LOCUS       sequence-173996-        6536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Irp9 WT, the salicylate synthase from Yersinia 
            enterocolitica.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6536)
  AUTHORS   Meneely KM, Luo Q, Lamb AL
  TITLE     Redesign of MST enzymes to target lyase activity instead promotes 
            mutase and dehydratase activities.
  JOURNAL   Arch Biochem Biophys. 2013 Nov 1;539(1):70-80. doi: 
            10.1016/j.abb.2013.09.007. Epub 2013 Sep 19.
  PUBMED    24055536
REFERENCE   2  (bases 1 to 6536)
  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: 
            "10.1016/j.abb.2013.09.007"; journalName: "Arch Biochem Biophys"; 
            date: "2013-11-1- 1"; volume: "539"; issue: "1"; pages: "70-80"
COMMENT     Sequence Label: pET29b-irp9 WT
FEATURES             Location/Qualifiers
     source          1..6536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     116..135
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     223..242
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        223..241
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    242..266
                     /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)."
     RBS             281..303
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1619..1636
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1689..1707)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1703..1750
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1787..2242
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1874..1893)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2084..2105
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2334..3149)
                     /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             complement(2334..3149)
                     /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     3057..3076
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3141..3160
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3271..3859
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3760..3779
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4013..4030
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4045..4184
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4110..4129)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4270..4292
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4286..4477)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5286..6368)
                     /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     6330..6349
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6369..6446)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
       61 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
      121 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
      181 gtccggcgta gaggatcgag atcgatctcg atcccgcgaa attaatacga ctcactatag
      241 gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg
      301 agatatacat atgaaaatca gtgaatttct acacctggcg ttaccagagg aacaatggct
      361 gccgacgatt tctggcgttt tacgccagtt cgcagaagaa gagtgttatg tctatgagcg
      421 ccaaccctgt tggtatttag gcaaagggtg ccaggcacgg ctgcacatta atgccgacgg
      481 aacgcaggcg acatttattg atgatgccgg ggagcaaaaa tgggcggtgg attccattgc
      541 cgactgcgcg cgtcgtttta tggcacatcc tcaggtgaaa ggacgtcggg tatatggaca
      601 ggttgggttc aactttgcgg cgcatgcgcg ggggattgcc tttaacgccg gggagtggcc
      661 gctgctgacg ttaaccgttc cccgtgaaga acttattttt gaaaagggaa atgtcaccgt
      721 ttatgcggac tccgccgacg ggtgccgacg tttgtgcgag tgggtaaaag aggccagtac
      781 aacgacgcag aacgcaccac tggcggtgga taccgccctc aatggtgagg cgtataaaca
      841 acaggttgcg cgcgccgttg cggagatccg ccgtggcgag tatgtcaaag tgattgtctc
      901 gcgcgccatt cccctgccat cgcggattga tatgcccgcc acgctgttat acgggcgaca
      961 ggcaaacacg ccagtgcgct cgtttatgtt ccgtcaggaa ggacgcgaag cgctgggctt
     1021 tagcccggaa ctggtgatgt cagtgacggg caataaagtg gttactgaac cgcttgcggg
     1081 cacccgcgat cgcatgggaa acccggagca taataaggcg aaagaggcag aactgctgca
     1141 cgatagtaaa gaggtgcttg agcatatcct ttctgtcaaa gaagctattg ctgaactgga
     1201 ggccgtttgc ctgccgggca gcgtggtggt tgaagattta atgtcggttc gccagcgcgg
     1261 cagcgttcag catctggggt ctggcgtgag cggtcagctt gcggaaaaca aggatgcctg
     1321 ggatgcgttt accgtgctgt ttccgtcgat taccgcctca ggtatcccta aaaacgctgc
     1381 cctgaacgcg attatgcaaa ttgagaagac gccgcgagag ctttattccg gcgcaattct
     1441 gctgctggac gatacgcgct tcgatgcggc gctagttctg cgttccgtat ttcaggacag
     1501 ccagcgctgc tggatacagg cgggggcggg aatcatcgcg caatctacac cggaacgcga
     1561 actgacggaa acccgggaga aattagcgag cattgcgccc tatttaatgg tgctcgagca
     1621 ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     1681 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     1741 gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc
     1801 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     1861 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     1921 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     1981 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2041 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2101 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2161 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2221 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2281 tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa
     2341 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     2401 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     2461 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     2521 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     2581 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     2641 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     2701 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     2761 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     2821 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     2881 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     2941 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     3001 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     3061 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag
     3121 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     3181 gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3241 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3301 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3361 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     3421 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3481 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3541 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3601 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3661 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3721 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3781 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3841 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3901 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3961 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4021 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4081 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4141 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     4201 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4261 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4321 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     4381 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     4441 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     4501 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
     4561 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
     4621 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg
     4681 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc
     4741 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg
     4801 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
     4861 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
     4921 ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc
     4981 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga
     5041 ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa
     5101 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     5161 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     5221 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     5281 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5341 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     5401 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     5461 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     5521 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg
     5581 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     5641 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     5701 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     5761 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     5821 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     5881 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     5941 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     6001 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     6061 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     6121 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     6181 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     6241 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     6301 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     6361 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     6421 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     6481 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caagga
//
