LOCUS       sequence-445437-        5058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG induces expression of C. elegans protein MDH-2 with a 
            N-terminal His tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5058)
  AUTHORS   Katherine Walstrom Lab
  TITLE     Walstrom Lab Plasmids 2024
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5058)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: MDH2_EC
FEATURES             Location/Qualifiers
     source          1..5058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             332..349
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             353..385
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(389..397,398..412)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(1458..1475)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1628..1714
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1806..1833
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1853..1944
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1945..2013,2014..2805)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2163..2182)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2976..3564
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3465..3484
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3718..3735
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3750..3890
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3816..3835)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3976..3998
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4154..5032)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctaacaggag gaattaacca tggggggttc tcatcatcat catcatcatg gtatggctag
      361 catgactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatcgatg
      421 gggatccgag ctcgagatct gcagcagcca agctccaaag gtcgctcttc ttggagctgc
      481 cggaggaatt ggacagccat tgggtctcct tctcaagcag gatccacttg ttgctcacct
      541 cgccctctac gacgtcgtca atactccagg agttgccgcc gatctttcgc acattgattc
      601 taatgccaag gttaccgctc acactggccc aaaggaactt tacgccgccg ttgagaacgc
      661 tgatgttatc gttattccag ccggagttcc acgtaaacca ggaatgactc gtgacgatct
      721 cttcaacacc aacgcaggaa tcgttcgcga tttagctgcc gttatcgcca aagcttcacc
      781 aaaggctctc atcgcaatta tcaccaaccc agtcaattct actgtcccaa tcgcttccga
      841 agttctcaag aaggctggtg tctatgaccc aaaacgtgtc ttcggagtga caaccctcga
      901 cgtcgtcaga tctcaagcat tcgtttccga gcttaaggga catgacgcct ccaagaccgt
      961 tgttccagtt gtcggaggac atgctggaat caccatcatt ccacttctct cccaagtcaa
     1021 accatccacc aaattctcgg aggaagaaat ctccaagttg accccaagaa tccaggacgc
     1081 cggaactgaa gtcgtcaacg ccaaggctgg agccggatcc gctactcttt caatggctct
     1141 tgctggagcc cgtttcgcca atgctcttgt tcgtggaatt aagggagaaa agaacgtcca
     1201 atgcgcatat gttgcttccg atgccgtcaa gggagtcgaa tacttctcta ccccagttga
     1261 gctcggacca aatggagttg agaagattct tggagttgga aaggtttctg cttatgagca
     1321 gaagcttatc gacgcatccg ttccagaact taacaagaac attgccaagg gagtcgcttt
     1381 cgttaaggga aactaaagct ggtaccatat gggaattcga agcttggctg ttttggcgga
     1441 tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa
     1501 cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa
     1561 gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc
     1621 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     1681 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
     1741 gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat
     1801 taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc tttttgttta
     1861 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     1921 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     1981 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2041 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2101 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2161 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     2221 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     2281 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2341 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2401 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2461 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2521 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2581 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2641 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2701 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2761 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     2821 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     2881 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     2941 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3001 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3061 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3121 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3181 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3241 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3301 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3361 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3421 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3481 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3541 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3601 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3661 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     3721 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     3781 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     3841 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     3901 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     3961 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     4021 gcgcgaggca gcagatcaat tcgcgcgcga aggcgaagcg gcatgcataa tgtgcctgtc
     4081 aaatggacga agcagggatt ctgcaaaccc tatgctactc cgtcaagccg tcaattgtct
     4141 gattcgttac caattatgac aacttgacgg ctacatcatt cactttttct tcacaaccgg
     4201 cacggaactc gctcgggctg gccccggtgc attttttaaa tacccgcgag aaatagagtt
     4261 gatcgtcaaa accaacattg cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa
     4321 gcagcttcgc ctggctgata cgttggtcct cgcgccagct taagacgcta atccctaact
     4381 gctggcggaa aagatgtgac agacgcgacg gcgacaagca aacatgctgt gcgacgctgg
     4441 cgatatcaaa attgctgtct gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc
     4501 gattatccat cggtggatgg agcgactcgt taatcgcttc catgcgccgc agtaacaatt
     4561 gctcaagcag atttatcgcc agcagctccg aatagcgccc ttccccttgc ccggcgttaa
     4621 tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc
     4681 ccgtattggc aaatattgac ggccagttaa gccattcatg ccagtaggcg cgcggacgaa
     4741 agtaaaccca ctggtgatac cattcgcgag cctccggatg acgaccgtag tgatgaatct
     4801 ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac aaattctcgt ccctgatttt
     4861 tcaccacccc ctgaccgcga atggtgagat tgagaatata acctttcatt cccagcggtc
     4921 ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc gccaccagat
     4981 gggcattaaa cgagtatccc ggcagcaggg gatcattttg cgcttcagcc atacttttca
     5041 tactcccgcc attcagag
//
