LOCUS       sequence-256047-        7038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A recombinant plasmid expressing fum (fumarase) gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7038)
  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 7038)
  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: "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: pFUM
FEATURES             Location/Qualifiers
     source          1..7038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     214..231
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2161..2178)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2161..2177)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2170..2192)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4928..4946)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5014..5118
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5119..5187,5188..5979)
                     /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(5337..5356)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    6590..6611
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6626..6631,6632..6649,6650..6656)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6664..6680
                     /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     6669..6691
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6688..6704
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6688..6704
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6999..7018
                     /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 tgtttgagtg aaaacagccg tgttttttca tttaccgcca taaaaatttg aaacggatca
      361 caaatcatga aaaaaatacg ttcaaattag aactaattat cgaaaatttg atctagttaa
      421 cattttttag gtataaatag ttttaaaata gatctagttt ggatttttaa ttttaaatta
      481 tcaatgaggt gaagtatgac atttcgtatt gaaaaagaca ctatgggcga ggttcaagtt
      541 cctgcggata aatattgggc cgcacaaacg gaacgttcac gcaataactt taaaatcgga
      601 cctgcagcgt caatgccaca cgaaattatt gaagcattcg gttatttgaa aaaagcggcc
      661 gcttatgcca acgcggattt aggcgtattg ccggctgaaa aacgcgattt gatcgcacaa
      721 gcctgcgatg aaattctggc tcgtaaatta gatgatcaat tcccgttagt tatctggcaa
      781 accggttcgg gtacacaatc caatatgaat ctgaacgaag ttatcgctaa ccgcgcacat
      841 gtgattaacg gtggtaaatt aggtgaaaaa tctattattc atccaaacga cgatgtaaac
      901 aaatctcaat cttcaaacga tacttatccg acagcaatgc acattgccac atataagaaa
      961 gtggttgaag caacgattcc ggccatcgaa cgtttacaaa aaaccttagc ggcgaaatcc
     1021 gaagaattca aagatgtggt gaaaatcggc cgtacgcact taatggatgc caccccgttg
     1081 acattgggtc aggaattcag cggttatgct gcacaattaa gtttcggttt agcggcaatc
     1141 aaaaatacct taccgcattt acgccaactg gcattaggcg gtacggcagt gggtaccggt
     1201 ttaaatacac ctaaaggcta tgatgtaaaa gtagcggaat atatcgccaa attcaccggc
     1261 ttgccgttta ttaccgccga aaacaaattt gaagcattag caacacatga cgctatcgtt
     1321 gaaactcacg gcgcattaaa acaagttgcg atgtccttat tcaaaattgc aaatgatatc
     1381 cgtttattgg cttcaggtcc tcgttctggt atcggtgaaa ttttaattcc tgaaaacgaa
     1441 ccgggttcat ccatcatgcc gggtaaagtt aatccgaccc aatgcgaagc gatgacaatg
     1501 gttgccgcac aagtattagg taacgatacc actatttcat ttgccggttc gcaaggtcat
     1561 ttcgaattga acgtattcaa accggttatg gcggcaaatt tcctgcaatc cgctcaatta
     1621 atcgcagatg tttgcatttc tttcgacgag cactgtgcaa gcggcattaa accaaatacg
     1681 ccgcgcattc aacacttact tgaaagttca ttaatgttag tgaccgcatt aaatactcat
     1741 atcggttatg aaaatgcggc gaaaattgcg aaaactgcgc acaaaaacgg tacaacatta
     1801 cgtgaagagg ctatcaactt aggtttagtg tccgccgaag atttcgataa atgggttcgt
     1861 ccggaagata tggttggcag cttaaaataa gcggcagata tgaataaaaa ttgaaaggcg
     1921 acagaagtcg cctttttttt ctggctagag ttggtataat tttgccaatt cgtttatctt
     1981 ggattcttta tgaaattgaa atcacttttt attgccattt tatgcttaat tcccaccgca
     2041 ctttcggcga attggctgca agtgggaaaa gcagattata actggggtcc gttccacgtt
     2101 tatactatat cagtattcac cgaagacggt cagggatccc cgggtaccga gctcgaattc
     2161 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     2221 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     2281 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc cagatgaagc agagcgcgcc
     2341 ttcgctgtcc tattcaatgt attggggatc gaagcccccg accgcgaacg ccttgccttc
     2401 tatctgcgat tggaccctct gacttggggt tgatgttcat gccgcctgtt tttcctgctc
     2461 attggcacgt ttcgcaacct gttctcattg cggacacctt ttccagcctc gtttggaaag
     2521 tttcattgcc agtaatacca atgctttaga aagaaaaagg gatcgaactt ttgacattcg
     2581 atcccttttt ctgtaatctg tttcgtgcgt tctttgctaa gatacagacc ctagacaagt
     2641 catatcttag caaagggtag ctagtaatgc aagagattgc gaagcgtccc tactaccaaa
     2701 aaaccattca acgacgtaaa cagacaaacg caaaccttaa attagacggt cttcagctcg
     2761 gacttcggaa gaataaacag gcgtagaagt gataacgttc ttaatacgaa aattaagctc
     2821 tgtctccgtt tcgtgctacg gttagaaagg cgaaagcccc aagaaataca agcacacctg
