LOCUS       sequence-267850-        6563 bp    DNA     circular SYN 14-APR-2025
DEFINITION  A recombinant plasmid expressing mdh (malate dehydrogenase) gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6563)
  AUTHORS   Guarnieri MT, Chou YC, Salvachua D, Mohagheghi A, St John PC, 
            Peterson DJ, Bomble YJ, Beckham GT
  TITLE     Metabolic Engineering of Actinobacillus succinogenes Provides 
            Insights into Succinic Acid Biosynthesis.
  JOURNAL   Appl Environ Microbiol. 2017 Aug 17;83(17). pii: AEM.00996-17. doi: 
            10.1128/AEM.00996-17. Print 2017 Sep 1.
  PUBMED    28625987
REFERENCE   2  (bases 1 to 6563)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl 
            Environ Microbiol."; date: "2017-08-17"; volume: "83(17). pii: 
            AEM.00996-17. doi"; pages: " 10.1128/AEM.00996-17. Print 2017 Sep 1"
COMMENT     Sequence Label: pMDH
FEATURES             Location/Qualifiers
     source          1..6563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     214..231
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1686..1703)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1686..1702)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1695..1717)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4453..4471)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4539..4643
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4644..4712,4713..5504)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4862..4881)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    6115..6136
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6151..6156,6157..6174,6175..6181)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6189..6205
                     /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)."
     primer_bind     6194..6216
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6213..6229
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6213..6229
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6524..6543
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
       61 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      121 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      181 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaacgcgaa
      241 gagcgcctga tgcggaagct ttcgataaat tgaaaatgca gcaatagagg aaacacggtt
      301 tgtttgaggc ttcccattaa tcaaacggcg gattttaact gattgatacc aaacaaaaag
      361 gattaggttt gcataaaaac ctaatccttt aatcggtttt gttaaattat tttggtgaat
      421 tattcgccgg taacaaattt cacaccgtta tcaatgtcag tttgcaaggt cggtaacatc
      481 gcttcaagcg cttgttgctc aaatgcgctt aatgtaccta acggcagaat ttcttctacg
      541 ccgtttttac ccaaacgaac cggttgtgca aagaaacggg cgaatttacc gtcgccttca
      601 atataggcgc attccgtaat cacttcgccg tttaatgcgc gagccatgga aaccacaaaa
      661 cgcattgccg catacgccat ggaaagtgta gcggaacccg caccggcttt agcttctacc
      721 acttcggtgc cggcattttg gatgcggtga gttaaatctt ttaattcctg ctcggtaaat
      781 tccacatgcg gaatttgtga caacaaaggt aaaatcgtca cacctgaatg tccgccgata
      841 actgaaatat cgttacggtt gatttcaata tttttcgctt gcacgataaa tttttcggaa
      901 cggatggtat ccagcgtagt aataccgaat aatttccgtt tatcgtatac gcccgctttt
      961 ttcagcactt ccgccgcaat cggaataatc gagttcaccg gattagtgat aatgcaaaca
     1021 caagcctgcg ggcaaacttg cgcaactttt tccactaaat tctggataat accggcgtta
     1081 acattaaaca aatccgcacg agtcataccc ggcttacgcg ccacacccgc acagattaaa
     1141 acgatatccg ccccttttaa tgcctcggac ggatcatcgc cgccgaaacc ttccacttca
     1201 accgaagtcg gaatatggct gatgtctttc gccacaccgg gggtgaccgg cgcaacatcg
     1261 tataagctta aatcgctttc tgccggaaga tgtaatttta acaacaatga aagaggttga
     1321 ccgataccgc cgctggcgcc taataaggtt actttcatga gatactcctc gataatatta
     1381 ataacttgaa ctgggtagat tttacgaaga aaaaaaatga aactcaatta aatattcatc
     1441 tatttttacg atctaaaaca agtttttatc atttattttt cgaaaacgct caaatcacaa
     1501 taaacttaaa tttttattga ttaattttga atttttatgc agaatttatg caatatttcg
     1561 acttaaaaac tgatatattt ctatttatga ccgacaataa aacagataac ctattacagg
     1621 tttttaaagc gttactcgcg caggaacgct tcggttcggg atccccgggt accgagctcg
     1681 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     1741 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     1801 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgccagat gaagcagagc
     1861 gcgccttcgc tgtcctattc aatgtattgg ggatcgaagc ccccgaccgc gaacgccttg
     1921 ccttctatct gcgattggac cctctgactt ggggttgatg ttcatgccgc ctgtttttcc
     1981 tgctcattgg cacgtttcgc aacctgttct cattgcggac accttttcca gcctcgtttg
     2041 gaaagtttca ttgccagtaa taccaatgct ttagaaagaa aaagggatcg aacttttgac
     2101 attcgatccc tttttctgta atctgtttcg tgcgttcttt gctaagatac agaccctaga
     2161 caagtcatat cttagcaaag ggtagctagt aatgcaagag attgcgaagc gtccctacta
     2221 ccaaaaaacc attcaacgac gtaaacagac aaacgcaaac cttaaattag acggtcttca
     2281 gctcggactt cggaagaata aacaggcgta gaagtgataa cgttcttaat acgaaaatta
