LOCUS       sequence-107149-        5175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of O-acetyl-L-homoserine sulfhydrylase (OAH2) of Thermus 
            thermophilus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5175)
  AUTHORS   Poulin MB, Du Q, Schramm VL
  TITLE     Chemoenzymatic Synthesis of (36)S Isotopologues of Methionine and 
            S-Adenosyl-l-methionine.
  JOURNAL   J Org Chem. 2015 May 15;80(10):5344-7. doi: 10.1021/acs.joc.5b00608.
            Epub 2015 Apr 23.
  PUBMED    25884979
REFERENCE   2  (bases 1 to 5175)
  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: 
            "10.1021/acs.joc.5b00608"; journalName: "J Org Chem"; date: 
            "2015-05-15- 15"; volume: "80"; issue: "10"; pages: "5344-7"
COMMENT     Sequence Label: OAH2
FEATURES             Location/Qualifiers
     source          1..5175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      552..1140
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1041..1060
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1294..1311
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1409..1426
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1410..1426
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1890..1909
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1890..1908
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1909..1933
                     /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)."
     terminator      3219..3266
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3489..3508)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        3575..3666
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3667..4476
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             join(4559..5175,1..466)
                     /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(4578..4597)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
       61 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
      121 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
      181 ggatatctcg gtagtgggat acgacgatac cgaagatagc tcatgttata tcccgccgtt
      241 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
      301 actctctcag ggccaggcgg tgaagggcaa tcagctgttg ccagtctcac tggtgaaaag
      361 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
      421 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgactca tgaccaaaat
      481 cccttaacgt gagttacgcg cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
      541 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
      601 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      661 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
      721 gcccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      781 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      841 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      901 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      961 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     1021 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     1081 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     1141 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     1201 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     1261 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     1321 ggcgagagta gggaactgcc aggcatcaaa ctaagcagaa ggcccctgac ggatggcctt
     1381 tttgcgtttc tacaaactct ttctgtgttg taaaacgacg gccagtctta agctcgggcc
     1441 ccctgggcgg ttctgataac gagtaatcgt taatccgcaa ataacgtaaa aacccgcttc
     1501 ggcgggtttt tttatggggg gagtttaggg aaagagcatt tgtcagaata tttaagggcg
     1561 cctgtcactt tgcttgatat atgagaatta tttaacctta taaatgagaa aaaagcaacg
     1621 cactttaaat aagatacgtt gctttttcga ttgatgaaca cctataatta aactattcat
     1681 ctattattta tgattttttg tatatacaat atttctagtt tgttaaagag aattaagaaa
     1741 ataaatctcg aaaataataa agggaaaatc agtttttgat atcaaaatta tacatgtcaa
     1801 cgataataca aaatataata caaactataa gatgttatca gtatttatta tgcatttaga
     1861 ataaattttg tgtcgccctt ccgcgaaatt aatacgactc actatagggg aattgtgagc
     1921 ggataacaat tcccctctag aaataatttt gtttaacttt taggaggtaa aacatatgga
     1981 atatacgacg ttggcagttt tggcaggctt gcctgaagat cctcatggtg cggttggtct
     2041 gccgatttac gcagttgcgg cgtacggttt caaaaccctg gaagagggcc aagagcgttt
     2101 tgcgaccggt gagggttatg tctacgcacg ccaaaaggac ccgaccgcca aggccctgga
     2161 ggagcgtctg aaggcactgg aaggcgcgtt ggaagcggtc gttctggcta gcggtcaggc
     2221 agccaccttt gcggctctgc tggcattgct gcgtccaggc gacgaggtcg tcgccgcgaa
     2281 gggtctgttc ggtcaaacga ttggcctgtt tggtcaggtg ctgagcctga tgggcgtgac
     2341 ggttcgttac gttgatccag agccggaagc ggtgcgcgag gccctgagcg caaaaacccg
     2401 cgcagtgttc gtcgaaaccg tggcgaaccc ggcgttgctg gtcccggatc tggaagccct
     2461 ggcgaccctg gcggaagagg ctggtgtcgc cttagtggtc gacaacacgt ttggcgctgc
     2521 gggtgcgttg tgccgtccgc tggcgtgggg tgcacatgtc gtggttgaat ctctgacgaa
     2581 atgggcttcg ggccacggta gcgtgctggg cggtgcggtt ctctcccgcg agactgagct
     2641 gtggcgtaat tatccgcaat ttctgcagcc ggatctgaag ggtcaaatcc cgtgggaagc
     2701 gctccgtgca cgttgcttcc cggagcgtgt ccgtaccctg ggcctgagcc tgtgtggcat
     2761 ggcgctgagc ccgttcaatg catatctgct gttccagggt ctggaaaccg ttgcgctgcg
     2821 tgttgcgcgt atgagcgaga ctgcacgttt cctggcggag cgcctgcagg gccacccgaa
     2881 agttaaagcg ttgcgttacc cgggtctgcc tgaggacccg gctcatcgta acgcacgcaa
     2941 gtacctggcg tctggtggtc cgatcctgac gctggatctg ggtgatctgg agcgtgcctc
     3001 ccgctttctg ggtgccatcc gcctgctgaa agcggcaaat ctgggcgacg ctcgcaccct
     3061 gcttgtgcac ccgtggacga ccacccacag ccgtctgaaa gaagaagcgc gtctgcaggc
     3121 tggcgtcacg ccgggcctgg tgcgcgtgag cgtgggtttg gaggacccgc tggatttgct
     3181 ggccctgttc gaggaagcat tagaggccgt gtaaccccct agcataaccc cttggggcct
     3241 ctaaacgggt cttgaggggt tttttgcccc tgagacgcgt caatcgagtt cgtacctaag
     3301 ggcgacaccc cctaattagc ccgggcgaaa ggcccagtct ttcgactgag cctttcgttt
     3361 tatttgatgc ctggcagttc cctactctcg catggggagt ccccacacta ccatcggcgc
     3421 tacggcgttt cacttctgag ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc
     3481 caggcaaaca aggggtgtta tgagccatat tcaggtataa atgggctcgc gataatgttc
     3541 agaattggtt aattggttgt aacactgacc cctatttgtt tatttttcta aatacattca
     3601 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3661 aagaatatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     3721 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     3781 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     3841 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcca
     3901 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     3961 atccccggaa aaacagcgtt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     4021 gttgatgcgc tggcagtgtt cctgcgccgg ttgcactcga ttcctgtttg taattgtcct
     4081 tttaacagcg atcgcgtatt tcgcctcgct caggcgcaat cacgaatgaa taacggtttg
     4141 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     4201 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     4261 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     4321 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     4381 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     4441 ttgcagtttc atttgatgct cgatgagttt ttctaagcgg cgcgccatcg aatggcgcaa
     4501 aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatat
     4561 gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc
     4621 ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc
     4681 gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc
     4741 gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc
     4801 ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg
     4861 aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg
     4921 gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac
     4981 taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt
     5041 ctcccatgag gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca
     5101 aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg
     5161 gcataaatat ctcac
//
