LOCUS       sequence-429598-        5239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Acrylate biosensor. Medium copy plasmid that expresses CFP 
            proportional to acrylate concentration. Compatible with all other 
            pJKR plasmids..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5239)
  AUTHORS   Rogers JK, Guzman CD, Taylor ND, Raman S, Anderson K, Church GM
  TITLE     Synthetic biosensors for precise gene control and real-time 
            monitoring of metabolites.
  JOURNAL   Nucleic Acids Res. 2015 Jul 7. pii: gkv616.
  PUBMED    26152303
REFERENCE   2  (bases 1 to 5239)
  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: "Nucleic 
            Acids Res. 2015 Jul 7. pii: gkv616."
COMMENT     Sequence Label: pJKR-O-acuR
FEATURES             Location/Qualifiers
     source          1..5239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..636
                     /direction=RIGHT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     472..491
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      842..927
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1020..1047
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             1166..1879
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     HFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2125..2210
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2303..2330
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             2449..3162
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     HFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     RBS             3433..3441
                     /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)"
     terminator      4162..4209
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4317..5132)
                     /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(4317..5132)
                     /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     5040..5059
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5133..5224)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tggtgtcggg aatccgtaaa ggatcttctt gagatccttt tacgatcgtc gtaatctcct
       61 gctctgtaaa cgaaaaaacc gcctggggag gcggtttgat cgaaggttaa gtcagttggg
      121 gaactgctta acctggtaac tggctttagt ggagcgcaga taccaaatac tgtcctttca
      181 gtgtagcctc tgttaggcca ccacttcaag actctcgata tctaaatcca ctaattctca
      241 gttaccaatg gctgctgcca gtggcgtttt gtcgtgtctt tccgggttgg actcaagatg
      301 atagttaccg gataaggcgc agcagtcggg ctgaacgggg ggttcttgca cacagcccag
      361 cttggagcga actgtctaca cggaacggga cgtggtgatt tgggtaaagc ctccaccaca
      421 acacggacgc cgcaggacgg gaacaggaga gcgcaagagg gagccatcag ggggaaacgc
      481 ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg
      541 atgttcgtca ggggggcgga gcctatggaa aaacggcttc gctccggcct tattgtctct
      601 ctgctaagta tcctcctggc atcttctagg acgtttcatg gctgttttgg cggatgagag
      661 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat
      721 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
      781 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
      841 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
      901 ggtgaacgct ctcctgagta ggacaaaycc gcsgggagcg gatttgaacg ttgcgaagca
      961 acggccsggr gggkggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     1021 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actctgctag caagtaaggc
     1081 cgacgcttca caaccgcact tgatttaata gaccataccg tctattattt ctggccatat
     1141 ttaactttaa gaaggagata tacatatgag taaaggagaa gaacttttca ctggagttgt
     1201 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga
     1261 gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa
     1321 actacctgtt ccatggccaa cacttgtcac tactttgact tggggtgttc aatgcttttc
     1381 aagataccca gatcatatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta
     1441 tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac gtgctgaagt
     1501 caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg attttaaaga
     1561 agatggaaac attcttggac acaaattgga atacaactat atttcacaca atgtatacat
     1621 cactgcagac aaacaaaaga atggaatcaa agctcatttc aaaattagac acaacattga
     1681 agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc
     1741 tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa
     1801 cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg
     1861 catggatgaa ctatacaaat aactaatgag tgagctaact tacattaatt gcgttgcgcc
     1921 atggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca
     1981 gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc
     2041 ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg
     2101 cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     2161 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa yccgcsggga
     2221 gcggatttga acgttgcgaa gcaacggccs ggrgggkggc gggcaggacg cccgccataa
     2281 actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta
     2341 caaactctgc tagcaagtaa ggccgacgct tcacaaccgc acttgattta atagaccata
     2401 ccgtctatta tttctggcca tatttaactt taagaaggag atatacatat gagtaaagga
     2461 gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg
     2521 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt
     2581 aaatttattt gcactactgg aaaactacct gttccatggc caacacttgt cactactttg
     2641 acttggggtg ttcaatgctt ttcaagatac ccagatcata tgaaacggca tgactttttc
     2701 aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg
     2761 aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag
     2821 ttaaaaggta ttgattttaa agaagatgga aacattcttg gacacaaatt ggaatacaac
     2881 tatatttcac acaatgtata catcactgca gacaaacaaa agaatggaat caaagctcat
     2941 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa
     3001 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
     3061 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta
     3121 acagctgctg ggattacaca tggcatggat gaactataca aataactaat gagtgagcta
     3181 acttacatta attgcgttgc gccttaatta acggcactcc tcagcaaata taatgaccct
     3241 cttgataacc caagagggca ttttttaatg cccatggcgt ttaccacagc taacaccacg
     3301 tcgtccctat ctgctgccct aggtctatga gtggttgctg gataacttta cgggcatgca
     3361 taaggctcgt aggctatatt cagggagacc acaacggttt ccctctacaa ataattttgt
     3421 ttaactttga aataaggagg taatacaaat gccgctgacc gacaccccgc cgtctgttcc
     3481 gcagaaaccg cgtcgtggtc gtccgcgtgg tgctccggac gcttctctgg ctcaccagtc
     3541 tctgatccgt gctggtctgg aacacctgac cgaaaaaggt tactcttctg ttggtgttga
     3601 cgaaatcctg aaagctgctc gtgttccgaa aggttctttc taccactact tccgtaacaa
     3661 agctgacttc ggtctggctc tgatcgaagc ttacgacacc tacttcgctc gtctcctcga
     3721 ccaggcgttc ctggacggtt cgctggctcc gctggctcgt ctgcgtctgt tcacccgtat
     3781 ggctgaagaa ggtatggctc gtcacggttt ccgtcgtggt tgcctggttg gtaacctggg
     3841 tcaggaaatg ggtgctctgc cggacgactt ccgtgctgct ctgatcggtg ttctggaaac
     3901 ctggcagcgt cgtaccgctc agctgttccg tgaagctcag gcttgcggtg aactgtctgc
     3961 tgaccacgac ccggacgctc tggctgaagc tttctggatc ggttgggaag gtgctatcct
     4021 gcgtgctaaa ctggaactgc gtccggaccc gctgcactct ttcacccgta ccttcggtcg
     4081 tcacttcgtt acccgtaccc atgaataata agcgactaaa aaattgaatg taggaaacca
     4141 acatgccagt tcgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     4201 ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacactgtg cgcggaaccc
     4261 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag
     4321 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     4381 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     4441 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     4501 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     4561 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
     4621 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     4681 tgagcgagac gaaatacgcg gtcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     4741 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     4801 tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca
     4861 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     4921 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     4981 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
     5041 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta
     5101 gagcaagacg tttcccgttg aatatggctc atactcttcc tttttcaata ttattgaagc
     5161 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5221 caaataggca tgcagcgct
//
