LOCUS       40924_24673        4506 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pHisJM DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4506)
  AUTHORS   Masuda J, Takayama E, Satoh A, Kojima-Aikawa K, Suzuki K, Matsumoto 
            I.
  TITLE     A novel expression vector, designated as pHisJM, for producing 
            recombinant His-fusion proteins
  JOURNAL   Biotechnol. Lett. 26 (20), 1543-1548 (2004)
  PUBMED    15604794
REFERENCE   2  (bases 1 to 4506)
  AUTHORS   Masuda J.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-AUG-2003) Contact:Junko Masuda Ochanomizu University, 
            Graduate School of Humanities and Sciences; 2-1-1 Otsuka, Bunkyo-ku,
            Tokyo 112-8610, Japan
REFERENCE   3  (bases 1 to 4506)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4506)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol.
            Lett."; date: "2004"; volume: "26"; issue: "20"; pages: "1543-1548"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (05-AUG-2003) Contact:Junko Masuda Ochanomizu University, Graduate 
            School of Humanities and Sciences; 2-1-1 Otsuka, Bunkyo-ku, Tokyo 
            112-8610, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        183..211
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    219..235
                     /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)."
     CDS             270..287
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             291..323
                     /codon_start=1
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
                     /translation="MASMTGGQQMG"
     CDS             327..350
                     /codon_start=1
                     /label=Xpress(TM) tag
                     /note="Xpress(TM) epitope tag, including an enterokinase 
                     recognition and cleavage site"
                     /translation="DLYDDDDK"
     terminator      472..519
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        809..913
                     /label=AmpR promoter
     CDS             914..1771
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1945..2533
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        2777..2854
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             2855..3934
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    3950..3971
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3986..4016
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4024..4040
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     CDS             4060..4233
                     /codon_start=1
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
ORIGIN
        1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg gggggttctc atcatcatca tcatcatggt atggctagca
      301 tgactggtgg acagcaaatg ggtcgggatc tgtacgacga tgacgataag gatcgatgga
      361 tccgacctcg agatctgcag ctggtaccat gcatggaatt cgaagcttga tccggctgct
      421 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
      481 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
      541 ggatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
      601 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
      661 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tataattctt gaagacgaaa
      721 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
      781 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
      841 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
      901 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
      961 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1021 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1081 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1141 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     1201 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1261 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1321 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1381 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1441 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1501 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     1561 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     1621 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     1681 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     1741 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     1801 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1861 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1921 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1981 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2041 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     2101 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2161 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2221 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2281 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2341 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2401 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2461 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2521 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2581 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     2641 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     2701 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2761 acaccgcata aattccgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata
     2821 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
     2881 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
     2941 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     3001 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     3061 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     3121 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
     3181 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     3241 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     3301 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
     3361 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     3421 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     3481 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     3541 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
     3601 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
     3661 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
     3721 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
     3781 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
     3841 cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     3901 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     3961 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     4021 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacggattca
     4081 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     4141 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     4201 ccttcccaac agttgcgcag cctgaatggc gaatggcgct ttgcctggtt tccggcacca
     4261 gaagcggtgc cggaaagctg gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc
     4321 ccctcaaact ggcagatgca cggttacgat gcgcccatct acaccaacgt aacctatccc
     4381 attacggtca atccgccgtt tgttcccacg gagaatccga cgggttgtta ctcgctcaca
     4441 tttaatgttg atgaaagctg gctacaggaa ggccagacgc gaattatttt tgatggcgtt
     4501 ggaatt
//
