LOCUS       TAGZyme_pQE-2.        4758 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector with a lacIq repressor gene for bacterial expression of 
            6xHis-tagged proteins that can be processed by TAGZyme.
ACCESSION   .
VERSION     .
KEYWORDS    TAGZyme pQE-2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4758)
  AUTHORS   Qiagen
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4758)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     For TAGZyme cleavage, a glutamine should be inserted at an uneven 
            amino acid position, followed by the first amino acid of the 
            expressed protein.
FEATURES             Location/Qualifiers
     source          1..4758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..54
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
     protein_bind    62..78
                     /label=lac repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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)."
     RBS             101..106
                     /note="ribosome binding site"
     CDS             115..117
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             121..141
                     /label=7xHis
                     /note="6xHis affinity tag"
     misc_feature    139..199
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     misc_feature    199..209
                     /label=stop codons
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     terminator      215..309
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             353..1009
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     terminator      1077..1163
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     protein_bind    complement(1222..1243)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(1259..2338)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(2339..2416)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     misc_feature    2608..2748
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2934..3522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3696..4553)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4554..4658)
                     /label=AmpR promoter
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaaa
      121 catcaccatc accatcacca tatgcatgcg ggagctcatg cggccgcggg tacccacgtg
      181 tcgacctgca gccaagctta attagctgag cttggactcc tgttgataga tccagtaatg
      241 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
      301 ggtgagaatc caagctagct tggcgagatt ttcaggagct aaggaagcta aaatggagaa
      361 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
      421 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
      481 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
      541 tgcccgcctg atgaatgctc atccggaatt tcgtatggca atgaaagacg gtgagctggt
      601 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
      661 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga
      721 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
      781 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
      841 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt
      901 gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag
      961 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aattttttta
     1021 aggcagttat tggtgccctt aaacgcctgg ggtaatgact ctctagcttg aggcatcaaa
     1081 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     1141 acgctctcct gagtaggaca aatccgccct ctagattacg tgcagtcgat gataagctgt
     1201 caaacatgag aattgtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     1261 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     1321 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     1381 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     1441 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     1501 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg
     1561 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     1621 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     1681 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     1741 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     1801 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     1861 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     1921 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     1981 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     2041 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     2101 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     2161 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     2221 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     2281 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     2341 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     2401 accattcgat ggtgtcggaa tttcgggcag cgttgggtcc tggccacggg tgcgcatgat
     2461 ctagagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     2521 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     2581 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     2641 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccacatg
     2701 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
     2761 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2821 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2881 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2941 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3001 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3061 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3121 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3181 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3241 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3301 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3361 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3421 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3481 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3541 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3601 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3661 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3721 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3781 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3841 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3901 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3961 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4021 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4081 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4141 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4201 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4261 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4321 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4381 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4441 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4501 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4561 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4621 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4681 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     4741 aggccctttc gtcttcac
//
