LOCUS       sequence-411058-        4365 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of R196M wgMDH with IPTG induction - Active 
            site mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4365)
  AUTHORS   Cox B, Chit MM, Weaver T, Gietl C, Bailey J, Bell E, Banaszak L
  TITLE     Organelle and translocatable forms of glyoxysomal malate 
            dehydrogenase. The effect of the N-terminal presequence.
  JOURNAL   FEBS J. 2005 Feb;272(3):643-54. doi: 
            10.1111/j.1742-4658.2004.04475.x.
  PUBMED    15670147
REFERENCE   2  (bases 1 to 4365)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1111/j.1742-4658.2004.04475"; journalName: "FEBS J"; date: 
            "2005-02"; volume: "272"; issue: "3"; pages: "643-54"
COMMENT     Sequence Label: pQE60 R196M wgMDH
FEATURES             Location/Qualifiers
     source          1..4365
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    30..46
                     /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)."
     protein_bind    49..65
                     /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             84..95
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             553..564
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1068..1085
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1108..1202
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1312..1331)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1974..2060
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2107..2129)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2215..2355
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2270..2289
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2370..2387)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2541..3129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2621..2640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3300..4091,4092..4160))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3923..3942
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4161..4265)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4333..4351
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaatttt gtgagcggat
       61 aacaatttca cacagaattc attaaagagg agaaattaac tatggctaaa ggcggagctc
      121 ccgggttcaa agtcgcaata cttggcgctg ccggtggcat tggccagccc cttgcgatgt
      181 tgatgaagat gaatcctctg gtttctgttc tacatctata tgatgtagtc aatgcccctg
      241 gtgtcaccgc tgatattagc cacatggaca cgggtgctgt ggtgcgtgga ttcttggggc
      301 agcagcagct ggaggctgcg cttactggca tggatcttat tatagtccct gcaggtgttc
      361 ctcgaaaacc aggaatgacg agggatgatc tgttcaaaat aaacgcagga attgtcaaga
      421 ctctgtgtga agggattgca aagtgttgtc caagagccat tgtcaacctg atcagtaatc
      481 ctgtgaactc caccgtgccc atcgcagctg aagttttcaa gaaggctgga acttatgatc
      541 caaagcgact tctgggagtt acaatgctcg acgtagtcat ggccaatacc tttgtggcag
      601 aagtattggg tcttgatcct cgggatgttg atgttccagt tgttggcggt catgctggtg
      661 taaccatttt gccccttcta tctcaggtga agcctccaag ttctttcaca caagaagaga
      721 ttagttacct gactgatagg attcaaaatg gtggaacaga agttgtcgag gccaaagcag
      781 gagctggttc agcaactctc tcaatggctt atgctgccgt taagtttgca gatgcatgcc
      841 tcaggggctt aagaggagat gctggtgtca ttgaatgcgc gtttgtgtct tctcaggtga
      901 ctgaacttcc attctttgca tcaaaagtac gacttggtcg caatggtatc gaagaagtat
      961 actcccttgg cccgctaaat gagtatgaga ggattggatt ggagaaagcg aagaaagagt
     1021 tggcaggaag cattgagaag ggagtttcct tcatcagaag cagatctcat caccatcacc
     1081 atcactaagc ttaattagct gagcttggac tcctgttgat agatccagta atgacctcag
     1141 aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga
     1201 atccaagcta gcttggcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     1261 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     1321 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     1381 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     1441 ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct ggtgatatgg
     1501 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     1561 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     1621 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     1681 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     1741 ttcttcgccc ccgttttcac catgcatggg caaatattat acgcaaggcg acaaggtgct
     1801 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     1861 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg
     1921 cagttattgg tgcccttaaa cgcctggggt aatgactctc tagcttgagg catcaaataa
     1981 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     2041 ctctcctgag taggacaaat ccgccctcta gagctgcctc gcgcgtttcg gtgatgacgg
     2101 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     2161 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     2221 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
     2281 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     2341 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2401 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2461 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2521 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2581 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2641 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2701 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2761 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2821 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2881 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2941 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     3001 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3061 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3121 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3181 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3241 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3301 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3361 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3421 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3481 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3541 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3601 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3661 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3721 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3781 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3841 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3901 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3961 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4021 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4081 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4141 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4201 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4261 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     4321 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcac
//
