LOCUS       sequence-102831-        5769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Macrolide biosensor. Low copy plasmid that expresses GFP 
            proportional to macrolide concentration..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5769)
  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 5769)
  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-L-mphR
FEATURES             Location/Qualifiers
     source          1..5769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(47..115,116..907)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(265..284)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(1060..2010)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2058..2280
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     complement(2772..2789)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2942..3028
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             3265..3978
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     RBS             4271..4279
                     /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(4886..4980)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
       61 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      121 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      181 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      241 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      301 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      361 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      421 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
      481 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
      541 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
      601 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
      661 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
      721 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
      781 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
      841 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
      901 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
      961 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1021 acgtgagttt tcgttccact gagcgtcaga ccccgaacct cagatccttc cgtatttagc
     1081 cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg aaccattgag
     1141 atcatactta ctttgcatgt cactcaaaaa ttttgcctca aaactggtga gctgaatttt
     1201 tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta tgtaggtagg aatctgatgt
     1261 aatggttgtt ggtattttgt caccattcat ttttatctgg ttgttctcaa gttcggttac
     1321 gagatccatt tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg
     1381 cttatcaacc accaatttca tattgctgta agtgtttaaa tctttactta ttggtttcaa
     1441 aacccattgg ttaagccttt taaactcatg gtagttattt tcaagcatta acatgaactt
     1501 aaattcatca aggctaatct ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt
     1561 taataaccac tcataaatcc tcatagagta tttgttttca aaagacttaa catgttccag
     1621 attatatttt atgaattttt ttaactggaa aagataaggc aatatctctt cactaaaaac
     1681 taattctaat ttttcgcttg agaacttggc atagtttgtc cactggaaaa tctcaaagcc
     1741 tttaaccaaa ggattcctga tttccacagt tctcgtcatc agctctctgg ttgctttagc
     1801 taatacacca taagcatttt ccctactgat gttcatcatc tgagcgtatt ggttataagt
     1861 gaacgatacc gtccgttctt tccttgtggg gttttcaatc gtggggttga gtagtgccac
     1921 acagcataaa attagcttgg tttcatgctc cgttaagtca tagcgactaa tcgctagttc
     1981 atttgctttg aaaacaacta attcagacat acatctcaat tggtctaggt gattttaatc
     2041 actataccaa ttgagatggg ctagtcaatg ataattacta gtccttttcc tttgagttgt
     2101 gggtatctgt aaattctgct agacctttgc tggaaaactt gtaaattctg ctagaccctc
     2161 tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat attcaagtgg ttataattta
     2221 tagaataaag aaagaataaa aaaagataaa aagaatagat cccagccctg tgtataactc
     2281 actactttag tcagttccgc agtattacaa aaggatgtcg caaacgctgt ttgctcctct
     2341 acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct cgggcaaatc
     2401 gctgaatatt ccttttgtct ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca
