LOCUS       sequence-326016-        3410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sMHT-113 F1 only.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3410)
  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 3410)
  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: pEMF047
FEATURES             Location/Qualifiers
     source          1..3410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     517..536
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     771..789
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(810..828,829..834,835..851,852..857,858..883)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    810..828
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    835..853
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1438..1509
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1525..1552
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1610..1644
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             1679..2305
                     /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(1755..1774)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2311..2397
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2562..3150)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2642..2661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3238..3332)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(3353..3410,1..602))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg
       61 tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg
      121 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata
      181 ttctcaataa accctttagg gaaataggcc aggttttcac cataacacgc cacatcttgc
      241 gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac
      301 gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc
      361 tcaccgtctt tcattgccat acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg
      421 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta
      481 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa
      541 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc
      601 attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgac cggtagtgat
      661 cttatttcat tatggtgaaa gttggaacct cttacgggcc ggccaacgtc tcattttcgc
      721 cagatatcga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
      781 tcacgaggcc ctttcgtctt cacctcgagt ccctatcagt gatagagatt gacatcccta
      841 tcagtgatag agatactgag cacatcagca ggacgcactg accgatagga cccatacaaa
      901 ggaagttttg atgagcaccg tggcgaacat tgcgccggtg tttaccggcg attgcaaaac
      961 cattccgacc ccggaagaat gcgcgacctt tctgtataaa gtggtgaaca gcggcggctg
     1021 ggaaaaatgc tgggtggaag aagtgattcc gtgggatctg ggcgtgccga ccccgctggt
     1081 gctgcatctg gtgaaaaaca acgcgctgcc gaacggcaaa ggcctggtgc cgggctgcgg
     1141 cggcggctat gatgtggtgg cgatggcgaa cccggaacgc tttatggtgg gcctggatat
     1201 tagcgaaaac gcgctgaaaa aagcgcgcga aacctttagc accatgccgg cggccgcggg
     1261 tggcggtggc tctggaggtg gtgggtccat gaacgaaaaa gaagaactga aatctaaaaa
     1321 agcggaactg cgcaaccgca tcgaacagct gaaacagaaa cgcgaacagc tgaaacagaa
     1381 aatcgcgaac ctgcgcaaag aaatcgaagc gtataaataa taatctagac caggcatcaa
     1441 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     1501 aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatagt
     1561 aaatcactgc ataattcgtg tcgctcaagg cgcactcccg ttcaagcttt ttacggctag
     1621 ctcagtccta ggtatagtgc tagcccagcc agagaaacac tctttaacag ggggtagtat
     1681 gatgtctcgt ttagataaaa gtaaagtgat taacagcgca ttagagctgc ttaatgaggt
     1741 cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag agcagcctac
     1801 attgtattgg catgtaaaaa ataagcgggc tttgctcgac gccttagcca ttgagatgtt
     1861 agataggcac catactcact tttgcccttt agaaggggaa agctggcaag attttttacg
     1921 taataacgct aaaagtttta gatgtgcttt actaagtcat cgcgatggag caaaagtaca
     1981 tttaggtaca cggcctacag aaaaacagta tgaaactctc gaaaatcaat tagccttttt
     2041 atgccaacaa ggtttttcac tagagaatgc attatatgca ctcagcgcag tggggcattt
     2101 tactttaggt tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag aaagggaaac
     2161 acctactact gatagtatgc cgccattatt acgacaagct atcgaattat ttgatcacca
     2221 aggtgcagag ccagccttct tattcggcct tgaattgatc atatgcggat tagaaaaaca
     2281 acttaaatgt gaaagtgggt cttaaggcat caaataaaac gaaaggctca gtcgaaagac
     2341 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     2401 ccgccctaga cctagggcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2461 atacggttat ccacagaatc aggggataac gcatgcaaga acatgtgagc aaaaggccag
     2521 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2581 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2641 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2701 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2761 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2821 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2881 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2941 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3001 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3061 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3121 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3181 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgactag tgcttggatt
     3241 ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc
     3301 tgaggtcatt actggatcta tcaacaggag tccaagcgag ctcgatatca aattacgccc
     3361 cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg
//
