LOCUS       sequence-130923-        6439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces E. coli acetyl-CoA:oxalate CoA-transferase (yfdE) with 
            N-terminal His6 tag (H6YfdE).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6439)
  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 6439)
  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: pJK593
FEATURES             Location/Qualifiers
     source          1..6439
                     /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"
     CDS             complement(1313..1330)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1340..1357)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1376..1398
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1413..1437
                     /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     complement(1437..1456)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1438..1456)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1540..1559)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1765..1842
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1843..2925
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1862..1881)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3734..3925
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3919..3941)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4027..4169
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4082..4101
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4184..4201)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4355..4943)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4435..4454)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5054..5073)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5065..5880
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             5065..5880
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5138..5157)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5973..6428)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6110..6131)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6322..6341
                     /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 atggcgattt 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 tggctgccgc
     1321 gcggcaccag gccgctgctg tgatgatgat gatgatggct gctgcccatg gtatatctcc
     1381 ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca caattcccct
     1441 atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc ggacgcatcg
     1501 tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg
     1561 atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg
     1621 tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg
     1681 cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc
     1741 ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt tcgcggtatg
     1801 gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta
     1861 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
     1921 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
     1981 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
     2041 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
     2101 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
     2161 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
     2221 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
     2281 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt
     2341 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
     2401 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
     2461 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
     2521 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
     2581 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
     2641 gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta
     2701 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
     2761 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
     2821 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     2881 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     2941 tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga gagccttcaa
     3001 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
     3061 cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca ttttcggcga
     3121 ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt
     3181 gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca
     3241 ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc tgtcgttgag
     3301 gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga
     3361 gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt
     3421 cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat
     3481 gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt
     3541 aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc
     3601 cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat
     3661 cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta
     3721 cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag
     3781 aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga
     3841 catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt
     3901 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     3961 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     4021 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
     4081 gcggcatcag agcagattgt actgagagtg caccatatat gcggtgtgaa ataccgcaca
     4141 gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc
     4201 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4261 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4321 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4381 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4441 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4501 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4561 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4621 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4681 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4741 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     4801 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4861 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4921 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4981 agtggaacga aaactcacgt taagggattt tggtcatgaa caataaaact gtctgcttac
     5041 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc ttgctctagg
     5101 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     5161 gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg
     5221 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     5281 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     5341 gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt attagaagaa
     5401 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     5461 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     5521 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     5581 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     5641 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     5701 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     5761 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     5821 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     5881 gaattaattc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5941 tccgcgcaca tttccccgaa aagtgccacc tgaaattgta aacgttaata ttttgttaaa
     6001 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     6061 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     6121 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     6181 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     6241 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     6301 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     6361 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     6421 gggcgcgtcc cattcgcca
//
