LOCUS       sequence-325947-        5016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sMHT-129.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5016)
  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 5016)
  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: pEMF036
FEATURES             Location/Qualifiers
     source          1..5016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(394..488)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(509..1168)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1083..1102
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1337..1355
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1376..1394,1395..1400,1401..1417,1418..1423,
                     1424..1449)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1376..1394
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1401..1419
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      2052..2123
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2139..2166
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2284..2997)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     RBS             3004..3015
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        join(3099..3117,3118..3123,3124..3140,3141..3146,
                     3147..3172)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    3099..3117
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3124..3142
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        3782..3816
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             3851..4477
                     /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(3927..3946)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      4483..4569
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(join(4734..5016,1..306))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4814..4833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
       61 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
      121 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
      181 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
      241 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
      301 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
      361 cgttaaggga ttttggtcat gactagtgct tggattctca ccaataaaaa acgcccggcg
      421 gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa
      481 caggagtcca agcgagctcg atatcaaatt acgccccgcc ctgccactca tcgcagtact
      541 gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct
      601 gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa
      661 cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc
      721 agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt
      781 tttcaccata acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt
      841 ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag
      901 ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacga aattccggat
      961 gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt
     1021 tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt
     1081 gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg
     1141 tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata
     1201 actcaaaaaa tacgaccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     1261 cgggccggcc aacgtctcat tttcgccaga tatcgacgtc taagaaacca ttattatcat
     1321 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcacc tcgagtccct
     1381 atcagtgata gagattgaca tccctatcag tgatagagat actgagcaca tcagcaggac
     1441 gcactgaccg ataggaccca tacaaaggaa gttttgatga gcaccgtggc gaacattgcg
     1501 ccggtgttta ccggcgattg caaaaccatt ccgaccccgg aagaatgcgc gacctttctg
     1561 tataaagtgg tgaacagcgg cggctgggaa aaatgctggg tggaagaagt gattccgtgg
     1621 gatctgggcg tgccgacccc gctggtgctg catctggtga aaaacaacgc gctgccgaac
     1681 ggcaaaggcc tggtgccggg ctgcggcggc ggctatgatg tggtggcgat ggcgaacccg
     1741 gaacgcttta tggtgggcct ggatattagc gaaaacgcgc tgaaaaaagc gcgcgaaacc
     1801 tttagcacca tgccgaacag cagctgcttt agctttgtga aagaagatgt gtttacctgg
     1861 cgcgcggccg cgggtggcgg tggctctgga ggtggtgggt ccatgaacga aaaagaagaa
     1921 ctgaaatcta aaaaagcgga actgcgcaac cgcatcgaac agctgaaaca gaaacgcgaa
     1981 cagctgaaac agaaaatcgc gaacctgcgc aaagaaatcg aagcgtataa ataataatct
     2041 agaccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     2101 gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt
     2161 tctgcgttta tagtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg
     2221 ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctgtca
     2281 tcatttgtac agttcatcca taccatgcgt gatgcccgct gcggttacga actccagcag
     2341 aaccatatga tcgcgtttct cgttcggatc tttagacaga acgctttgcg tgctcagata
     2401 gtgattgtct ggcagcagaa caggaccatc accgattgga gtgttttgct ggtagtgatc
     2461 agccagctgc acgctgccat cctccacgtt gtggcgaatt ttaaaattcg ctttaatgcc
     2521 atttttttgt ttatcggcgg tgatgtaaac attgtggctg ttaaaattgt attccagctt
     2581 atggcccagg atattgccgt cttctttaaa gtcaatgcct ttcagctcaa tgcggtttac
     2641 cagggtatcg ccttcaaatt tcacttccgc acgcgttttg tacgtgccgt catccttaaa
     2701 ggaaatcgtg cgttcctgca catagccttc cggcatggcg gacttgaaga agtcatgctg
     2761 cttcatatgg tccggataac gagcaaagca ctgaacacca taagtcagcg tcgttaccag
     2821 agtcggccaa ggtaccggca gtttaccagt agtacagatg aacttcagcg tcagtttacc
     2881 attagttgcg tcaccttcac cctcgccacg cacggaaaac ttatgaccgt tgacatcacc
     2941 atccagttcc accagaatag ggacgacacc agtgaacagc tcttcgcctt tacgcatcta
     3001 gtatttctcc tctttttctt aagcattatc tctgatgagg cgggtaattc aaagggagta
     3061 agaatgattg gctatatagg ggaagagact ctaccggttc cctatcagtg atagagattg
     3121 acatccctat cagtgataga gatactgagc acatcagcag gacgcactga ccaactaacg
     3181 aaaacgtcac acggagcagg ttattaaatg tctatcgcgg ccaccctgga aaacgactta
     3241 gcacgccttg agaatgaaaa tgctcgtctg gagaaagata tcgcgaatct cgagcgtgac
     3301 ctggcaaaat tagagcgtga ggaagcttat ttcggtggcg gtggctctgg aggtggtggg
     3361 tccgcggccg caccggaaca gccgtttgat tttatttttg attatgtgtt tttttgcgcg
     3421 attgatccga aaatgcgccc ggcgtggggc aaagcgatgt atgaactgct gaaaccggat
     3481 ggcgaactga ttaccctgat gtatccgatt accaaccatg aaggcggccc gccgtttagc
     3541 gtgagcgaaa gcgaatatga aaaagtgctg gtgccgctgg gctttaaaca gctgagcctg
     3601 gaagattata gcgatctggc ggtggaaccg cgcaaaggca aagaaaaact ggcgcgctgg
     3661 aaaaaaatga acaactaata ataatctaac taaaaaggcc tcccaaatcg gggggccttt
     3721 tttattgata acaaaaggta cccgtggatc ctctagagtc gacctgcagg catgcaagct
     3781 ttttacggct agctcagtcc taggtatagt gctagcccag ccagagaaac actctttaac
     3841 agggggtagt atgatgtctc gtttagataa aagtaaagtg attaacagcg cattagagct
     3901 gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt
     3961 agagcagcct acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttagc
     4021 cattgagatg ttagataggc accatactca cttttgccct ttagaagggg aaagctggca
     4081 agatttttta cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg
     4141 agcaaaagta catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca
     4201 attagccttt ttatgccaac aaggtttttc actagagaat gcattatatg cactcagcgc
     4261 agtggggcat tttactttag gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga
     4321 agaaagggaa acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt
     4381 atttgatcac caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg
     4441 attagaaaaa caacttaaat gtgaaagtgg gtcttaaggc atcaaataaa acgaaaggct
     4501 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     4561 aggacaaatc cgccgcccta gacctagggc gttcggctgc ggcgagcggt atcagctcac
     4621 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcatgcaa gaacatgtga
     4681 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4741 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4801 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4861 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4921 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4981 ggctgtgtgc acgaaccccc cgttcagccc gaccgc
//
