LOCUS       sequence-342860-        5013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sMHT-113.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5013)
  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 5013)
  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: pEMF034
FEATURES             Location/Qualifiers
     source          1..5013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             32..43
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             50..763
                     /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"
     terminator      complement(881..908)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(924..995)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        complement(join(1550..1575,1576..1581,1582..1598,1599..1604,
                     1605..1623))
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1580..1598
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1605..1623
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(1644..1662)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             1831..2490
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1897..1916)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2511..2605
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2693..3281
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3182..3201
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3446..3532)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3538..4164)
                     /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     4069..4088
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(4199..4233)
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     promoter        complement(join(4888..4913,4914..4919,4920..4936,4937..4942,
                     4943..4961))
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    4918..4936
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4943..4961
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 tacccgcctc atcagagata atgcttaaga aaaagaggag aaatactaga tgcgtaaagg
       61 cgaagagctg ttcactggtg tcgtccctat tctggtggaa ctggatggtg atgtcaacgg
      121 tcataagttt tccgtgcgtg gcgagggtga aggtgacgca actaatggta aactgacgct
      181 gaagttcatc tgtactactg gtaaactgcc ggtaccttgg ccgactctgg taacgacgct
      241 gacttatggt gttcagtgct ttgctcgtta tccggaccat atgaagcagc atgacttctt
      301 caagtccgcc atgccggaag gctatgtgca ggaacgcacg atttccttta aggatgacgg
      361 cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat accctggtaa accgcattga
      421 gctgaaaggc attgacttta aagaagacgg caatatcctg ggccataagc tggaatacaa
      481 ttttaacagc cacaatgttt acatcaccgc cgataaacaa aaaaatggca ttaaagcgaa
      541 ttttaaaatt cgccacaacg tggaggatgg cagcgtgcag ctggctgatc actaccagca
      601 aaacactcca atcggtgatg gtcctgttct gctgccagac aatcactatc tgagcacgca
      661 aagcgttctg tctaaagatc cgaacgagaa acgcgatcat atggttctgc tggagttcgt
      721 aaccgcagcg ggcatcacgc atggtatgga tgaactgtac aaatgatgac agctcatttc
      781 agaatatttg ccagaaccgt tatgatgtcg gcgcaaaaaa cattatccag aacgggagtg
      841 cgccttgagc gacacgaatt atgcagtgat ttactataaa cgcagaaagg cccacccgaa
      901 ggtgagccag tgtgactcta gtagagagcg ttcaccgaca aacaacagat aaaacgaaag
      961 gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tggtctagat tattatttat
     1021 acgcttcgat ttctttgcgc aggttcgcga ttttctgttt cagctgttcg cgtttctgtt
     1081 tcagctgttc gatgcggttg cgcagttccg cttttttaga tttcagttct tctttttcgt
     1141 tcatggaccc accacctcca gagccaccgc cacccgcggc cgccggcatg gtgctaaagg
     1201 tttcgcgcgc ttttttcagc gcgttttcgc taatatccag gcccaccata aagcgttccg
     1261 ggttcgccat cgccaccaca tcatagccgc cgccgcagcc cggcaccagg cctttgccgt
     1321 tcggcagcgc gttgtttttc accagatgca gcaccagcgg ggtcggcacg cccagatccc
     1381 acggaatcac ttcttccacc cagcattttt cccagccgcc gctgttcacc actttataca
     1441 gaaaggtcgc gcattcttcc ggggtcggaa tggttttgca atcgccggta aacaccggcg
     1501 caatgttcgc cacggtgctc atcaaaactt cctttgtatg ggtcctatcg gtcagtgcgt
     1561 cctgctgatg tgctcagtat ctctatcact gatagggatg tcaatctcta tcactgatag
     1621 ggactcgagg tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     1681 gataataatg gtttcttaga cgtcgatatc tggcgaaaat gagacgttgg ccggcccgta
     1741 agaggttcca actttcacca taatgaaata agatcactac cggtcgtatt ttttgagtta
     1801 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
     1861 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
     1921 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
     1981 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
     2041 ccggaatttc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
     2101 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
     2161 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta tggtgaaaac
     2221 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg
     2281 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
     2341 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
     2401 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
     2461 tactgcgatg agtggcaggg cggggcgtaa tttgatatcg agctcgcttg gactcctgtt
     2521 gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg
     2581 ccgggcgttt tttattggtg agaatccaag cactagtcat gaccaaaatc ccttaacgtg
     2641 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2701 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2761 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2821 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2881 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2941 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3001 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3061 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3121 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3181 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3241 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3301 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttcttgcat gcgttatccc
     3361 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3421 gaacgcccta ggtctagggc ggcggatttg tcctactcag gagagcgttc accgacaaac
     3481 aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgcctta
     3541 agacccactt tcacatttaa gttgtttttc taatccgcat atgatcaatt caaggccgaa
     3601 taagaaggct ggctctgcac cttggtgatc aaataattcg atagcttgtc gtaataatgg
     3661 cggcatacta tcagtagtag gtgtttccct ttcttcttta gcgacttgat gctcttgatc
     3721 ttccaatacg caacctaaag taaaatgccc cactgcgctg agtgcatata atgcattctc
     3781 tagtgaaaaa ccttgttggc ataaaaaggc taattgattt tcgagagttt catactgttt
     3841 ttctgtaggc cgtgtaccta aatgtacttt tgctccatcg cgatgactta gtaaagcaca
     3901 tctaaaactt ttagcgttat tacgtaaaaa atcttgccag ctttcccctt ctaaagggca
     3961 aaagtgagta tggtgcctat ctaacatctc aatggctaag gcgtcgagca aagcccgctt
     4021 attttttaca tgccaataca atgtaggctg ctctacacct agcttctggg cgagtttacg
     4081 ggttgttaaa ccttcgattc cgacctcatt aagcagctct aatgcgctgt taatcacttt
     4141 acttttatct aaacgagaca tcatactacc ccctgttaaa gagtgtttct ctggctgggc
     4201 tagcactata cctaggactg agctagccgt aaaaagcttg catgcctgca ggtcgactct
     4261 agaggatcca cgggtacctt ttgttatcaa taaaaaaggc cccccgattt gggaggcctt
     4321 tttagttaga ttattagttg ttcatttttt tccagcgcgc cagtttttct ttgcctttgc
     4381 gcggttccac cgccagatcg ctataatctt ccaggctcag ctgtttaaag cccagcggca
     4441 ccagcacttt ttcatattcg ctttcgctca cgctaaacgg cgggccgcct tcatggttgg
     4501 taatcggata catcagggta atcagttcgc catccggttt cagcagttca tacatcgctt
     4561 tgccccacgc cgggcgcatt ttcggatcaa tcgcgcaaaa aaacacataa tcaaaaataa
     4621 aatcaaacgg ctgttccggg cgccaggtaa acacatcttc tttcacaaag ctaaagcagc
     4681 tgctgtttgc ggccgcggac ccaccacctc cagagccacc gccaccgaaa taagcttcct
     4741 cacgctctaa ttttgccagg tcacgctcga gattcgcgat atctttctcc agacgagcat
     4801 tttcattctc aaggcgtgct aagtcgtttt ccagggtggc cgcgatagac atttaataac
     4861 ctgctccgtg tgacgttttc gttagttggt cagtgcgtcc tgctgatgtg ctcagtatct
     4921 ctatcactga tagggatgtc aatctctatc actgataggg aaccggtaga gtctcttccc
     4981 ctatatagcc aatcattctt actccctttg aat
//
