LOCUS       pET-22b(+).        5493 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector that encodes a signal sequence for inducible 
            expression of proteins in the periplasm.
ACCESSION   .
VERSION     .
KEYWORDS    pET-22b(+).
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5493)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5493)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5493
                     /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"
     misc_feature    158..225
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     sig_peptide     complement(224..289)
                     /label=pelB signal sequence
                     /note="leader peptide for secretion"
     RBS             complement(297..319)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(334..358)
                     /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(359..377)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        686..763
                     /label=lacI promoter
     CDS             764..1843
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1859..1880
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2655..2843
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    2948..3090
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3276..3864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4038..4895)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4896..5000)
                     /label=AmpR promoter
     rep_origin      complement(5027..5482)
                     /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 gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccgaattaa ttccgatatc catggccatc gccggctggg
      241 cagcgaggag cagcagacca gcagcagcgg tcggcagcag gtatttcata tgtatatctc
      301 cttcttaaag ttaaacaaaa ttatttctag aggggaattg ttatccgctc acaattcccc
      361 tatagtgagt cgtattaatt tcgcgggatc gagatctcga tcctctacgc cggacgcatc
      421 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
      481 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
      541 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt
      601 gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg
      661 cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat
      721 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
      781 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
      841 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
      901 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
      961 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
     1021 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
     1081 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
     1141 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
     1201 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
     1261 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
     1321 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
     1381 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
     1441 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
     1501 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
     1561 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     1621 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     1681 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
     1741 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     1801 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     1861 atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg agagccttca
     1921 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
     1981 tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg
     2041 aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct
     2101 tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc
     2161 aggccattat cgccggcatg gcggccccac gggtgcgcat gatcgtgctc ctgtcgttga
     2221 ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg
     2281 agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg
     2341 tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta
     2401 tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat
     2461 taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg
     2521 ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta
     2581 tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt
     2641 acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca
     2701 gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag
     2761 acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt
     2821 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     2881 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     2941 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
     3001 tgcggcatca gagcagattg tactgagagt gcaccatata tgcggtgtga aataccgcac
     3061 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
     3121 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3181 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3241 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3301 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3361 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3421 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3481 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3541 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3601 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3661 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     3721 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3781 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3841 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3901 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3961 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4021 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4081 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4141 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4201 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4261 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4321 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
     4381 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4441 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4501 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4561 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4621 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4681 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4741 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4801 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4861 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4921 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4981 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaaat tgtaaacgtt
     5041 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     5101 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     5161 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     5221 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     5281 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     5341 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     5401 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     5461 aatgcgccgc tacagggcgc gtcccattcg cca
//
