LOCUS       sequence-326018-        3338 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sMHT-113 F2 only.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3338)
  AUTHORS   Fulk EM, Huh D, Atkinson JT, Lie M, Masiello CA, Silberg JJ
  TITLE     A Split Methyl Halide Transferase AND Gate That Reports by 
            Synthesizing an Indicator Gas.
  JOURNAL   ACS Synth Biol. 2020 Nov 20;9(11):3104-3113. doi: 
            10.1021/acssynbio.0c00355. Epub 2020 Oct 26.
  PUBMED    33104325
REFERENCE   2  (bases 1 to 3338)
  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/acssynbio.0c00355"; journalName: "ACS Synth Biol"; date: 
            "2020-11-20- 20"; volume: "9"; issue: "11"; pages: "3104-3113"
COMMENT     Sequence Label: pEMF044
FEATURES             Location/Qualifiers
     source          1..3338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(264..358)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(379..1038)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     953..972
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1207..1225
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1246..1264,1265..1270,1271..1287,1288..1293,
                     1294..1319)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1246..1264
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1271..1289
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        1974..2008
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2043..2669
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(2119..2138)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2675..2761
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(join(2926..3338,1..176))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3006..3025)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
       61 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
      121 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
      181 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
      241 ttttggtcat gactagtgct tggattctca ccaataaaaa acgcccggcg gcaaccgagc
      301 gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca
      361 agcgagctcg atatcaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc
      421 attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag
      481 cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa
      541 gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc
      601 tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt tttcaccata
      661 acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact
      721 ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact
      781 atcccatatc accagctcac cgtctttcat tgccatacga aattccggat gagcattcat
      841 caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt
      901 ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga
      961 ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc
     1021 agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     1081 tacgaccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgggccggcc
     1141 aacgtctcat tttcgccaga tatcgacgtc taagaaacca ttattatcat gacattaacc
     1201 tataaaaata ggcgtatcac gaggcccttt cgtcttcacc tcgagtccct atcagtgata
     1261 gagattgaca tccctatcag tgatagagat actgagcaca tcagcaggac gcactgacca
     1321 actaacgaaa acgtcacacg gagcaggtta ttaaatgtct atcgcggcca ccctggaaaa
     1381 cgacttagca cgccttgaga atgaaaatgc tcgtctggag aaagatatcg cgaatctcga
     1441 gcgtgacctg gcaaaattag agcgtgagga agcttatttc ggtggcggtg gctctggagg
     1501 tggtgggtcc gcggccgcaa acagcagctg ctttagcttt gtgaaagaag atgtgtttac
     1561 ctggcgcccg gaacagccgt ttgattttat ttttgattat gtgttttttt gcgcgattga
     1621 tccgaaaatg cgcccggcgt ggggcaaagc gatgtatgaa ctgctgaaac cggatggcga
     1681 actgattacc ctgatgtatc cgattaccaa ccatgaaggc ggcccgccgt ttagcgtgag
     1741 cgaaagcgaa tatgaaaaag tgctggtgcc gctgggcttt aaacagctga gcctggaaga
     1801 ttatagcgat ctggcggtgg aaccgcgcaa aggcaaagaa aaactggcgc gctggaaaaa
     1861 aatgaacaac taataatcta actaaaaagg cctcccaaat cggggggcct tttttattga
     1921 taacaaaagg tacccgtgga tcctctagag tcgacctgca ggcatgcaag ctttttacgg
     1981 ctagctcagt cctaggtata gtgctagccc agccagagaa acactcttta acagggggta
     2041 gtatgatgtc tcgtttagat aaaagtaaag tgattaacag cgcattagag ctgcttaatg
     2101 aggtcggaat cgaaggttta acaacccgta aactcgccca gaagctaggt gtagagcagc
     2161 ctacattgta ttggcatgta aaaaataagc gggctttgct cgacgcctta gccattgaga
     2221 tgttagatag gcaccatact cacttttgcc ctttagaagg ggaaagctgg caagattttt
     2281 tacgtaataa cgctaaaagt tttagatgtg ctttactaag tcatcgcgat ggagcaaaag
     2341 tacatttagg tacacggcct acagaaaaac agtatgaaac tctcgaaaat caattagcct
     2401 ttttatgcca acaaggtttt tcactagaga atgcattata tgcactcagc gcagtggggc
     2461 attttacttt aggttgcgta ttggaagatc aagagcatca agtcgctaaa gaagaaaggg
     2521 aaacacctac tactgatagt atgccgccat tattacgaca agctatcgaa ttatttgatc
     2581 accaaggtgc agagccagcc ttcttattcg gccttgaatt gatcatatgc ggattagaaa
     2641 aacaacttaa atgtgaaagt gggtcttaag gcatcaaata aaacgaaagg ctcagtcgaa
     2701 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     2761 tccgccgccc tagacctagg gcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2821 ggtaatacgg ttatccacag aatcagggga taacgcatgc aagaacatgt gagcaaaagg
     2881 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2941 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3001 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3061 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3121 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3181 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3241 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3301 agcgaggtat gtaggcggtg ctacagagtt cttgaagt
//
