LOCUS       sequence-341219-        6977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ratiometric gas biosensor plasmid for 3-oxo-C12-HSL sensing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6977)
  AUTHORS   Cheng HY, Masiello CA, Del Valle I, Gao X, Bennett GN, Silberg JJ
  TITLE     Ratiometric gas reporting: a non-disruptive approach to monitor gene
            expression in soils.
  JOURNAL   ACS Synth Biol. 2018 Jan 24. doi: 10.1021/acssynbio.7b00405.
  PUBMED    29366321
REFERENCE   2  (bases 1 to 6977)
  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"; journalName: "ACS Synth 
            Biol. 2018 Jan 24. doi: 10.1021/acssynbio.7b00405."
COMMENT     Sequence Label: pHC01
FEATURES             Location/Qualifiers
     source          1..6977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(733..835)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     RBS             1207..1218
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2146..2164
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2271..2279
                     /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)"
     RBS             3345..3356
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3357..4034
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      4067..4138
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4154..4181
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             4534..4725
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLACFGDDGENL"
     primer_bind     complement(4719..4741)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4827..4967
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4882..4901
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4982..4999)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5153..5741)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5233..5252)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6009..6824)
                     /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="MSHIQRETSCSRPRLNSSMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6732..6751
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6816..6835
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gcttgcgagc tctcagagag taataagacc caaattaacg gccataatgg ccgctacgcg
       61 gcgggaggtc acaccgaact tccgccgaat atttcccata tggaagttca cattggcttc
      121 cgagcagttg cagataaccg atatctccca actggtcttg ccgatggcgc accactgcaa
      181 cacttccttc tcccggctgg tcagaaccac cggtttgctg accggatgtt cgaaggccag
      241 tccggcaccg ctttgcagtg cgtagtcctt gagcatccac agggtcggca ggaccgactc
      301 tatgaaacgg ttggcctcgg cccggttttc cgcttccacg ctgaggctca gcgcgccgag
      361 ttcgccgcga gcaccatgca gcggcatggt cagcccatac accaggtcgg cggccgaggc
      421 ttcctcgaag aactcgtgct gctttcgcgt ctggtagatg gacggttccc agaaaatcgg
      481 cagtgcgctc tgggtacagt gactgaccgt cgggtcgacc cgcgcgtagc cagcccggtc
      541 gtaatgctcg cgccaggcgg ccgggtagtt gccgacgatg aaggcgttct cgtagtcctg
      601 gctgtcctta ggcaacaggc cgaacaggat cttcgagaat ccaaggtcgc tcgccatctt
      661 ctggaggatg gcgctccact ccaattttcc acttgagcgt tccagctcaa gaaaaccgtc
      721 aaccaaggcc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
      781 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaacggc
      841 cacctcgacc tgaatggaag ccggcggcac ctcgctaacg gattcaccac tccaagaatt
      901 ggagccaatc aattcttgcg gagaactgtg aatgcgcaaa ccaacccttg gcagaacata
      961 tccatcgcgt ccgccatctc cagcagccgc acgcggcgca tctcgggcag cgttgggtcc
     1021 tggccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt
     1081 tgccttactg gttagcagaa tgaactagtt tcgagcctag caagggtccg ggttcaccga
     1141 aatctatctc atttgctagt tataaaatta tgaaatttgc gtaaattctt cactcgagta
     1201 ctagagaaag aggagaaata ctagatgagc accgtggcga acattgcgcc ggtgtttacc
     1261 ggcgattgca aaaccattcc gaccccggaa gaatgcgcga cctttctgta taaagtggtg
     1321 aacagcggcg gctgggaaaa atgctgggtg gaagaagtga ttccgtggga tctgggcgtg
     1381 ccgaccccgc tggtgctgca tctggtgaaa aacaacgcgc tgccgaacgg caaaggcctg
     1441 gtgccgggct gcggcggcgg ctatgatgtg gtggcgatgg cgaacccgga acgctttatg
     1501 gtgggcctgg atattagcga aaacgcgctg aaaaaagcgc gcgaaacctt tagcaccatg
     1561 ccgaacagca gctgctttag ctttgtgaaa gaagatgtgt ttacctggcg cccggaacag
     1621 ccgtttgatt ttatttttga ttatgtgttt ttttgcgcga ttgatccgaa aatgcgcccg
     1681 gcgtggggca aagtgatgta tgaactgctg aaaccggatg gcgaactgat taccctgatg
     1741 tatccgatta ccaaccatga aggcggcccg ccgtttagcg tgagcgaaag cgaatatgaa
     1801 aaagtgctgg tgccgctggg ctttaaacag ctgagcctgg aagattatag cgatctggcg
     1861 gtggaaccgc gcaaaggcaa agaaaaactg gcgcgctgga aaaaaatgaa caactaataa
     1921 atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtcga attcaaaaga
     1981 tcttaagtaa gtaagagtat acgtatatcg gctaataacg tattaaggcg cttcggcgcc
     2041 tttttttatg ggggtatttt catcccaatc cacacgtcca acgcacagca aacaccacgt
     2101 cgaccctatc agctgcgtgc tttctatgag tcgttgctgc ataactccct atcagtgata
     2161 gagattgaca tccctatcag tgatagatat aatgtgtgga gggcccaagt tcacttaaaa
     2221 aggagatcaa caatgaaagc aattttcgta ctgaaacatc ttaatcatgc taaggaggtt
     2281 ttctaatgac caatctccag acattcgaat tacctaccga agtcaccgga tgtgccgccg
     2341 atattagcct gggtcgcgca ctgattcagg cgtggcaaaa agacggtatt tttcagatta
