LOCUS       sequence-256043-        7226 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a recombinant plasmid, expressing pckA PEP-carboxykinase) gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7226)
  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 7226)
  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: pPCK
FEATURES             Location/Qualifiers
     source          1..7226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     214..231
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2349..2366)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2349..2365)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2358..2380)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5116..5134)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5202..5306
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5307..5375,5376..6167)
                     /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(5525..5544)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    6778..6799
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6814..6819,6820..6837,6838..6844)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6852..6868
                     /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     6857..6879
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6876..6892
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6876..6892
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7187..7206
                     /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 tgacttaaac aaactcgtta aagaacttaa tgacttaggg
      541 cttaccgatg ttaaggaaat tgtgtataac ccgagttatg aacaactttt cgaggaagaa
      601 accaaaccgg gtttggaggg tttcgataaa gggacgttaa ccacgcttgg cgcggttgcc
      661 gtcgatacgg ggatttttac cggtcgttca ccgaaagata aatatatcgt ttgcgatgaa
      721 actacgaaag acaccgtttg gtggaacagc gaagcggcga aaaacgataa caaaccgatg
      781 acgcaagaaa cttggaaaag tttgagagaa ttagtggcga aacaactttc cggtaaacgt
      841 ttattcgtgg tagaaggtta ctgcggcgcc agtgaaaaac accgtatcgg tgtgcgtatg
      901 gttactgaag tggcatggca ggcgcatttt gtgaaaaaca tgtttatccg accgaccgat
      961 gaagagttga aaaatttcaa agcggatttt accgtgttaa acggtgctaa atgtactaat
     1021 ccgaactgga aagaacaagg tttgaacagt gaaaactttg tcgctttcaa tattaccgaa
     1081 ggtattcagc ttatcggcgg tacttggtac ggcggtgaaa tgaaaaaagg tatgttctca
     1141 atgatgaact acttcctgcc gttaaaaggt gtggcttcca tgcactgttc cgccaacgta
     1201 ggtaaagacg gtgacgtggc tattttcttc ggtttatccg gtacgggtaa aacaacgctt
     1261 tcgaccgatc ctaaacgcca attaatcggt gatgacgaac acggttggga tgaatccggc
     1321 gtatttaact ttgaaggcgg ttgttacgcg aaaaccatta acttatctca agaaaacgaa
     1381 ccggatattt acggcgcaat ccgtcgtgac gcattattag aaaacgtcgt ggttcgtgca
     1441 gacggttccg ttgactttga cgacggttca aaaacagaaa atacccgtgt ttcatatccg
     1501 atttaccaca tcgacaacat cgttcgtccg gtatcgaaag ccggtcatgc aaccaaagtg
     1561 attttcttaa ccgcggacgc attcggcgta ttgccgccgg tttcaaaact gactccggaa
     1621 caaaccgaat actacttctt atccggcttt actgcaaaat tagcgggtac ggaacgcggc
     1681 gtaaccgaac cgactccgac attctcggcc tgtttcggtg cggcattctt aagcctgcat
     1741 ccgattcaat atgcggacgt gttggtcgaa cgcatgaaag cctccggtgc ggaagcttat
     1801 ttggtgaaca ccggttggaa cggcacgggt aaacgtattt caatcaaaga tacccgcggt
     1861 attatcgatg cgattttgga cggttcaatc gaaaaagcgg aaatgggcga attgccaatc
     1921 tttaatttag cgattcctaa agcattaccg ggtgttgatc ctgctatttt ggatccgcgc
     1981 gatacttacg cagacaaagc gcaatggcaa gttaaagcgg aagatttggc aaaccgtttc
     2041 gtgaaaaact ttgtgaaata tacggcgaat ccggaagcgg ctaaattagt tggcgccggt
     2101 ccaaaagcat aaaactgtaa aagcatagta tgtgcatgat tcggtaaact accgaataaa
     2161 atctgaaaaa taaaggctga gtatttccac tcagcctttt gttttggaaa ttagaagttg
     2221 tttagaaaga taaacggcgc ggcttattcg tccgtattat ttggggacat aaaaaacccg
     2281 ttgataaaca acgggtttgt catcttaaca ggttattagg ccgccggcac ggtaccgagc
     2341 tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     2401 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     2461 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcca gatgaagcag
     2521 agcgcgcctt cgctgtccta ttcaatgtat tggggatcga agcccccgac cgcgaacgcc
     2581 ttgccttcta tctgcgattg gaccctctga cttggggttg atgttcatgc cgcctgtttt
     2641 tcctgctcat tggcacgttt cgcaacctgt tctcattgcg gacacctttt ccagcctcgt
     2701 ttggaaagtt tcattgccag taataccaat gctttagaaa gaaaaaggga tcgaactttt
     2761 gacattcgat ccctttttct gtaatctgtt tcgtgcgttc tttgctaaga tacagaccct
     2821 agacaagtca tatcttagca aagggtagct agtaatgcaa gagattgcga agcgtcccta
     2881 ctaccaaaaa accattcaac gacgtaaaca gacaaacgca aaccttaaat tagacggtct
     2941 tcagctcgga cttcggaaga ataaacaggc gtagaagtga taacgttctt aatacgaaaa
     3001 ttaagctctg tctccgtttc gtgctacggt tagaaaggcg aaagccccaa gaaatacaag
     3061 cacacctgat aagcgagatt taaggataac agcgaaattc aatagggtct gaatttccaa
