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1516..1593
                     /label=lacI promoter
     CDS             1594..2673
                     /label=lacI
                     /note="lac repressor"
     protein_bind    2689..2710
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3485..3673
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3778..3920
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4106..4694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             4816..5628
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(5724..6179)
                     /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 tgtttagcag cctaggtatt aatcaattag tggtggtggt ggtggtggtg gtgctcgagt
      181 gcggccgcaa gcttgtcgac ggagctcgaa ttcggatccg atatcgccat ggttgaggag
      241 aagcccgggc tcttgtcgtc gtcatcggta cccagatctg ggctgtccat gtgctggcgt
      301 tcgaatttag cagcagcggt ttctttcata ccaattgcag tactaccgcg tggcaccaga
      361 cccgcggagt gatggtgatg gtgatgacca gaaccactag ttgatccttt accgctggtc
      421 attttttggt gttcgtcgag aaattctttc atctctttcg gcggctggta acccggaaca
      481 agtgtgccat tgctcagcac aactgccgga gtaccgctaa cgccaagctg gacgccaagt
      541 gcgtaatggt cggcaatatc cacgtcgcaa ctggctggtg cgacgctttt acctgccatc
      601 acatcatcaa acgctttgtt tttatctttc gcacaccaga tagctttcat ttctttctct
      661 gcatcgctgt ccagcccctg gcgcgggaaa gcaagataac gcacggtgat ccccagcgcg
      721 ttgtagtctg ccatttgctc atgcagtttg tggcagtaac cacaggtaat atcagtaaac
      781 acggtgatga cgtgtttttc ctgcggcgct ttataaacga tcatctcttt ttcaagcgca
      841 ttcaactgct ttaacagcat cttattggtg acattgaccg gagccgtgcc actaacgtca
      901 tacattggcc cctgaatgat atgtttacca tcatcggtga tgtacaacac gccgctgtta
      961 gtcagaactg tcttcatgcc agctacaggc gcgggctgaa tatcgctgct tttgatgccc
     1021 attttggcta acgtttgttg aattgccgcg tcatcagcct gagcaaagcc tgaaaacgcc
     1081 gctaacaaag taaacaacat aaaacctttc ttcatatgta tatctccttc ttaaagttaa
     1141 acaaaattat ttctagaggg gaattgttat ccgctcacaa ttcccctata gtgagtcgta
     1201 ttaatttcgc gggatcgaga tcgatctcga tcctctacgc cggacgcatc gtggccggca
     1261 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag
     1321 atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc
     1381 ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg
     1441 tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag
     1501 agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag
     1561 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc
     1621 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac
     1681 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc
     1741 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc
     1801 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa
     1861 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg
     1921 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat
     1981 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat
     2041 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg
     2101 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta
     2161 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa
     2221 attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc
     2281 atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg
     2341 gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggacatctcg
     2401 gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc
     2461 aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag
     2521 ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc
     2581 ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     2641 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta
     2701 gctcactcat taggcaccgg gatctcgacc gatgcccttg agagccttca acccagtcag
     2761 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
     2821 catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt
     2881 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct
     2941 cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat
     3001 cgccggcatg gcggccccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct
     3061 aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg
     3121 aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt
     3181 ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat
     3241 ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg
     3301 ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt
     3361 accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat
     3421 cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc
     3481 atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac
     3541 attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga
     3601 atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga
     3661 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     3721 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     3781 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
     3841 gagcagattg tactgagagt gcaccatata tgcggtgtga aataccgcac agatgcgtaa
     3901 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3961 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4021 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4081 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4141 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4201 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4261 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4321 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4381 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4441 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4501 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     4561 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4621 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4681 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4741 aaaactcacg ttaagggatt ttggtcatga acaataaaac tgtctgctta cataaacagt
     4801 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctctag gccgcgatta
     4861 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     4921 tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa
     4981 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     5041 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     5101 ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat
     5161 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     5221 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     5281 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     5341 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     5401 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     5461 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     5521 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     5581 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta agaattaatt
     5641 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5701 atttccccga aaagtgccac ctgaaattgt aaacgttaat attttgttaa aattcgcgtt
     5761 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     5821 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     5881 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     5941 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     6001 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     6061 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     6121 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     6181 ccattcgcca
//
