LOCUS       sequence-318874-        7407 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 7407)
  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 7407)
  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..7407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(54..72)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      68..115
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      152..607
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(239..258)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     449..470
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(700..1515)
                     /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(700..1515)
                     /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     1507..1526
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1637..2225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2126..2145
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2379..2396
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2411..2553
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2479..2498)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2639..2661
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2655..2846)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3655..4737)
                     /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     4699..4718
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4738..4815)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5021..5040
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5124..5142
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5143..5167
                     /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             5182..5204
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5223..5240
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5250..5267
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5271..5303
                     /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        5658..5692
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             5708..6523
                     /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             5708..6523
                     /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"
     promoter        6531..6549
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6550..6574
                     /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             6589..6611
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(7391..7407,1)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
       61 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
      121 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
      181 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
      241 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
      301 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
      361 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
      421 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
      481 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
      541 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
      601 acaatttcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
      661 taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact catcgagcat
      721 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
      781 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
      841 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
      901 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
      961 aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     1021 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     1081 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     1141 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     1201 tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     1261 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     1321 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     1381 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     1441 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg
     1501 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     1561 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1621 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1681 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1741 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     1801 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1861 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1921 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1981 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2041 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2101 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2161 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2221 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2281 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     2341 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     2401 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2461 tatatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     2521 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     2581 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     2641 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     2701 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     2761 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     2821 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     2881 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     2941 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     3001 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     3061 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     3121 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     3181 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     3241 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     3301 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     3361 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     3421 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     3481 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     3541 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     3601 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     3661 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3721 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     3781 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     3841 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     3901 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     3961 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     4021 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     4081 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     4141 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     4201 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     4261 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     4321 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     4381 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     4441 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     4501 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     4561 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     4621 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     4681 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     4741 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     4801 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
     4861 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     4921 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     4981 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     5041 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     5101 tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
     5161 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc atgggcagca
     5221 gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat atggctagca
     5281 tgactggtgg acagcaaatg ggtgcgggat ccatgagcac cgtggcgaac attgcgccgg
     5341 tgtttaccgg cgattgcaaa accattccga ccccggaaga atgcgcgacc tttctgtata
     5401 aagtggtgaa cagcggcggc tgggaaaaat gctgggtgga agaagtgatt ccgtgggatc
     5461 tgggcgtgcc gaccccgctg gtgctgcatc tggtgaaaaa caacgcgctg ccgaacggca
     5521 aaggcctggt gccgggctgc ggcggcggct atgatgtggt ggcgatggcg aacccggaac
     5581 gctttatggt gggcctggat attagcgaaa acgcgctgaa aaaagcgcgc gaaaccttta
     5641 gcaccatgcc gtaatgattt acggctagct cagtcctagg tatagtgcta gcaggaggtg
     5701 tactagtatg agccatattc aacgggaaac gtcttgctct aggccgcgat taaattccaa
     5761 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     5821 gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     5881 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     5941 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     6001 cactgcgatc cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     6061 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     6121 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     6181 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     6241 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     6301 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     6361 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     6421 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     6481 gaataaattg cagtttcatt tgatgctcga tgagtttttc taactgcagt taatacgact
     6541 cactataggg gaattgtgag cggataacaa ttcccctcaa gaaataattt tgtttaactt
     6601 taagaaggag atatacatat gggaaagaga gttttgatag tcgatgatgc agcgttcatg
     6661 aggatgatgt tgaaagatat catcacaaaa gcgggatacg aagttgcagg ggaagcaaca
     6721 aacggtcgtg aagccgttga aaaatacaag gaactcaaac cggacatcgt cacgatggac
     6781 atcaccatgc cggagatgaa cggtatcgat gcgatcaaag agatcatgaa gatcgacccc
     6841 aacgcaaaga tcatcgtctg cagtgccatg ggacagcagg caatggtcat agaagctata
     6901 aaagctggag cgaaagactt cattgtgaaa cccttccagc cttccagggt ggtagaggct
     6961 ctcaacaagg tttcgaaggg tgcttctggt ggtggttctt ctggtggtca catggcggcc
     7021 gcaaacagca gctgctttag ctttgtgaaa gaagatgtgt ttacctggcg cccggaacag
     7081 ccgtttgatt ttatttttga ttatgtgttt ttttgcgcga ttgatccgaa aatgcgcccg
     7141 gcgtggggca aagcgatgta tgaactgctg aaaccggatg gcgaactgat taccctgatg
     7201 tatccgatta ccaaccatga aggcggcccg ccgtttagcg tgagcgaaag cgaatatgaa
     7261 aaagtgctgg tgccgctggg ctttaaacag ctgagcctgg aagattatag cgatctggcg
     7321 gtggaaccgc gcaaaggcaa agaaaaactg gcgcgctgga aaaaaatgaa caactaaaag
     7381 cttactcgag caccaccacc accacca
//