     3121 actaggttaa atgccacgac gttttattgt tgccccattc aagcaacatt tgagaaccga
     3181 atagaaatct tttagtaaaa agcgttcttt tttgggtcag cggttaatgt ggacggttta
     3241 acggtttttc ccctgcgggt cgtattggaa agccattgaa aagctgatgg ataactctgc
     3301 gagttaccca cgagctttcc aacagctttc caacactaaa aacctaccgc ccacaataac
     3361 cacttcccta ataataaaat ttttttattt ttattttggt tcaaaggctc acgatgttcg
     3421 cctaataaaa cgaagtcgcc tatcggctcc gctgattttt atatatcact ctcggggctt
     3481 ttggtgtact attgtctttt gtaatagcaa ggacacaaaa agggtactct tcgagtttcc
     3541 tttttgacct tgcaaaaggg ctttgccccc ttgacccccg accgctttca gcggtcaaaa
     3601 tagaagaacg gacaccatta tgaaacgtga gaaagagata aaaatcaggc tcaccgaaaa
     3661 cgagtatcaa gccttgttag agagaaaaac gaaagcaagg cttgcggagt gggttcggga
     3721 agttgccctg gaacagcaac ctaagcgaca gccgaaagta atcgaccctg cgttactgtt
     3781 cgagctgaac cgcataggcg taaacctgaa ccaaatcgcc cgacaatgca acagtcaaaa
     3841 gccgagcatt gaccttgtta gcgtgttggc gaccttgcga gaaattgaaa aaaatctcaa
     3901 aaaattgcga gaattgagcc tatgatcgtt aaatttttta agaaacacgg taagggaaaa
     3961 gctagtagtt gcaaggcttg cgtggactat ttactaaata agcctgacga caccgcccaa
     4021 atcctgcagg gcgatccccg actatcacaa agtattgctg atagtcttga ttttaataac
     4081 acttacaccg caggttgctt gtcttttgaa gaaagcgacc tacccgaaat acaaaagcgt
     4141 gagattatgg cacgctttga aaaggcaatg tttgcagggc ttgagcctga acaatataac
     4201 attgcgtggg tacaacacac cgacaaagga aggcttgagc tgaatttcgt tatcccaaac
     4261 gtagagatga caagcggaaa acgcctacaa ccttactacg acagggcaga ccgcccactt
     4321 gctgaaaact tcaagcaggt aatcaaccac gaatacagcc taagcgaccc aaacaaccct
     4381 ataaagcaaa aaaacctgat tgaccgcaaa gacctaccaa ccgacaaaaa acaggcttta
     4441 caggcgatta cggacggttt aacagcttta gcgaacgcag ggcagataaa cgaccgacag
     4501 gacgttataa atgccctaga acgtgcaggt tttgaaattg cacgcattac gccaaaaaac
     4561 ctatcaatca agactgacgg acagaattta agattaaaag gggctttcta tgagcaagat
     4621 tttagattta gcacagacct ttcagcagac atcacagaaa gagctagaga gtacaagcga
     4681 gatagtgcag aacgctatca aacggcacga gcaaaacttg atacagcagt tacagcacgc
     4741 agggaacaat ttagccgaaa atatccaaat cgagcaggcg aaattgataa aaaataccgt
     4801 gagaatgtat cgcttgccga ccctaatcgc cttgacgata ttaaccttga tagccataac
     4861 cactccatat ccggatcttt aataagttgt ttaaaaattt tggtgaattt atccaatttt
     4921 gtccatcttc taaaagtttt aaaaacagta ttatactttc caaagcattc tggtaaatat
     4981 cgccatgggc agcctgtttt caggcgatat aagattcctg tgaaaatatt tcttaaattt
     5041 ggtttgtcat agatattcaa atctaggata ataggcttta gtttcaacca aaatcgaatt
     5101 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     5161 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5221 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5281 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5341 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5401 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5461 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     5521 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     5581 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     5641 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5701 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     5761 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5821 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     5881 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5941 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6001 aatctggagc cggtgagcgt ggatctcgcg gtatcattgc agcactgggg ccagatggta
     6061 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6121 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6181 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6241 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6301 gagcgtcaga ccccaattac acgccactgg ctgtgcttgc tggggtgacg gtggcaacgg
     6361 tggcggcctt gctgggctat cgcgttggaa agaaacgagg gaaaggggac tgataaaccg
     6421 gtcttagccc ctccccttgg tgtccaaccg ctctgtagga tcgaattcct gcagcccgtc
     6481 attaagtaat gtttgaagca taagaataga gagttagtat tattttttgc gattacattt
     6541 tactactctc tattttttta tttcaaatgt tcaacagacc ctattataga agcccccatg
     6601 aataaatcgc tcatcatttt cggcatcgtc aacataacct cggacagttt ctccgatgga
     6661 ggccggtatc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     6721 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     6781 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     6841 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     6901 cgccaagctt tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6961 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7021 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7081 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7141 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7201 atctttatag tcctgtcggg tttcgc
//