     2881 ataagcgaga tttaaggata acagcgaaat tcaatagggt ctgaatttcc aaactaggtt
     2941 aaatgccacg acgttttatt gttgccccat tcaagcaaca tttgagaacc gaatagaaat
     3001 cttttagtaa aaagcgttct tttttgggtc agcggttaat gtggacggtt taacggtttt
     3061 tcccctgcgg gtcgtattgg aaagccattg aaaagctgat ggataactct gcgagttacc
     3121 cacgagcttt ccaacagctt tccaacacta aaaacctacc gcccacaata accacttccc
     3181 taataataaa atttttttat ttttattttg gttcaaaggc tcacgatgtt cgcctaataa
     3241 aacgaagtcg cctatcggct ccgctgattt ttatatatca ctctcggggc ttttggtgta
     3301 ctattgtctt ttgtaatagc aaggacacaa aaagggtact cttcgagttt cctttttgac
     3361 cttgcaaaag ggctttgccc ccttgacccc cgaccgcttt cagcggtcaa aatagaagaa
     3421 cggacaccat tatgaaacgt gagaaagaga taaaaatcag gctcaccgaa aacgagtatc
     3481 aagccttgtt agagagaaaa acgaaagcaa ggcttgcgga gtgggttcgg gaagttgccc
     3541 tggaacagca acctaagcga cagccgaaag taatcgaccc tgcgttactg ttcgagctga
     3601 accgcatagg cgtaaacctg aaccaaatcg cccgacaatg caacagtcaa aagccgagca
     3661 ttgaccttgt tagcgtgttg gcgaccttgc gagaaattga aaaaaatctc aaaaaattgc
     3721 gagaattgag cctatgatcg ttaaattttt taagaaacac ggtaagggaa aagctagtag
     3781 ttgcaaggct tgcgtggact atttactaaa taagcctgac gacaccgccc aaatcctgca
     3841 gggcgatccc cgactatcac aaagtattgc tgatagtctt gattttaata acacttacac
     3901 cgcaggttgc ttgtcttttg aagaaagcga cctacccgaa atacaaaagc gtgagattat
     3961 ggcacgcttt gaaaaggcaa tgtttgcagg gcttgagcct gaacaatata acattgcgtg
     4021 ggtacaacac accgacaaag gaaggcttga gctgaatttc gttatcccaa acgtagagat
     4081 gacaagcgga aaacgcctac aaccttacta cgacagggca gaccgcccac ttgctgaaaa
     4141 cttcaagcag gtaatcaacc acgaatacag cctaagcgac ccaaacaacc ctataaagca
     4201 aaaaaacctg attgaccgca aagacctacc aaccgacaaa aaacaggctt tacaggcgat
     4261 tacggacggt ttaacagctt tagcgaacgc agggcagata aacgaccgac aggacgttat
     4321 aaatgcccta gaacgtgcag gttttgaaat tgcacgcatt acgccaaaaa acctatcaat
     4381 caagactgac ggacagaatt taagattaaa aggggctttc tatgagcaag attttagatt
     4441 tagcacagac ctttcagcag acatcacaga aagagctaga gagtacaagc gagatagtgc
     4501 agaacgctat caaacggcac gagcaaaact tgatacagca gttacagcac gcagggaaca
     4561 atttagccga aaatatccaa atcgagcagg cgaaattgat aaaaaatacc gtgagaatgt
     4621 atcgcttgcc gaccctaatc gccttgacga tattaacctt gatagccata accactccat
     4681 atccggatct ttaataagtt gtttaaaaat tttggtgaat ttatccaatt ttgtccatct
     4741 tctaaaagtt ttaaaaacag tattatactt tccaaagcat tctggtaaat atcgccatgg
     4801 gcagcctgtt ttcaggcgat ataagattcc tgtgaaaata tttcttaaat ttggtttgtc
     4861 atagatattc aaatctagga taataggctt tagtttcaac caaaatcgaa ttcttgaaga
     4921 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4981 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5041 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5101 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5161 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5221 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5281 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5341 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     5401 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5461 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5521 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5581 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5641 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     5701 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5761 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5821 gccggtgagc gtggatctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5881 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5941 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6001 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6061 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6121 gaccccaatt acacgccact ggctgtgctt gctggggtga cggtggcaac ggtggcggcc
     6181 ttgctgggct atcgcgttgg aaagaaacga gggaaagggg actgataaac cggtcttagc
     6241 ccctcccctt ggtgtccaac cgctctgtag gatcgaattc ctgcagcccg tcattaagta
     6301 atgtttgaag cataagaata gagagttagt attatttttt gcgattacat tttactactc
     6361 tctatttttt tatttcaaat gttcaacaga ccctattata gaagccccca tgaataaatc
     6421 gctcatcatt ttcggcatcg tcaacataac ctcggacagt ttctccgatg gaggccggta
     6481 tctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     6541 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     6601 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     6661 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     6721 tttctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6781 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6841 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6901 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6961 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7021 agtcctgtcg ggtttcgc
//
