LOCUS       sequence-130931-        5108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Acetobacter aceti 1023 succinyl-CoA:acetate CoA-transferase
            with C-terminal His6 tag (AarCH6).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5108)
  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 5108)
  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: pJK385
FEATURES             Location/Qualifiers
     source          1..5108
                     /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             1689..1711
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1742..1761)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1743..1761)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2270..2461
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2455..2477)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2563..2705
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2618..2637
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2720..2737)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2891..3479)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2971..2990)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3650..4441,4442..4510))
                     /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     4273..4292
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4511..4615)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4642..5097)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4779..4800)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4991..5010
                     /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 gagtgatgat gggagcatgg
      181 tgcccgtatc aataaaccgc tgatgccaag acagagcttc cgtcagcaga tgcggtgtgt
      241 gcttaccaaa ggaattttta agcgcacgat caaaataatc ctgcaacatc ggccggtaat
      301 ccggatgcgc acatttggaa ataatttcac gcgcacgttg caccggtgaa agcccacgca
      361 gatccgccag cccctgctcg gttacaaaaa tctgcgcatc ctgcatgatg tggtccacat
      421 gcgcagccat cggcacaatg gctgaaatct tgccaccttt ggctgtggag ggagacagga
      481 agatggacag gtaggaactc cgggcaaaat cacccgatcc cccaatacca ttcatcattt
      541 tggaacccat cacgcgggta gaattcacgt ttccgtaaat atccgcctca atcatgccgt
      601 tcatggcaat gcagcctaaa cggcggataa tgccggggct gttgctgaca tcctgctggc
      661 gcagaatgat cttgctgcgg aagaaatcca tccggttatt gatttcttcc gcagcttccg
      721 ggctcagcga gaaagacgag gcagaggcaa tacgcatacg gccagaatcc agcatggcca
      781 acatgccatc ctgaatcacc tcactatagc ccaccagatt ttcaaatggg ccttccttca
      841 gcccttccag cacggcgttg gccacattgc ccacaccaga ctggagcggc agaagtgagg
      901 gcggcaaacg gttctgtttg acttcatgcc caaagaaatc aagcaaatag cccgcaatag
      961 cttttgctgt ttcatccggc gcggcaaacg gggcatttct gtcccggtca ttcgtgcgta
     1021 caatggcagc aatcttgtcc gggttcaccc gtaaaaccgg gccacccacg cgctgatctg
     1081 cccgtacaat cggcacaata tcacgcgtag gcacgccact tacgttgcca tcccagatat
     1141 catggatgcc ttccagcatc ggattctgcc attcatttac ctcgataatg accttttcgg
     1201 ccagattgag gaatgtctgg gaattgccga cagaagatgt gggcacaatg ccgccatctt
     1261 ccgtaatggc tgtggcttcc accagcgcaa tattgaactt gccgtaattg ccctgcacgg
     1321 cacggccagc cacctgcccc agatggttat caaaatattc cgtttcaccc gcattaatac
     1381 ggttgcgcat ggtggcatcc gtattgaatg gcgagcggaa atacaccccg ttggccttgg
     1441 ccagctcacc atccagctga gggcccgtgg atgcgcccgt aatcaggctg atctggtatt
     1501 tttcgccccg atcatgcgca gcttccatcc gctgggccag cgccttgggt acttccttgg
     1561 ggtaacctgc cccggtaaaa ccgctggtcc ccaccacgtc tccgtgcttg atcaactcgg
     1621 atgcggtttc tgccggacat actttactgc gcagcgcgac gttccgaatg cgctctgtca
     1681 tatgtatatc tccttcttaa agttaaacaa aattatttct agagggaaac cgttgtggtc
     1741 tccctatagt gagtcgtatt aatttcgcgg gatcgagatc tcgggcagcg ttgggtcctg
     1801 gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg
     1861 ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg
     1921 caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag
     1981 tctggaaacg cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg
     2041 ctgctggcta ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg
     2101 agtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc
     2161 cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat
     2221 cggtatcatt acccccatga acagaaatcc cccttacacg gaggcatcag tgaccaaaca
     2281 ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc cagacattaa cgcttctgga
     2341 gaaactcaac gagctggacg cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca
     2401 cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
     2461 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     2521 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
     2581 acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg
     2641 agagtgcacc atatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2701 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2761 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2821 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2881 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2941 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3001 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3061 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3121 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3181 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3241 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3301 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     3361 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3421 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3481 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3541 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3601 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3661 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3721 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3781 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3841 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3901 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3961 ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4021 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4081 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4141 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4201 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4261 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4321 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4381 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4441 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4501 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4561 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4621 ttccccgaaa agtgccacct gaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa
     4681 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4741 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4801 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4861 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4921 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4981 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     5041 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     5101 attcgcca
//
