LOCUS       40924_40072        3352 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pSH476, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3352)
  AUTHORS   Hwang S-K., Okita TW.
  TITLE     Protein expression vector pSH476 (N-terminal His-tag)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3352)
  AUTHORS   Hwang S-K., Okita TW.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-APR-2008) Institute of Biological Chemistry, 
            Washington State University, Pullman, WA 99163, USA
REFERENCE   3  (bases 1 to 3352)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3352)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-APR-2008) Institute of Biological Chemistry, Washington State 
            University, Pullman, WA 99163, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3352
                     /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 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)."
     RBS             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             127..144
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             166..876
                     /label=mgfp5
                     /note="GFP with folding enhancement mutations"
     terminator      955..1049
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    1202..1342
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(1340..1440)
                     /direction=LEFT
                     /label=Ori
                     /note="Ori"
     rep_origin      complement(1528..2116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2290..3147)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3148..3252)
                     /label=AmpR promoter
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
      121 ggatcgcatc accatcacca tcacggatcc atggtagatc tgactagtaa aggagaagaa
      181 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa
      241 ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt
      301 atttgcacta ctggaaaact acctgttccg tggccaacac ttgtcactac tttctcttat
      361 ggtgttcaat gcttttcaag atacccagat catatgaagc ggcacgactt cttcaagagc
      421 gccatgcctg agggatacgt gcaggagagg accatcttct tcaaggacga cgggaactac
      481 aagacacgtg ctgaagtcaa gtttgaggga gacaccctcg tcaacaggat cgagcttaag
      541 ggaatcgatt tcaaggagga cggaaacatc ctcggccaca agttggaata caactacaac
      601 tcccacaacg tatacatcat ggccgacaag caaaagaacg gcatcaaagc caacttcaag
      661 acccgccaca acatcgaaga cggcggcgtg caactcgctg atcattatca acaaaatact
      721 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc
      781 ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct
      841 gctgggatta cacatggcat ggatgaacta tacaaagcta gctagatgca ttcgcgaggt
      901 accgagctcg gtaccccggg tcgacctgca gccaagctta attagctgag cttggactcc
      961 tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt
     1021 gccgccgggc gttttttatt ggtgagaatc caagctagag ctgcctcgcg cgtttcggtg
     1081 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     1141 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     1201 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc
     1261 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     1321 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     1381 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     1441 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     1501 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     1561 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     1621 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     1681 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     1741 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1801 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     1861 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1921 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     1981 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2041 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2101 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2161 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2221 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2281 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     2341 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     2401 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     2461 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     2521 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     2581 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     2641 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     2701 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     2761 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     2821 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     2881 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     2941 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3001 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3061 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3121 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     3181 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     3241 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     3301 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc ac
//
