LOCUS       sequence-130928-        4798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Acetobacter aceti 1023 acetyl-CoA:oxalate CoA-transferase 
            (UctC) with Thr2>Ala mutant (UctC-T2A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4798)
  AUTHORS   Mullins EA, Francois JA, Kappock TJ
  TITLE     A specialized citric acid cycle requiring succinyl-coenzyme A 
            (CoA):acetate CoA-transferase (AarC) confers acetic acid resistance 
            on the acidophile Acetobacter aceti.
  JOURNAL   J Bacteriol. 2008 Jul;190(14):4933-40. doi: 10.1128/JB.00405-08. 
            Epub 2008 May 23.
  PUBMED    18502856
REFERENCE   2  (bases 1 to 4798)
  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"; doi: 
            "10.1128/JB.00405-08"; journalName: "J Bacteriol"; date: "2008-07"; 
            volume: "190"; issue: "14"; pages: "4933-40"
COMMENT     Sequence Label: pJK360
FEATURES             Location/Qualifiers
     source          1..4798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1379..1401
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1432..1451)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1433..1451)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1960..2151
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2145..2167)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2253..2395
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2308..2327
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2410..2427)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2581..3169)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2661..2680)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3340..4131,4132..4200))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3963..3982
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4201..4305)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4332..4787)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4469..4490)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4681..4700
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcagg cgtcattttg cctctggtgc ggcaaattcc ttacgcaacg
      241 ccgcgccctg ctcatccagc ttaggggcgc cgggccgttc atgtgggtca tcatacccac
      301 tgattttcac cgggttaccg ggcatcatca ccccgccaga tttaatcagc atattccgcg
      361 cggctgtttg tggcagattt gccgcctctg ccacgtccag caacggccca acaggcacac
      421 cggctttgtg aataacatcc agccaatgcg ccgctggctg tttggaaagt gctttttcca
      481 atgcagcttt cagcgcatca ttattggcca ttctatcatg attttcgaca aaacgcgggt
      541 ctgtcgccag ttcaggcgca ccaatcacat cgcaaagcgc cttaaaaaga tggtcattcc
      601 cacaacagat gacaaaaggc ttgtcctgcg cctgaaacac atcaaaaggc gccatataag
      661 gatggcgatt gccaatacgc gcaggggctt tgcctgtagc agaccagcac atcagagcat
      721 gctgcaaaaa tgccaaggta ccatcaaaca ttgaaacatc aatgtgtgcc cctttccctg
      781 tgcgctcacg cgcataaagc gcactggcaa tgccgcagaa catgaaaacc ccaccgcaca
      841 gatcagaaag cgatgtgccc cctacccgcg ttggcggacc atcgggaaag cccgtggcca
      901 tcatgatgcc gctcatggcc tgcacaatag tatcgtaggc aggataatgt gcgagcgggc
      961 ctgtctgacc gaaccctgat gaagatgcat aaatcaaccg tggattgatc tttgccagtt
     1021 cctcataatt gaaacccaaa cggtccatca cgccggggcg gaagttttcc gccagcacat
     1081 cggccttacg caccatttcc agaaaaatcg ctcgatcgcc ctcatccttg aggttaagga
     1141 caatgctttc ctttcccctg tttacaaaag agaagtaaag cgattggtct ccaacatatg
     1201 ggccataggt acgcgtatca tccccatgcc ccggtggctc gatcttgatt gtacgagccc
     1261 ccagatctgt tagaattgtt gtgccgaacg gaccattcag cacatgtgtc agatcaataa
     1321 cgaggagccc atcaaagggc cccttcggtt ttgctggcgt ggtttcagcc atggtatatc
     1381 tccttcttaa agttaaacaa aattatttct agagggaaac cgttgtggtc tccctatagt
     1441 gagtcgtatt aatttcgcgg gatcgagatc tcgggcagcg ttgggtcctg gccacgggtg
     1501 cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt
     1561 tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct
     1621 gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg
     1681 cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta
     1741 ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt
     1801 ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg
     1861 gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt
     1921 acccccatga acagaaatcc cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc
     1981 gcccttaaca tggcccgctt tatcagaagc cagacattaa cgcttctgga gaaactcaac
     2041 gagctggacg cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag
     2101 ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     2161 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     2221 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     2281 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     2341 atatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc
     2401 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2461 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2521 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2581 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2641 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2701 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2761 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2821 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2881 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2941 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3001 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     3061 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3121 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3181 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3241 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3301 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3361 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3421 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3481 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3541 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3601 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg
     3661 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3721 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3781 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3841 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3901 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3961 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4021 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4081 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4141 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4201 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4261 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4321 agtgccacct gaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     4381 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     4441 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     4501 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     4561 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     4621 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     4681 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     4741 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgcca
//