     2401 aaaccgattc agaacaggat cgtaaaacac aggaagcgat ggccgccagt aaacagtttt
     2461 gtaaggaacc cctgaccttt aaatcttcat gtgtttcgga tctgacgtat agcggatacg
     2521 tggcaagcgg ggaagaggtc accgcaggca aaccagactt cccggaaatt tttactgtat
     2581 gcaaagacct ctctgtcgga gaccagcgtg tgaaggcggg ttggccatgc catggtccag
     2641 ttccatggcc gaataacact taccaaaaat cgatgaaaac tttcatggaa gaactgggtc
     2701 tggcgggtga gcgcctcctt aagttgacag cactgggctt cgagctccct atcaacacgt
     2761 tcaccgatct gacccgtgat ggttggcacc atatgcgtgt gctgcgcttt ccgccgcaga
     2821 cctcaaccct tagccgcggc atcggggctc atactgatta cggtttactg gtgattgcgg
     2881 cccaggatga cgttggcggt ctttatattc gtccgccggt tgagggcgaa aaacgtaacc
     2941 gtaactggct cccgggcgag agttcggcgg gtatgttcga gcacgacgag ccgtggactt
     3001 tcgtaacgcc gacgccggga gtgtggaccg tttttccggg tgatatctta caatttatga
     3061 ccggcgggca gctcctttca acgcctcata aagtaaagct caacacccgc gaacgttttg
     3121 cttgcgcata ctttcatgaa ccgaacttcg aagccagcgc atacccactg tttgaaccgt
     3181 cggcgaatga acgcattcac tacggagaac acttcaccaa tatgttcatg cgttgttacc
     3241 cggatcgtat tacaacgcag cgcattaata aagaaaaccg cctggcccat ttggaggacc
     3301 tgaaaaaata tagtgacact cgtgccacag gctcgtaata aattaaagag gagaaaatgg
     3361 cgagtagcga agacgttatc aaagagttca tgcgtttcaa agttcgtatg gaaggttccg
     3421 ttaacggtca cgagttcgaa atcgaaggtg aaggtgaagg tcgtccgtac gaaggtaccc
     3481 agaccgctaa actgaaagtt accaaaggtg gtccgctgcc gttcgcttgg gacatcctgt
     3541 ccccgcagtt ccagtacggt tccaaagctt acgttaaaca cccggctgac atcccggact
     3601 acctgaaact gtccttcccg gaaggtttca aatgggaacg tgttatgaac ttcgaagacg
     3661 gtggtgttgt taccgttacc caggactcct ccctgcaaga cggtgagttc atctacaaag
     3721 ttaaactgcg tggtaccaac ttcccgtccg acggtccggt tatgcagaaa aaaaccatgg
     3781 gttgggaagc ttccaccgaa cgtatgtacc cggaagacgg tgctctgaaa ggtgaaatca
     3841 aaatgcgtct gaaactgaaa gacggtggtc actacgacgc tgaagttaaa accacctaca
     3901 tggctaaaaa accggttcag ctgccgggtg cttacaaaac cgacatcaaa ctggacatca
     3961 cctcccacaa cgaagactac accatcgttg aacagtacga acgtgctgaa ggtcgtcact
     4021 ccaccggtgc ttaaggatcg gttgtcgagt aaggatctcc aggcatcaaa taaaacgaaa
     4081 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta
     4141 ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatagga tcctgcgacc
     4201 tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag
     4261 tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt
     4321 ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg
     4381 tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt
     4441 tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc
     4501 atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt
     4561 aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg
     4621 gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac
     4681 cgcagctgcc tcgcgtgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     4741 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     4801 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     4861 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     4921 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     4981 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5041 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5101 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5161 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5221 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5281 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5341 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5401 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatctggt aactatcgtc
     5461 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5521 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5581 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5641 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5701 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgacctttt
     5761 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5821 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5881 aaagtatata tgagtaaact tggtctgaca gttacgcccc gcccggtacc cggggaaatg
     5941 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6001 attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     6061 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     6121 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     6181 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     6241 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
     6301 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     6361 cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca
     6421 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     6481 tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac
     6541 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     6601 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     6661 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     6721 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gctggaattt
     6781 aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
     6841 ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac gtgagttttc
     6901 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6961 tctgcgcgta atctgct
//
