LOCUS       pET-30a(+).        5422 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expression of N-terminally S-tagged proteins 
            with an enterokinase site. For other reading frames, use pET-30b(+) 
            or pET-30c(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-30a(+).
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5422)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5422)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5422
                     /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..217
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(219..233)
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             complement(249..293)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             complement(300..317)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(327..344)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(345..347)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(355..377)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(392..416)
                     /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(417..435)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        748..825
                     /label=lacI promoter
     CDS             826..1905
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1921..1942
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2717..2905
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3010..3152
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3338..3926)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             4048..4860
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(4956..5411)
                     /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 tccgatatca gccatggcct tgtcgtcgtc gtcggtaccc
      241 agatctgggc tgtccatgtg ctggcgttcg aatttagcag cagcggtttc tttcatacca
      301 gaaccgcgtg gcaccagacc agaagaatga tgatgatgat ggtgcatatg tatatctcct
      361 tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac aattccccta
      421 tagtgagtcg tattaatttc gcgggatcga gatcgatctc gatcctctac gccggacgca
      481 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca
      541 ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta
      601 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc
      661 ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt
      721 cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcaaaac ctttcgcggt
      781 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
      841 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
      901 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
      961 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     1021 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     1081 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     1141 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     1201 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     1261 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     1321 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     1381 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     1441 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     1501 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     1561 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     1621 gcggacatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     1681 ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     1741 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     1801 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     1861 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     1921 taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct tgagagcctt
     1981 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     2041 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg
     2101 cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat
     2161 cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa
     2221 gcaggccatt atcgccggca tggcggcccc acgggtgcgc atgatcgtgc tcctgtcgtt
     2281 gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg
     2341 cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat
     2401 ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat
     2461 tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt
     2521 attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac
     2581 cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg
     2641 tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc
     2701 ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat
     2761 cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc
     2821 agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg
     2881 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     2941 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     3001 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac
     3061 tatgcggcat cagagcagat tgtactgaga gtgcaccata tatgcggtgt gaaataccgc
     3121 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     3181 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3241 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3301 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3361 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3421 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3481 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3541 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3601 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3661 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3721 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     3781 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3841 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3901 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3961 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gaacaataaa actgtctgct
     4021 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcttgctct
     4081 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     4141 aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag
     4201 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     4261 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     4321 gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca ggtattagaa
     4381 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     4441 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     4501 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     4561 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     4621 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     4681 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     4741 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     4801 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     4861 taagaattaa ttcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4921 ggttccgcgc acatttcccc gaaaagtgcc acctgaaatt gtaaacgtta atattttgtt
     4981 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     5041 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     5101 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     5161 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     5221 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     5281 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     5341 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     5401 acagggcgcg tcccattcgc ca
//
