LOCUS       sequence-102828-        5069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flavonoid biosensor. Low copy plasmid that expresses GFP 
            proportional to flavonoid concentration..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5069)
  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 5069)
  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-ttgR
FEATURES             Location/Qualifiers
     source          1..5069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(59..127,128..919)
                     /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(277..296)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(1072..2022)
                     /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      2070..2292
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     complement(2784..2801)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2954..3040
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3132..3159
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             3276..3989
                     /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             4282..4290
                     /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      4984..5031
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
       61 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      121 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      181 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      241 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      301 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      361 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      421 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
      481 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
      541 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
      601 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
      661 ccggcaacaa ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
      721 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtggctctcg
      781 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
      841 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
      901 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
      961 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1021 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgaac ctcagatcct
     1081 tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa
     1141 cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct caaaactggt
     1201 gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tatgtaggta
     1261 ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc
     1321 aagttcggtt actagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt
     1381 cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact
     1441 tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat
     1501 taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg
     1561 tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt
     1621 aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc
     1681 ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa
     1741 aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct
     1801 ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca tctgagcgta
     1861 ttggttataa gtgaacgata ccgtccgttc tttccttgta gggttttcaa tcgtggggtt
     1921 gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact
     1981 aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag
     2041 gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac tagtcctttt
     2101 cctttgagtt gtgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc
     2161 tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt
     2221 ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc
     2281 tgtgtataac tcactacttt agtcagttcc gcagtattac aaaaggatgt cgcaaacgct
     2341 gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag
     2401 ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc
     2461 tttttcgtga cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc
     2521 actacaggcg ccttttatgg attcatgcaa ggaaactacc cataatacaa gaaaagcccg
     2581 tcacgggctt ctcagggcgt tttatggcgg gtctgctatg tggtgctatc tgactttttg
     2641 ctgttcagca gttcctgccc tctgattttc cagtctgacc acttcggatt atcccgtgac
     2701 aggtcattca gactggctaa tgcacccagt aaggcagcgg tatcatcaac tggctgtttt
     2761 ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg
     2821 ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac
     2881 tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg
     2941 aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     3001 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     3061 cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca
     3121 tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctgct
     3181 agcaagtaag gccgaccacc cagcagtatt tacaaacaac catgaatgta agtatattcc
     3241 ttagcaacat ttaactttaa gaaggagata tacatatgcg taaaggtgaa gaactgttca
     3301 ccggtgttgt tccgatcctg gttgaactgg acggtgacgt taacggtcac aaattctctg
     3361 ttcgtggtga aggtgaaggt gacgctacca acggtaaact gaccctgaaa ttcatctgca
     3421 ccaccggtaa actgccggtt ccgtggccga ccctggttac caccctgacc tacggtgttc
     3481 agtgcttcgc tcgttacccg gaccacatga aacagcacga cttcttcaaa tctgctatgc
     3541 cggaaggtta cgttcaggaa cgtaccatct ctttcaaaga cgacggtacc tacaaaaccc
     3601 gtgctgaagt taaattcgaa ggtgacaccc tggttaaccg tatcgaactg aaaggtatcg
     3661 acttcaaaga agacggtaac atcctgggtc acaaactgga atacaacttc aactctcaca
     3721 acgtttacat caccgctgac aaacagaaaa acggtatcaa agctaacttc aaaatccgtc
     3781 acaacgttga agacggttct gttcagctgg ctgaccacta ccagcagaac accccgatcg
     3841 gtgacggtcc ggttctgctg ccggacaacc actacctgtc tacccagtct gttctgtcta
     3901 aagacccgaa cgaaaaacgt gaccacatgg ttctgctgga attcgttacc gctgctggta
     3961 tcacccacgg tatggacgaa ctgtacaaag atgcatgcca gttctagcat aaccctaatg
     4021 agtgagctaa cttacattaa ttgcgttgcg ccttaattaa cggcactcct cagcaaatat
     4081 aatgaccctc ttgataaccc aagagggcat tttttaatgc ccatggcgtt taccacagct
     4141 aacaccacgt cgtccctatc tgctgcccta ggtctatgag tggttgctgg ataactttac
     4201 gggcatgcat aaggctcgta ggctatattc agggagacca caacggtttc cctctacaaa
     4261 taattttgtt taactttgaa ataaggaggt aatacaaatg gtgcgtcgca ccaaagaaga
     4321 agcacaggaa acgcgtgcgc agattatcga agcggccgaa cgcgcgtttt ataaacgtgg
     4381 tgtggcacgt accacgctgg cagatattgc agaactggca ggtgttaccc gcggtgcaat
     4441 ctactggcat ttcaacaata aagccgaact ggttcaggca ctgctggatt ctctgcacga
     4501 aacgcatgat cacctggccc gtgcaagcga atctgaagat gaactggacc cgctgggctg
     4561 catgcgcaaa ctgctgctgc aggtgtttaa cgaactggtt ctggatgcac gtacccgtcg
     4621 cattaatgaa atcctgcatc acaaatgcga atttacggat gatatgtgtg aaattcgtca
     4681 gcagcgccag agcgccgtgc tggattgtca taaaggtatc accctggcac tggcaaacgc
     4741 agttcgtcgc ggtcagctgc cgggtgaact ggatgtggaa cgcgcagcgg ttgcgatgtt
     4801 tgcctatgtg gatggcctga ttggtcgttg gctgctgctg ccggatagtg ttgacctgct
     4861 gggcgatgtg gaaaaatggg ttgataccgg tctggatatg ctgcgtctga gcccggcgct
     4921 gcgcaaataa taagcgacta aaaaattgaa tgtaggaaac caacatgcca gttcgagcaa
     4981 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaacct
     5041 caggcatttg agaagcacac ggtcacact
//
