LOCUS       sequence-130921-        4736 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces E. coli acetyl-CoA:oxalate CoA-transferase (yfdE) with 
            Arg232>His mutant (YfdE-R232H).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4736)
  AUTHORS   Mullins EA, Sullivan KL, Kappock TJ
  TITLE     Function and X-ray crystal structure of Escherichia coli YfdE.
  JOURNAL   PLoS One. 2013 Jul 23;8(7):e67901. doi: 
            10.1371/journal.pone.0067901. Print 2013.
  PUBMED    23935849
REFERENCE   2  (bases 1 to 4736)
  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: "PLoS One.";
            date: "2013-07-23"; volume: "8(7):e67901. doi"; pages: " 
            10.1371/journal.pone.0067901. Print 2013"
COMMENT     Sequence Label: pJK560
FEATURES             Location/Qualifiers
     source          1..4736
                     /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             1317..1339
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1370..1389)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1371..1389)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1898..2089
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2083..2105)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2191..2333
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2246..2265
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2348..2365)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2519..3107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2599..2618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3278..4069,4070..4138))
                     /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     3901..3920
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4139..4243)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4270..4725)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4407..4428)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4619..4638
                     /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 gagttatgat gagaactcct
      181 ggcgaatttg ttccccatgc tggtcgagcg ttgccgctcc cggcataaca tgcgggtccg
      241 cgcagccgct gatttttatc ggattacccg gcatcattat tcccccggct tcaatcaaca
      301 tatttctcgc ctgagtttgt ggcaatttaa tggcctcagc cacacttaat aacggcgcga
      361 cgggtacacc aacttcatgt attctggcta accaaacttc agctgcctgc gtttttaacg
      421 tccgctcaat atattgttta agaatagcct ggttttgtac gcgtaaaata ttgctgctaa
      481 atcggggatc attaaccagt tccgtaagct ccagtgcctg gcataacgca gaaaaaagct
      541 tgtcattacc acaacaaatc gtaatcggct tatcctgagt attgaaaaca tcaaaaggtg
      601 ccatgtaggg atggtgattt cccagacgtt gtggtgactt cccagttgcg atatatgcca
      661 tcagaccatg ctccagaaaa ctcagcgtgg catcaaacat cgctatatcg acatgcgccc
      721 ctctctggct cttttcgcga ccataaagtg cactcactat tccgctgaat aaatagacac
      781 cgccgcatag atccgcaaga gatgtaccaa cgcgcactgg cggagcatca ggatatcctg
      841 tttccatcat tatcccgctc attgcctgaa tgatggtatc gtaggcagga gcatctttta
      901 gcggaccggt atgtccgaaa cctgacgatg aagcatatat gaggcgcggg ttgatttctt
      961 gaagcgtttc ccatgaaaac cccagttttt ccattgtacc tgggcgaaaa ttctcagcta
     1021 atacatcagc ttgtttgagc atatttataa atatactttt atcgtgatca ttctttaaat
     1081 caagaaccac actctctttg ccatgattaa taaaactgta atagagtgac tgtccatcca
     1141 catagggacc aaatgtgcgg gtatcatcac catgacccgg cggctcaact ttaattaccc
     1201 ttgcgcccat attacaaaga agttgagttc cgaaaggtcc attaaggaca tgtgtcatat
     1261 cgataactaa taagccttca aacggccctt tgctttcatt atttgtcata tgtatatctc
     1321 cttcttaaag ttaaacaaaa ttatttctag agggaaaccg ttgtggtctc cctatagtga
     1381 gtcgtattaa tttcgcggga tcgagatctc gggcagcgtt gggtcctggc cacgggtgcg
     1441 catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta
     1501 gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc
     1561 gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg
     1621 gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc
     1681 ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct
     1741 ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc
     1801 atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac
     1861 ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc
     1921 ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga
     1981 gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct
     2041 ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     2101 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     2161 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     2221 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     2281 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     2341 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2401 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2461 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2521 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2581 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2641 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2701 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2761 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2821 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2881 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2941 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3001 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3061 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3121 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3181 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3241 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3301 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3361 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3421 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3481 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3541 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca
     3601 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3661 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3721 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3781 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3841 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3901 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3961 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4021 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4081 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4141 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4201 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4261 tgccacctga aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     4321 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     4381 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     4441 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     4501 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     4561 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     4621 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     4681 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgcca
//