     2461 ttcagttcgc tgcgctcacg gctctggcag tgaatggggg taaatggcac tacaggcgcc
     2521 ttttatggat tcatgcaagg aaactaccca taatacaaga aaagcccgtc acgggcttct
     2581 cagggcgttt tatggcgggt ctgctatgtg gtgctatctg actttttgct gttcagcagt
     2641 tcctgccctc tgattttcca gtctgaccac ttcggattat cccgtgacag gtcattcaga
     2701 ctggctaatg cacccagtaa ggcagcggta tcatcaactg gctgttttgg cggatgagag
     2761 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat
     2821 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
     2881 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     2941 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     3001 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     3061 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     3121 gaaggccatc cgtcaggatg gcctttttgc gtttctacaa actctgctag caagtaaggc
     3181 cgacggattg aatataaccg acgtgactgt tacatttagg tggctaaacc cgtcaacatt
     3241 taactttaag aaggagatat acatatgcgt aaaggtgaag aactgttcac cggtgttgtt
     3301 ccgatcctgg ttgaactgga cggtgacgtt aacggtcaca aattctctgt tcgtggtgaa
     3361 ggtgaaggtg acgctaccaa cggtaaactg accctgaaat tcatctgcac caccggtaaa
     3421 ctgccggttc cgtggccgac cctggttacc accctgacct acggtgttca gtgcttcgct
     3481 cgttacccgg accacatgaa acagcacgac ttcttcaaat ctgctatgcc ggaaggttac
     3541 gttcaggaac gtaccatctc tttcaaagac gacggtacct acaaaacccg tgctgaagtt
     3601 aaattcgaag gtgacaccct ggttaaccgt atcgaactga aaggtatcga cttcaaagaa
     3661 gacggtaaca tcctgggtca caaactggaa tacaacttca actctcacaa cgtttacatc
     3721 accgctgaca aacagaaaaa cggtatcaaa gctaacttca aaatccgtca caacgttgaa
     3781 gacggttctg ttcagctggc tgaccactac cagcagaaca ccccgatcgg tgacggtccg
     3841 gttctgctgc cggacaacca ctacctgtct acccagtctg ttctgtctaa agacccgaac
     3901 gaaaaacgtg accacatggt tctgctggaa ttcgttaccg ctgctggtat cacccacggt
     3961 atggacgaac tgtacaaaga tgcatgccag ttctagcata accctaatga gtgagctaac
     4021 ttacattaat tgcgttgcgc cttaattaac ggcactcctc agcaaatata atgaccctct
     4081 tgataaccca agagggcatt ttttaatgcc catggcgttt accacagcta acaccacgtc
     4141 gtccctatct gctgccctag gtctatgagt ggttgctgga taactttacg ggcatgcata
     4201 aggctcgtag gctatattca gggagaccac aacggtttcc ctctacaaat aattttgttt
     4261 aactttgaaa taaggaggta atacaaatgc cgcgtccgaa actgaaatct gacgacgaag
     4321 ttctggaagc ggcgaccgtt gttctgaaac gttgcggtcc gatcgaattc accctgtctg
     4381 gtgttgcgaa agaagttggt ctgtctcgtg cggcgctgat ccagcgtttc accaaccgtg
     4441 acaccctgct ggttcgtatg atggaacgtg gtgttgaaca ggttcgtcac tacctgaacg
     4501 cgatcccgat cggtgcgggt ccgcagggtc tgtgggaatt cctgcaggtt ctggttcgtt
     4561 ctatgaacac ccgtaacgac ttctctgtta actacctgat ctcttggtac gaactgcagg
     4621 ttccggaact gcgtaccctg gcgatccagc gtaaccgtgc ggttgttgaa ggtatccgta
     4681 aacgtctgcc gccgggtgcg ccggcggcgg cggaactgct gctgcactct gttatcgcgg
     4741 gtgcgaccat gcagtgggcg gttgacccgg acggtgaact ggcggaccac gttctggcgc
     4801 agatcgcggc gatcctgtgc ctgatgttcc cggaacacga cgacttccag ctgctgcagc
     4861 cgcacgcgta ataactagtg cttggattct caccaataaa aaacgcccgg cggcaaccga
     4921 gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc
     4981 caagcgagct cgtaaacttg gtctgacagc ccttaactta cttattaaat aatttatagc
     5041 tattgaaaag agataagaat tgttcaaagc taatattgtt taaatcgtca attcctgcat
     5101 gttttaagga attgttaaat tgattttttg taaatatttt cttgtattct ttgttaaccc
     5161 atttcataac gaaataatta tacttttgtt tatctttgtg tgatattctt gatttttttc
     5221 tacttaatct gataagtgag ctattcactt taggtttagg atgaaaatat tctcttggaa
     5281 ccatacttaa tatagaaata tcaacttctg ccattaaaag taatgccaat gagcgttttg
     5341 tatttaataa tcttttagca aacccgtatt ccacgattaa ataaatctca ttagctatac
     5401 tatcaaaaac aattttgcgt attatatccg tacttatgtt ataaggtata ttaccatata
     5461 ttttatagga ttggttttta ggaaatttaa actgcaatat atccttgttt aaaacttgga
     5521 aattatcgtg atcaacaagt ttattttctg tagttttgca taatttatgg tctatttcaa
     5581 tggcagttac gaaattacac ctctttacta attcaagggt aaaatggcct tttcctgagc
     5641 cgatttcaaa gatattatca tgttcattta atcttatatt tgtcattatt ttatctatat
     5701 tatgttttga agtaataaag ttttgactgt gttttatatt tttctcgttc atactcttcc
     5761 tttttcaat
//
