LOCUS       sequence-326093-        4156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive sMHT-113-SYNZIP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4156)
  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 4156)
  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: pDH012
FEATURES             Location/Qualifiers
     source          1..4156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             155..946
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     terminator      958..1052
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1166..1711
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     1612..1631
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1828..1845
                     /label=L4440
                     /note="L4440 vector, forward primer"
     RBS             2155..2163
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2182..2199
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2209..2226
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2230..2262
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     promoter        2794..2828
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     terminator      2872..2899
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2948..2979)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     protein_bind    3056..3089
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    3273..3291
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             3398..3406
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             3929..3955
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      4036..4107
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4123..4150
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 ggtaccagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg ttgggaagcc
       61 ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca ggggatcaag
      121 atctgatcaa gagacaggat gaggatcgtt tcgtatgagg gaagcggtga tcgccgaagt
      181 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
      241 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
      301 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
      361 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
      421 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
      481 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
      541 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
      601 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
      661 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
      721 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
      781 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
      841 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
      901 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataagagc tcgcttggac
      961 tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg
     1021 gttgccgccg ggcgtttttt attggtgaga atccaagcac tagtaacaac ttatatcgta
     1081 tggggctgac ttcaggtgct acatttgaag agataaattg cactgaaatc tagtaatatt
     1141 ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct
     1201 ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact
     1261 ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc ctttcagttt
     1321 agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc agtggctgct
     1381 gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt accggataag
     1441 gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga gcgaactgcc
     1501 tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca gcggaatgac
     1561 accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag ggggaaacgc
     1621 ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg
     1681 atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc tctcacttcc
     1741 ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta agccatttcc
     1801 gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc
     1861 ccctaggtgg agtctggcct caaatacaaa gtctgcaaag tctcgaaata ttaaaccaga
     1921 gaataaataa ttaaacgcca aaatatttct agcaaaaacc ccagttatta aaccgcctaa
     1981 gtcccccagg aaagggggat ataacagtat agattttgtc agccttcagc ttggctttac
     2041 cgtcagagtt gacatcccta tcagtgatag atataatgtg tggagggccc aagttcactt
     2101 aaaaaggaga tcaacaatga aagcaatttt cgtactgaaa catcttaatc atgctaagga
     2161 ggttttctaa tgggcagcag ccatcatcat catcatcaca gcagcggcct ggtgccgcgc
     2221 ggcagccata tggctagcat gactggtgga cagcaaatgg gtgcgggatc catgagcacc
     2281 gtggcgaaca ttgcgccggt gtttaccggc gattgcaaaa ccattccgac cccggaagaa
     2341 tgcgcgacct ttctgtataa agtggtgaac agcggcggct gggaaaaatg ctgggtggaa
     2401 gaagtgattc cgtgggatct gggcgtgccg accccgctgg tgctgcatct ggtgaaaaac
     2461 aacgcgctgc cgaacggcaa aggcctggtg ccgggctgcg gcggcggcta tgatgtggtg
     2521 gcgatggcga acccggaacg ctttatggtg ggcctggata ttagcgaaaa cgcgctgaaa
     2581 aaagcgcgcg aaacctttag caccatgccg gcggccgcgg gtggcggtgg ctctggaggt
     2641 ggtgggtcca tgaacgaaaa agaagaactg aaatctaaaa aagcggaact gcgcaaccgc
     2701 atcgaacagc tgaaacagaa acgcgaacag ctgaaacaga aaatcgcgaa cctgcgcaaa
     2761 gaaatcgaag cgtataaata atgaggatcc tgatttacgg ctagctcagt cctaggtata
     2821 gtgctagcag gaggtgtaag ctgagttggc tgctgccacc gctgagcaat aagaaggcca
     2881 tcctgacgga tggccttttt gcgtttcgaa caattgaaaa aacctcgcgc cttacctgtt
     2941 gagtaatagt caaaagcctc cggtcggagg cttttgactt tctgcttact gaatttcggt
     3001 ggtgccgtta attaaccggt gggccctcat gataataatg gtttcttaga cgtccgaagt
     3061 tcctattctc tagaaagtat aggaacttcc ctagggaatt caaaagatct taagtaagta
     3121 agagtatacg tatatcggct aataacgtat taaggcgctt cggcgccttt ttttatgggg
     3181 gtattttcat cccaatccac acgcccaacg cacagcaaac accacgtcga ccctatcagc
     3241 tgcgtgcttt ctatgagtcg ttgctgcata actccctatc agtgatagag attgacatcc
     3301 ctatcagtga tagatataat gtgtggaggg cccaagttca cttaaaaagg agatcaacaa
     3361 tgaaagcaat tttcgtactg aaacatctta atcatgctaa ggaggttttc taatgtctat
     3421 cgcggccacc ctggaaaacg acttagcacg ccttgagaat gaaaatgctc gtctggagaa
     3481 agatatcgcg aatctcgagc gtgacctggc aaaattagag cgtgaggaag cttatttcgg
     3541 tggcggtggc tctggaggtg gtgggtccgc ggccgcaaac agcagctgct ttagctttgt
     3601 gaaagaagat gtgtttacct ggcgcccgga acagccgttt gattttattt ttgattatgt
     3661 gtttttttgc gcgattgatc cgaaaatgcg cccggcgtgg ggcaaagcga tgtatgaact
     3721 gctgaaaccg gatggcgaac tgattaccct gatgtatccg attaccaacc atgaaggcgg
     3781 cccgccgttt agcgtgagcg aaagcgaata tgaaaaagtg ctggtgccgc tgggctttaa
     3841 acagctgagc ctggaagatt atagcgatct ggcggtggaa ccgcgcaaag gcaaagaaaa
     3901 actggcgcgc tggaaaaaaa tgaacaacta cccttacgat gtgcctgact acgcgtaatg
     3961 actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
     4021 agcaatacca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     4081 atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg
     4141 cctttctgcg tttata
//
