LOCUS       sequence-327984-        7402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sMHT-113 fused to CheY only.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7402)
  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 7402)
  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: pDH008
FEATURES             Location/Qualifiers
     source          1..7402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             861..878
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(931..949)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      945..992
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1029..1484
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1116..1135)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1326..1347
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1576..2391)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1576..2391)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2383..2402
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2513..3101
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3002..3021
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3255..3272
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3287..3426
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3352..3371)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3512..3534
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3528..3719)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4528..5610)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5572..5591
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5611..5688)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5894..5913
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5997..6015
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6016..6040
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             6055..6077
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6096..6113
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6123..6140
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             6144..6176
                     /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        6531..6565
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             6583..7395
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="SHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGK
                     PDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKT
                     AFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDAS
                     DFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIA
                     DRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             6583..7395
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="SHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGK
                     PDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKT
                     AFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDAS
                     DFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIA
                     DRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcaa gaaataattt
       61 tgtttaactt taagaaggag atatacatat gggaaagaga gttttgatag tcgatgatgc
      121 agcgttcatg aggatgatgt tgaaagatat catcacaaaa gcgggatacg aagttgcagg
      181 ggaagcaaca aacggtcgtg aagccgttga aaaatacaag gaactcaaac cggacatcgt
      241 cacgatggac atcaccatgc cggagatgaa cggtatcgat gcgatcaaag agatcatgaa
      301 gatcgacccc aacgcaaaga tcatcgtctg cagtgccatg ggacagcagg caatggtcat
      361 agaagctata aaagctggag cgaaagactt cattgtgaaa cccttccagc cttccagggt
      421 ggtagaggct ctcaacaagg tttcgaaggg tgcttctggt ggtggttctt ctggtggtca
      481 catggcggcc gcaaacagca gctgctttag ctttgtgaaa gaagatgtgt ttacctggcg
      541 cccggaacag ccgtttgatt ttatttttga ttatgtgttt ttttgcgcga ttgatccgaa
      601 aatgcgcccg gcgtggggca aagcgatgta tgaactgctg aaaccggatg gcgaactgat
      661 taccctgatg tatccgatta ccaaccatga aggcggcccg ccgtttagcg tgagcgaaag
      721 cgaatatgaa aaagtgctgg tgccgctggg ctttaaacag ctgagcctgg aagattatag
      781 cgatctggcg gtggaaccgc gcaaaggcaa agaaaaactg gcgcgctgga aaaaaatgaa
      841 caactaaaag cttactcgag caccaccacc accaccactg agatccggct gctaacaaag
      901 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
      961 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg
     1021 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1081 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     1141 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     1201 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     1261 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     1321 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     1381 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     1441 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     1501 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1561 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     1621 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     1681 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     1741 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     1801 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     1861 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     1921 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     1981 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     2041 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     2101 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     2161 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     2221 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     2281 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     2341 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct
     2401 tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg
     2461 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2521 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2581 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2641 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2701 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2761 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     2821 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2881 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     2941 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3001 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3061 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3121 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3181 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3241 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     3301 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     3361 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     3421 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3481 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3541 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     3601 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     3661 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     3721 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     3781 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
     3841 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     3901 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     3961 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     4021 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     4081 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     4141 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg
     4201 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     4261 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     4321 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     4381 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     4441 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     4501 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4561 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4621 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     4681 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     4741 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     4801 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     4861 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     4921 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     4981 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     5041 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     5101 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     5161 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     5221 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     5281 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     5341 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     5401 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     5461 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     5521 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     5581 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     5641 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     5701 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     5761 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     5821 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     5881 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     5941 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat
     6001 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt
     6061 taactttaag aaggagatat accatgggca gcagccatca tcatcatcat cacagcagcg
     6121 gcctggtgcc gcgcggcagc catatggcta gcatgactgg tggacagcaa atgggtgcgg
     6181 gatccatgag caccgtggcg aacattgcgc cggtgtttac cggcgattgc aaaaccattc
     6241 cgaccccgga agaatgcgcg acctttctgt ataaagtggt gaacagcggc ggctgggaaa
     6301 aatgctgggt ggaagaagtg attccgtggg atctgggcgt gccgaccccg ctggtgctgc
     6361 atctggtgaa aaacaacgcg ctgccgaacg gcaaaggcct ggtgccgggc tgcggcggcg
     6421 gctatgatgt ggtggcgatg gcgaacccgg aacgctttat ggtgggcctg gatattagcg
     6481 aaaacgcgct gaaaaaagcg cgcgaaacct ttagcaccat gccgtaatga tttacggcta
     6541 gctcagtcat aggtatagtg ctagcaggag gtgtactagt tgagccatat tcaacgggaa
     6601 acgtcttgct ctaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     6661 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc
     6721 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     6781 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     6841 atccgtactc ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc
     6901 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     6961 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     7021 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     7081 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     7141 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     7201 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     7261 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     7321 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     7381 gatgagtttt tctaactgca gt
//
