LOCUS       sequence-127060-        5943 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Produces E coli formyl-CoA:oxalate CoA-transferase (H6YfdW) in E. 
            coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5943)
  AUTHORS   Mullins EA, Starks CM, Francois JA, Sael L, Kihara D, Kappock TJ
  TITLE     Formyl-coenzyme A (CoA):oxalate CoA-transferase from the acidophile 
            Acetobacter aceti has a distinctive electrostatic surface and 
            inherent acid stability.
  JOURNAL   Protein Sci. 2012 May;21(5):686-96. doi: 10.1002/pro.2054. Epub 2012
            Mar 29.
  PUBMED    22374910
REFERENCE   2  (bases 1 to 5943)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1002/pro.2054"; 
            journalName: "Protein Sci"; date: "2012-05"; volume: "21"; issue: 
            "5"; pages: "686-96"
COMMENT     Sequence Label: pJK597
FEATURES             Location/Qualifiers
     source          1..5943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1285..1302)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1312..1329)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1348..1370
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1385..1409
                     /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(1409..1428)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1410..1428)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1512..1531)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1737..1814
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1815..2897
                     /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(1834..1853)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3706..3897
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3891..3913)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3999..4141
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4054..4073
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4156..4173)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4327..4915)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4407..4426)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5026..5045)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5037..5852
                     /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             5037..5852
                     /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(5110..5129)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcagatggcg tggttttgct tcattgcagc
       61 aatttcatcg tcgctataac ccagctcctg caatacagca gcggtatgtt cacctaatag
      121 cggcgcagct ttaatatccg gcgtaaaggc agagaatttc attggacaac caacggtcag
      181 atattttcca cgcaacggtt gttccacttc aacaacactg ccactttggc gcaaagaggg
      241 atcaagtgaa atttctttca tacttaaaac cggtgcacaa ggaatatcaa actgagtcaa
      301 ataggccacc gcttcatgtt tatcaatagt gacagtgtat ttttcgattt cagcaaaaat
      361 atcgaaaata tgtggctgtc gtgcatgggc tgtactgtat gccggatcgg taatccattc
      421 tggtttgccg atggctttac aggtgttttc ccagttttgc tcctgaatag tgaaataaat
      481 ataggcgtta ggatcggttt cccagccttt acatttcagg atccagccag gctgaccgcc
      541 accacctgca ttaccaccgc ggggaactgc atcaccaaat gtaccattcg gatactgcgg
      601 gtattcttcc agataaccca atttatcgag acgctgctgg tcacgtaatt tcacgcggca
      661 aaggttcaat acggcatcct gcattgacat ggtgactcgt tgcccacgcc ccgttttttc
      721 gcgatgcagc aaagcagcaa gtaaaccgat cagcaaatgc attccggtgt tgctgtcacc
      781 caacgctgca gcgcttacca gcggcggacc atcccaaaaa cccgtagtgg atgccgcgcc
      841 acccgctgcc tgagcaacgt tttcataggc ttttacattc acataaggcg aacactcatc
      901 aaaccctttg atcgaaccaa aaatcagacg tggattgatt tcttgaatat gctcccaggt
      961 gaagcccatg tgatcaatgg cccctggatg aaagttctcg actaagatat cagcttcgcg
     1021 gatcagcttt tccattacct ctttgccttc cgctgttttg gtatttaact caatagaacg
     1081 tttgttactg ttaagcatgg tgaagtaaag cgcatcgata tcaggaatat ctcgcagctg
     1141 gtgacgcgtt acgtcaccaa cgccgggacg ttcaatttta ataacgtcag cgccaaacca
     1201 ggccagcatt tgagtacaag atgggccaga ttgcacaccg gtgaaatcga gaactttaat
     1261 tccttgaagt ggagttgaca tatggctgcc gcgcggcacc aggccgctgc tgtgatgatg
     1321 atgatgatgg ctgctgccca tggtatatct ccttcttaaa gttaaacaaa attatttcta
     1381 gaggggaatt gttatccgct cacaattccc ctatagtgag tcgtattaat ttcgcgggat
     1441 cgagatctcg atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
     1501 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
     1561 gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt
     1621 gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct
     1681 actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgag atcccggaca
     1741 ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
     1801 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     1861 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     1921 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     1981 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     2041 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     2101 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     2161 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     2221 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     2281 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
     2341 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     2401 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     2461 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     2521 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     2581 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
     2641 ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
     2701 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     2761 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     2821 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     2881 tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcaccg
     2941 ggatctcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     3001 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     3061 gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg
     3121 atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact
     3181 ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggcccca
     3241 cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
     3301 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
     3361 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
     3421 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
     3481 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg
     3541 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt
     3601 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt
     3661 atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa
     3721 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa
     3781 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct
     3841 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     3901 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     3961 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     4021 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
     4081 tgcaccatat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     4141 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4201 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4261 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4321 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4381 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4441 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4501 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4561 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4621 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4681 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4741 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     4801 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4861 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4921 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4981 tttggtcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     5041 gccatattca acgggaaacg tcttgctcta ggccgcgatt aaattccaac atggatgctg
     5101 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     5161 gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     5221 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     5281 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     5341 ccgggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     5401 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     5461 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     5521 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     5581 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     5641 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     5701 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     5761 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     5821 agtttcattt gatgctcgat gagtttttct aagaattaat tcatgagcgg atacatattt
     5881 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5941 cct
//
