LOCUS       sequence-444697-        5052 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 5052)
  AUTHORS   Katherine Walstrom Lab
  TITLE     Walstrom Lab Plasmids 2024
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5052)
  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            "Unpublished"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..5052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(324..346)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    432..572
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     487..506
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(587..604)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(758..1346)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(838..857)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1517..2308,2309..2377))
                     /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     2140..2159
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2378..2469)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(2488..2515)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2607..2693)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2846..2863
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(join(3905..3919,3920..3928))
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(3932..3964)
                     /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             complement(3968..3985)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(4031..4315)
                     /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     complement(4089..4108)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             join(4342..5052,1..168)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 cttttgagca ccacccggat gcctatcgcc accgtcggtc gcaatgttgg ttttgacgat
       61 caactctatt tctcgcgggt atttaaaaaa tgcaccgggg ccagcccgag cgagttccgt
      121 gccggttgtg aagaaaaagt gaatgatgta gccgtcaagt tgtcataatt ggtaacgaat
      181 cagacaattg acggcttgac ggagtagcat agggtttgca gaatccctgc ttcgtccatt
      241 tgacaggcac attatgcatg ccgcttcgcc ttcgcgcgcg aattgatctg ctgcctcgcg
      301 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
      361 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
      421 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
      481 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
      541 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
      601 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
      661 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
      721 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
      781 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
      841 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
      901 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
      961 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1021 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1081 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1141 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     1201 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1261 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1321 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1381 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     1441 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1501 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1561 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     1621 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     1681 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     1741 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     1801 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     1861 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     1921 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     1981 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     2041 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     2101 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
     2161 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     2221 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     2281 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     2341 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     2401 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     2461 ataaacaaaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
     2521 atttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
     2581 gcaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
     2641 caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctg
     2701 gcagttccct actctcgcat ggggagaccc cacactacca tcggcgctac ggcgtttcac
     2761 ttctgagttc ggcatggggt caggtgggac caccgcgcta ctgccgccag gcaaattctg
     2821 ttttatcaga ccgcttctgc gttctgattt aatctgtatc aggctgaaaa tcttctctca
     2881 tccgccaaaa cagccaagct tcgaattccc atatggtacc ttagtttccc ttaacgaaag
     2941 cgactccctt ggcaatgttc ttgttaagtt ctggaacgga tgcgtcgata agcttctgct
     3001 cataagcaga aacctttcca actccaagaa tcttctcaac tccatttggt ccgagctcaa
     3061 ctggggtaga gaagtattcg actcccttga cggcatcgga agcaacatat gcgcattgga
     3121 cgttcttttc tcccttaatt ccacgaacaa gagcattggc gaaacgggct ccagcaagag
     3181 ccattgaaag agtagcggat ccggctccag ccttggcgtt gacgacttca gttccggcgt
     3241 cctggattct tggggtcaac ttggagattt cttcctccga gaatttggtg gatggtttga
     3301 cttgggagag aagtggaatg atggtgattc cagcatgtcc tccgacaact ggaacaacgg
     3361 tcttggaggc gtcatgtccc ttaagctcgg aaacgaatgc ttgagatctg acgacgtcga
     3421 gggttgtcac tccgaagaca cgttttgggt catagacacc agccttcttg agaacttcgg
     3481 aagcgattgg gacagtagaa ttgactgggt tggtgataat tgcgatgaga gcctttggtg
     3541 aagctttggc gataacggca gctaaatcgc gaacgattcc tgcgttggtg ttgaagagat
     3601 cgtcacgagt cattcctggt ttacgtggaa ctccggctgg aataacgata acatcagcgt
     3661 tctcaacggc ggcgtaaagt tcctttgggc cagtgtgagc ggtaaccttg gcattagaat
     3721 caatgtgcga aagatcggcg gcaactcctg gagtattgac gacgtcgtag agggcgaggt
     3781 gagcaacaag tggatcctgc ttgagaagga gacccaatgg ctgtccaatt cctccggcag
     3841 ctccaagaag agcgaccttt ggagcttggc tgctgcagat ctcgagctcg gatccccatc
     3901 gatccttatc gtcatcgtcg tacagatccc gacccatttg ctgtccacca gtcatgctag
     3961 ccataccatg atgatgatga tgatgagaac cccccatggt taattcctcc tgttagccca
     4021 aaaaacgggt atggagaaac agtagagagt tgcgataaaa agcgtcaggt aggatccgct
     4081 aatcttatgg ataaaaatgc tatggcatag caaagtgtga cgccgtgcaa ataatcaatg
     4141 tggacttttc tgccgtgatt atagacactt ttgttacgcg tttttgtcat ggctttggtc
     4201 ccgctttgtt acagaatgct tttaataagc ggggttaccg gtttggttag cgagaagagc
     4261 cagtaaaaga cgcagtgacg gcaatgtctg atgcaatatg gacaattggt ttcttctctg
     4321 aatggcggga gtatgaaaag tatggctgaa gcgcaaaatg atcccctgct gccgggatac
     4381 tcgtttaatg cccatctggt ggcgggttta acgccgattg aggccaacgg ttatctcgat
     4441 ttttttatcg accgaccgct gggaatgaaa ggttatattc tcaatctcac cattcgcggt
     4501 cagggggtgg tgaaaaatca gggacgagaa tttgtttgcc gaccgggtga tattttgctg
     4561 ttcccgccag gagagattca tcactacggt cgtcatccgg aggctcgcga atggtatcac
     4621 cagtgggttt actttcgtcc gcgcgcctac tggcatgaat ggcttaactg gccgtcaata
     4681 tttgccaata cggggttctt tcgcccggat gaagcgcacc agccgcattt cagcgacctg
     4741 tttgggcaaa tcattaacgc cgggcaaggg gaagggcgct attcggagct gctggcgata
     4801 aatctgcttg agcaattgtt actgcggcgc atggaagcga ttaacgagtc gctccatcca
     4861 ccgatggata atcgggtacg cgaggcttgt cagtacatca gcgatcacct ggcagacagc
     4921 aattttgata tcgccagcgt cgcacagcat gtttgcttgt cgccgtcgcg tctgtcacat
     4981 cttttccgcc agcagttagg gattagcgtc ttaagctggc gcgaggacca acgtatcagc
     5041 caggcgaagc tg
//
