LOCUS       sequence-411051-        5101 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of a fusion of watermelon glyoxysomal malate 
            dehydrogenase (MDH) and GFP with His tag; Use in CURES (integration 
            of research into undergraduate teaching labs)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5101)
  TITLE     Expression of His tagged recombinant malate dehydrogenase (MDH)
REFERENCE   2  (bases 1 to 5101)
  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"
COMMENT     Sequence Label: pQE60 MGH
FEATURES             Location/Qualifiers
     source          1..5101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(14..28,29..34,35..51,52..58)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    34..50
                     /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    66..82
                     /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             101..112
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             570..581
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(1118..1146)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1741..1761
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1808..1825
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1848..1942
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2052..2071)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2714..2800
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2847..2869)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2955..3095
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3010..3029
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3110..3127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3281..3869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4040..4831,4832..4900))
                     /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     4663..4682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4901..5005)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5073..5091
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcacctcgag aaatcataaa aaatttattt gctttgtgag cggataacaa ttataataga
       61 ttcaattgtg agcggataac aatttcacac agaattcatt aaagaggaga aattaactat
      121 ggctaaaggc ggagctcccg ggttcaaagt cgcaatactt ggcgctgccg gtggcattgg
      181 ccagcccctt gcgatgttga tgaagatgaa tcctctggtt tctgttctac atctatatga
      241 tgtagtcaat gcccctggtg tcaccgctga tattagccac atggacacgg gtgctgtggt
      301 gcgtggattc ttggggcagc agcagctgga ggctgcgctt actggcatgg atcttattat
      361 agtccctgca ggtgttcctc gaaaaccagg aatgacgagg gatgatctgt tcaaaataaa
      421 cgcaggaatt gtcaagactc tgtgtgaagg gattgcaaag tgttgtccaa gagccattgt
      481 caacctgatc agtaatcctg tgaactccac cgtgcccatc gcagctgaag ttttcaagaa
      541 ggctggaact tatgatccaa agcgacttct gggagttaca atgctcgacg tagtcagagc
      601 caataccttt gtggcagaag tattgggtct tgatcctcgg gatgttgatg ttccagttgt
      661 tggcggtcat gctggtgtaa ccattttgcc ccttctatct caggtgaagc ctccaagttc
      721 tttcacacaa gaagagatta gttacctgac tgataggatt caaaatggtg gaacagaagt
      781 tgtcgaggcc aaagcaggag ctggttcagc aactctctca atggcttatg ctgccgttaa
      841 gtttgcagat gcatgcctca ggggcttaag aggagatgct ggtgtcattg aatgcgcgtt
      901 tgtgtcttct caggtgactg aacttccatt ctttgcatca aaagtacgac ttggtcgcaa
      961 tggtatcgaa gaagtatact cccttggccc gctaaatgag tatgagagga ttggattgga
     1021 gaaagcgaag aaagagttgg caggaagcat tgagaaggga gtttccttca tcagaagcag
     1081 atctatggct agcaaaggag aagaactctt cactggagtt gtcccaattc ttgttgaatt
     1141 agatggtgat gttaacggcc acaagttctc tgtcagtgga gagggtgaag gtgatgcaac
     1201 atacggaaaa cttaccctga agttcatctg cactactggc aaactgcctg ttccatggcc
     1261 aacactagtc actactctgt gctatggtgt tcaatgcttt tcaagatacc cggatcatat
     1321 gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg aaaggaccat
     1381 cttcttcaaa gatgacggca actacaagac acgtgctgaa gtcaagtttg aaggtgatac
     1441 ccttgttaat agaatcgagt taaaaggtat tgacttcaag gaagatggca acattctggg
     1501 acacaaattg gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa
     1561 gaatggaatc aaagtgaact tcaagacccg ccacaacatt gaagatggaa gcgttcaact
     1621 agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa
     1681 ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat
     1741 ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg aactgtacaa
     1801 cggatctcat caccatcacc atcactaagc ttaattagct gagcttggac tcctgttgat
     1861 agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg
     1921 ggcgtttttt attggtgaga atccaagcta gcttggcgag attttcagga gctaaggaag
     1981 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     2041 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     2101 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     2161 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag
     2221 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     2281 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     2341 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     2401 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     2461 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgcatggg caaatattat
     2521 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     2581 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     2641 ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggggt aatgactctc
     2701 tagcttgagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     2761 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccctcta gagctgcctc
     2821 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     2881 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     2941 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     3001 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     3061 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     3121 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3181 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3241 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3301 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3361 aaagatacca ggcgttcccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3421 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3481 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3541 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3601 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3661 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3721 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3781 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3841 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3901 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3961 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4021 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4081 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4141 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4201 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4261 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4321 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4381 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4441 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4501 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4561 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4621 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4681 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4741 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4801 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4861 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4921 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4981 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     5041 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5101 t
//
