LOCUS       40924_3846        3259 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pACpET24, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3259)
  AUTHORS   Elce JS, Hegadorn C, Gauthier S, Vince JW, Davies PL.
  TITLE     Recombinant calpain II: improved expression systems and production 
            of a C105A active-site mutant for crystallography
  JOURNAL   Protein Eng. 8 (8), 843-848 (1995)
  PUBMED    8637855
REFERENCE   2  (bases 1 to 3259)
  AUTHORS   Elce JS.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-OCT-1994) John S. Elce, Biochemistry, Queen's 
            University, Kingston, Ontario K7L 3N6, Canada
REFERENCE   3  (bases 1 to 3259)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3259)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Eng."; date: "1995"; volume: "8"; issue: "8"; pages: "843-848"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-OCT-1994) John S. Elce, Biochemistry, Queen's University, 
            Kingston, Ontario K7L 3N6, Canada"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      783..1328
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     CDS             complement(2504..2536)
                     /codon_start=1
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
                     /translation="MASMTGGQQMG"
     RBS             complement(2543..2565)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(2580..2604)
                     /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(2605..2623)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2912..3016
                     /label=AmpR promoter
     CDS             join(3017..3259,1..615)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    3201..3259
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene;
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
ORIGIN
        1 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
       61 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      121 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
      181 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
      241 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
      301 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
      361 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
      421 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
      481 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
      541 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
      601 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
      661 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
      721 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccttaat aagatgatct
      781 tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg
      841 cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt
      901 ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc
      961 aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct
     1021 ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg
     1081 gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg
     1141 tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa
     1201 caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg
     1261 ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc
     1321 tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc
     1381 ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga
     1441 gcgtagcgag tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg
     1501 caccggtgca gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg
     1561 ccaacatagt aagccagtat acactccgct agcgctgagg tctgcctcgt gaagaaggtg
     1621 ttgctgactc ataccaggcc tgaatcgccc catcatccag ccagaaagtg agggagccac
     1681 ggttgatgag agctttgttg taggtggacc agttggtgat tttgaacttt tgctttgcca
     1741 cggaacggtc tgcgttgtcg ggaagatgcg tgatctgatc cttcaactca gcaaaagttc
     1801 gatttattca acaaagccac gttgtgtctc aaaatctctg atgttacatt gcacaagata
     1861 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     1921 ttatgagcca tattcaacgg gaaacgtctt gctcgaggcc gcgattaaat tccaacatgg
     1981 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     2041 tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     2101 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     2161 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     2221 cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     2281 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     2341 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     2401 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     2461 aagaaatgca taagcttgtc gacggagctc gaattcggat ccgacccatt tgctgtccac
     2521 cagtcatgct agccatggta tatctccttc ttaaagttaa acaaaattat ttctagaggg
     2581 gaattgttat ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc gggatcgaga
     2641 tcctagagcg cacgaatgag ggccgacagg aagcaaagct gaaaggaatc aaatttggcc
     2701 gcaggcgtac cgtggacagg aacgtcgtgc tgacgcttca tcagaagggc actggtgcaa
     2761 cggaaattgc tcatcagctc agtattgccc gctccacggt ttataaaatt cttgaagacg
     2821 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     2881 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2941 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3001 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3061 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3121 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3181 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3241 tggcgcggta ttatcccgt
//
