LOCUS       pET-15b.        5708 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for inducible expression of N-terminally 
            6xHis-tagged proteins.
ACCESSION   .
VERSION     .
KEYWORDS    pET-15b
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5708)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5708)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5708
                     /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..353)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(363..380)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(390..392)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(399..421)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(436..460)
                     /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(461..479)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        788..865
                     /label=lacI promoter
     CDS             866..1945
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1961..1982
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3260..3448
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3553..3695
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3881..4469)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4643..5500)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5501..5605)
                     /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 atatggctgc cgcgcggcac caggccgctg
      361 ctgtgatgat gatgatgatg gctgctgccc atggtatatc tccttcttaa agttaaacaa
      421 aattatttct agaggggaat tgttatccgc tcacaattcc cctatagtga gtcgtattaa
      481 tttcgcggga tcgagatctc gatcctctac gccggacgca tcgtggccgg catcaccggc
      541 gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct
      601 cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc
      661 gggggactgt tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac
      721 ggcctcaacc tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga
      781 gatcccggac accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga
      841 agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta
      901 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
      961 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt
     1021 ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc
     1081 cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc
     1141 cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca
     1201 caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga
     1261 tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga
     1321 ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga
     1381 gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt
     1441 ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc
     1501 gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat
     1561 gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg
     1621 cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg
     1681 atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga
     1741 ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc
     1801 ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc
     1861 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     1921 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc
     1981 attaggcacc gggatctcga ccgatgccct tgagagcctt caacccagtc agctccttcc
     2041 ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac
     2101 tcgtaggaca ggtgccggca gcgctctggg tcattttcgg cgaggaccgc tttcgctgga
     2161 gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag
     2221 ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt atcgccggca
     2281 tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct
     2341 tccccattat gattcttctc gcttccggcg gcatcgggat gcccgcgttg caggccatgc
     2401 tgtccaggca ggtagatgac gaccatcagg gacagcttca aggatcgctc gcggctctta
     2461 ccagcctaac ttcgatcact ggaccgctga tcgtcacggc gatttatgcc gcctcggcga
     2521 gcacatggaa cgggttggca tggattgtag gcgccgccct ataccttgtc tgcctccccg
     2581 cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg aatggaagcc ggcggcacct
     2641 cgctaacgga ttcaccactc caagaattgg agccaatcaa ttcttgcgga gaactgtgaa
     2701 tgcgcaaacc aacccttggc agaacatatc catcgcgtcc gccatctcca gcagccgcac
     2761 gcggcgcatc tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg tgctcctgtc
     2821 gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat
     2881 acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg
     2941 aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac
     3001 cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc
     3061 tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca
     3121 taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac
     3181 ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc
     3241 cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt
     3301 tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca
     3361 ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc
     3421 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     3481 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     3541 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
     3601 aactatgcgg catcagagca gattgtactg agagtgcacc atatatgcgg tgtgaaatac
     3661 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     3721 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3781 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3841 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3901 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3961 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4021 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4081 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4141 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4201 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4261 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4321 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4381 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4441 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4501 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4561 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4621 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4681 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4741 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4801 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4861 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4921 cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt
     4981 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5041 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5101 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5161 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5221 gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata
     5281 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5341 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5401 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5461 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5521 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5581 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     5641 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5701 ttcaagaa
//