     2341 agctctgtct ccgtttcgtg ctacggttag aaaggcgaaa gccccaagaa atacaagcac
     2401 acctgataag cgagatttaa ggataacagc gaaattcaat agggtctgaa tttccaaact
     2461 aggttaaatg ccacgacgtt ttattgttgc cccattcaag caacatttga gaaccgaata
     2521 gaaatctttt agtaaaaagc gttctttttt gggtcagcgg ttaatgtgga cggtttaacg
     2581 gtttttcccc tgcgggtcgt attggaaagc cattgaaaag ctgatggata actctgcgag
     2641 ttacccacga gctttccaac agctttccaa cactaaaaac ctaccgccca caataaccac
     2701 ttccctaata ataaaatttt tttattttta ttttggttca aaggctcacg atgttcgcct
     2761 aataaaacga agtcgcctat cggctccgct gatttttata tatcactctc ggggcttttg
     2821 gtgtactatt gtcttttgta atagcaagga cacaaaaagg gtactcttcg agtttccttt
     2881 ttgaccttgc aaaagggctt tgcccccttg acccccgacc gctttcagcg gtcaaaatag
     2941 aagaacggac accattatga aacgtgagaa agagataaaa atcaggctca ccgaaaacga
     3001 gtatcaagcc ttgttagaga gaaaaacgaa agcaaggctt gcggagtggg ttcgggaagt
     3061 tgccctggaa cagcaaccta agcgacagcc gaaagtaatc gaccctgcgt tactgttcga
     3121 gctgaaccgc ataggcgtaa acctgaacca aatcgcccga caatgcaaca gtcaaaagcc
     3181 gagcattgac cttgttagcg tgttggcgac cttgcgagaa attgaaaaaa atctcaaaaa
     3241 attgcgagaa ttgagcctat gatcgttaaa ttttttaaga aacacggtaa gggaaaagct
     3301 agtagttgca aggcttgcgt ggactattta ctaaataagc ctgacgacac cgcccaaatc
     3361 ctgcagggcg atccccgact atcacaaagt attgctgata gtcttgattt taataacact
     3421 tacaccgcag gttgcttgtc ttttgaagaa agcgacctac ccgaaataca aaagcgtgag
     3481 attatggcac gctttgaaaa ggcaatgttt gcagggcttg agcctgaaca atataacatt
     3541 gcgtgggtac aacacaccga caaaggaagg cttgagctga atttcgttat cccaaacgta
     3601 gagatgacaa gcggaaaacg cctacaacct tactacgaca gggcagaccg cccacttgct
     3661 gaaaacttca agcaggtaat caaccacgaa tacagcctaa gcgacccaaa caaccctata
     3721 aagcaaaaaa acctgattga ccgcaaagac ctaccaaccg acaaaaaaca ggctttacag
     3781 gcgattacgg acggtttaac agctttagcg aacgcagggc agataaacga ccgacaggac
     3841 gttataaatg ccctagaacg tgcaggtttt gaaattgcac gcattacgcc aaaaaaccta
     3901 tcaatcaaga ctgacggaca gaatttaaga ttaaaagggg ctttctatga gcaagatttt
     3961 agatttagca cagacctttc agcagacatc acagaaagag ctagagagta caagcgagat
     4021 agtgcagaac gctatcaaac ggcacgagca aaacttgata cagcagttac agcacgcagg
     4081 gaacaattta gccgaaaata tccaaatcga gcaggcgaaa ttgataaaaa ataccgtgag
     4141 aatgtatcgc ttgccgaccc taatcgcctt gacgatatta accttgatag ccataaccac
     4201 tccatatccg gatctttaat aagttgttta aaaattttgg tgaatttatc caattttgtc
     4261 catcttctaa aagttttaaa aacagtatta tactttccaa agcattctgg taaatatcgc
     4321 catgggcagc ctgttttcag gcgatataag attcctgtga aaatatttct taaatttggt
     4381 ttgtcataga tattcaaatc taggataata ggctttagtt tcaaccaaaa tcgaattctt
     4441 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4501 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4561 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4621 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4681 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4741 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4801 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4861 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     4921 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4981 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5041 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5101 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5161 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     5221 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5281 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5341 ctggagccgg tgagcgtgga tctcgcggta tcattgcagc actggggcca gatggtaagc
     5401 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5461 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5521 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5581 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5641 cgtcagaccc caattacacg ccactggctg tgcttgctgg ggtgacggtg gcaacggtgg
     5701 cggccttgct gggctatcgc gttggaaaga aacgagggaa aggggactga taaaccggtc
     5761 ttagcccctc cccttggtgt ccaaccgctc tgtaggatcg aattcctgca gcccgtcatt
     5821 aagtaatgtt tgaagcataa gaatagagag ttagtattat tttttgcgat tacattttac
     5881 tactctctat ttttttattt caaatgttca acagacccta ttatagaagc ccccatgaat
     5941 aaatcgctca tcattttcgg catcgtcaac ataacctcgg acagtttctc cgatggaggc
     6001 cggtatctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     6061 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     6121 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     6181 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     6241 caagctttct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6301 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6361 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6421 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6481 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6541 tttatagtcc tgtcgggttt cgc
//
