LOCUS       pET-45b(+).        5260 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expressing N-terminally 6xHis-tagged and 
            C-terminally S-tagged proteins.
ACCESSION   .
VERSION     .
KEYWORDS    pET-45b(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5260)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5260)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5260
                     /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(125..169)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     misc_feature    176..248
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(248..262)
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             complement(281..298)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(302..304)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(311..333)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(348..372)
                     /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(373..391)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        704..781
                     /label=lacI promoter
     CDS             782..1861
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1877..1898
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2436..2624
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    2729..2871
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3057..3645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        3726..3818
                     /label=AmpR promoter
     CDS             3819..4676
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      complement(4794..5249)
                     /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 cagcagccta ggttaattaa gctgcgctag
      121 tagacgagtc catgtgctgg cgttcaaatt tcgcagcagc ggtttcttta ccagactcga
      181 gtgcggccgc aagcttgtcg acctgcaggc gcgccgtgta cacgagctcc caattgggat
      241 ccggactctt gtcgtcgtca tcattcgaac cggtacccac gtgatggtgg tggtgatgtg
      301 ccatggtata tctccttctt aaagttaaac aaaattattt ctagagggga attgttatcc
      361 gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc gatctcgatc
      421 ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc
      481 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct
      541 tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc
      601 ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc
      661 ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca tcgaatggcg
      721 caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa
      781 tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt
      841 ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc
      901 ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca
      961 gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt
     1021 cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga
     1081 acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag
     1141 tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg
     1201 cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat
     1261 tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca
     1321 gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg
     1381 ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg
     1441 gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac
     1501 tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc
     1561 cgggctgcgc gttggtgcgg acatctcggt agtgggatac gacgataccg aagacagctc
     1621 atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag
     1681 cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc
     1741 cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg
     1801 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     1861 gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga tctcgaccga
     1921 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactagca
     1981 tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc
     2041 agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga
     2101 cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga
     2161 agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct
     2221 gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct
     2281 ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat
     2341 gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc
     2401 ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc
     2461 ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc
     2521 tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt
     2581 accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     2641 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     2701 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     2761 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     2821 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc
     2881 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2941 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3001 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3061 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3121 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3181 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3241 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3301 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3361 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3421 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3481 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3541 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3601 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3661 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3721 acctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3781 ctgataaatg cttcaatcat gattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3841 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3901 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3961 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4021 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4081 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4141 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4201 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4261 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4321 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4381 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4441 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4501 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4561 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4621 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4681 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4741 aaaggatcta ggtgaagatc ctttttgata atctcatgcc atcgtgccgc cagaaattgt
     4801 aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     4861 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     4921 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     4981 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     5041 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     5101 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     5161 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     5221 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//
