LOCUS       sequence-119410-        5516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Macrolide biosensor. Medium copy plasmid that expresses GFP 
            proportional to macrolide concentration. Compatible with all other 
            pJKR plasmids..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5516)
  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 5516)
  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-mphR
FEATURES             Location/Qualifiers
     source          1..5516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(118..135)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(252..797)
                     /direction=LEFT
                     /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     complement(332..351)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1035..1826)
                     /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"
     primer_bind     complement(2221..2238)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2391..2477
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2569..2596
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             2713..3423
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2858..2876)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3126..3145
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     RBS             3691..3699
                     /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      complement(4306..4400)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        complement(5173..5277)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5345..5363
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      5431..5478
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tcagatcctt ccgtatttag ccagtatgtt ctctagacgg atcgcttcat gtggcaggag
       61 aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc
      121 gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg
      181 gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg
      241 gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca
      301 aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
      361 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
      421 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
      481 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
      541 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
      601 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
      661 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
      721 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
      781 gaccaaaacg atctcaagaa gatcatctta ttaaggggtc tgacgctcag tggaacgaaa
      841 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccagc
      901 ggtatcatca acaggcttac ccgtcttact gtcaagagga catccggtca aataaaacga
      961 aaggctcagt cgaaagactg ggcctttcgt tttgctgagg agacttaggg acccttcgaa
     1021 ccccagagtc ccgcttattt gccgactacc ttggtgatct cgcctttcac gtagtggaca
     1081 aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc
     1141 tgtctagctt caagtatgac gggctgatat tgggccggca ggcgctccat tgcccagtcg
     1201 gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac
     1261 gtaagcacta catttcgctc atcgccagcc cagtcgggcg gcgagttcca tagcgttaag
     1321 gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc
     1381 gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca
     1441 atgtcgatcg tggctggctc gaagatacct gcaagaatgt cattgcgctg ccattctcca
     1501 aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cacaacaatg
     1561 gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt ttccaaaagg
     1621 tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa
     1681 tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa atgtacggcc
     1741 agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag ttgagtcgat
     1801 acttcggcga tcaccgcttc cctcatgatg tttaactttg ttttagggcg actgccctgc
     1861 tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc
     1921 tgcttggatg cccgaggcat agactgtacc ccaaaaaaac agtcataaca agccatgaaa
     1981 accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag
     2041 cgcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct gcgccatcag
     2101 atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag
     2161 ggcgccccag ctggcaattc cgacgtctgg ctgttttggc ggatgagaga agattttcag
     2221 cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg
     2281 cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc
     2341 cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac
     2401 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     2461 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag
     2521 ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc
     2581 tgacggatgg cctttttgcg tttctacaaa cttgctagca agtaaggccg acggattgaa
     2641 tataaccgac gtgactgtta catttaggtg gctaaacccg tcaacattta actttaagaa
     2701 ggagatatac atatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     2761 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     2821 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     2881 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     2941 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     3001 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     3061 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     3121 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     3181 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     3241 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     3301 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     3361 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     3421 taactaatga gtgagctaac ttacattaat tgcgttgcgc cttaattaac ggcactcctc
     3481 agcaaatata atgaccctct tgataaccca agagggcatt ttttaatgcc catggcgttt
     3541 accacagcta acaccacgtc gtccctatct gctgccctag gtctatgagt ggttgctgga
     3601 taactttacg ggcatgcata aggctcgtag gctatattca gggagaccac aacggtttcc
     3661 ctctacaaat aattttgttt aactttgaaa taaggaggta atacaaatgc cgcgtccgaa
     3721 actgaaatct gacgacgaag ttctggaagc ggcgaccgtt gttctgaaac gttgcggtcc
     3781 gatcgaattc accctgtctg gtgttgcgaa agaagttggt ctgtctcgtg cggcgctgat
     3841 ccagcgtttc accaaccgtg acaccctgct ggttcgtatg atggaacgtg gtgttgaaca
     3901 ggttcgtcac tacctgaacg cgatcccgat cggtgcgggt ccgcagggtc tgtgggaatt
     3961 cctgcaggtt ctggttcgtt ctatgaacac ccgtaacgac ttctctgtta actacctgat
     4021 ctcttggtac gaactgcagg ttccggaact gcgtaccctg gcgatccagc gtaaccgtgc
     4081 ggttgttgaa ggtatccgta aacgtctgcc gccgggtgcg ccggcggcgg cggaactgct
     4141 gctgcactct gttatcgcgg gtgcgaccat gcagtgggcg gttgacccgg acggtgaact
     4201 ggcggaccac gttctggcgc agatcgcggc gatcctgtgc ctgatgttcc cggaacacga
     4261 cgacttccag ctgctgcagc cgcacgcgta ataactagtg cttggattct caccaataaa
     4321 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
     4381 tggatctatc aacaggagtc caagcgagct cgtaaacttg gtctgacagc ccttaactta
     4441 cttattaaat aatttatagc tattgaaaag agataagaat tgttcaaagc taatattgtt
     4501 taaatcgtca attcctgcat gttttaagga attgttaaat tgattttttg taaatatttt
     4561 cttgtattct ttgttaaccc atttcataac gaaataatta tacttttgtt tatctttgtg
     4621 tgatattctt gatttttttc tacttaatct gataagtgag ctattcactt taggtttagg
     4681 atgaaaatat tctcttggaa ccatacttaa tatagaaata tcaacttctg ccattaaaag
     4741 taatgccaat gagcgttttg tatttaataa tcttttagca aacccgtatt ccacgattaa
     4801 ataaatctca ttagctatac tatcaaaaac aattttgcgt attatatccg tacttatgtt
     4861 ataaggtata ttaccatata ttttatagga ttggttttta ggaaatttaa actgcaatat
     4921 atccttgttt aaaacttgga aattatcgtg atcaacaagt ttattttctg tagttttgca
     4981 taatttatgg tctatttcaa tggcagttac gaaattacac ctctttacta attcaagggt
     5041 aaaatggcct tttcctgagc cgatttcaaa gatattatca tgttcattta atcttatatt
     5101 tgtcattatt ttatctatat tatgttttga agtaataaag ttttgactgt gttttatatt
     5161 tttctcgttc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5221 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5281 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5341 taaaaatagg cgtatcacga ggccctttcg tcttcactaa gcgactaaaa aattgaatgt
     5401 aggaaaccaa catgccagtt cgagcaataa ctagcataac cccttggggc ctctaaacgg
     5461 gtcttgaggg gttttttgct gaaacctcag gcatttgaga agcacacggt cacact
//
