LOCUS       sequence-266729-        3204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluoride riboswitch-regulated sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3204)
  AUTHORS   Thavarajah W, Silverman AD, Verosloff M, Kelley-Loughnane N, Jewett 
            MC, Lucks JB
  TITLE     Point-of-Use Detection of Environmental Fluoride via a Cell-Free 
            Riboswitch-Based Biosensor.
  JOURNAL   ACS Synth Biol. 2019 Dec 12. doi: 10.1021/acssynbio.9b00347.
  PUBMED    31829623
REFERENCE   2  (bases 1 to 3204)
  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. 2019 Dec 12. doi: 10.1021/acssynbio.9b00347."
COMMENT     Sequence Label: pJBL3752
FEATURES             Location/Qualifiers
     source          1..3204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(68..613)
                     /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(148..167)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        807..841
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     CDS             join(938..940,941..1654)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             938..1651
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(968..996)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1591..1611
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1687..1758
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1774..1801
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1957..2616
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2023..2042)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2637..2731
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3138..3155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
       61 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      121 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
      181 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
      241 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
      301 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
      361 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
      421 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
      481 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
      541 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
      601 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca
      661 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
      721 atgtttgaca gcttatcatc gataagcttc cgatggcgcg ccgagaggct ttacacttta
      781 tgcttccggc tgaattctaa agatctttga cagctagctc agtcctaggt ataatactag
      841 tttataggcg atggagttcg ccataaacgc tgcttagcta atgactccta ccagtatcac
      901 tactggtagg agtctatttt tttaggagga aggatctatg agcaaaggag aagaactttt
      961 cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc
     1021 tgtccgtgga gagggtgaag gtgatgctac aaacggaaaa ctcaccctta aatttatttg
     1081 cactactgga aaactacctg ttccgtggcc aacacttgtc actactctga cctatggtgt
     1141 tcaatgcttt tcccgttatc cggatcacat gaaacggcat gactttttca agagtgccat
     1201 gcccgaaggt tatgtacagg aacgcactat atctttcaaa gatgacggga cctacaagac
     1261 gcgtgctgaa gtcaagtttg aaggtgatac ccttgttaat cgtatcgagt taaagggtat
     1321 tgattttaaa gaagatggaa acattcttgg acacaaactc gagtacaact ttaactcaca
     1381 caatgtatac atcacggcag acaaacaaaa gaatggaatc aaagctaact tcaaaattcg
     1441 ccacaacgtt gaagatggtt ccgttcaact agcagaccat tatcaacaaa atactccaat
     1501 tggcgatggc cctgtccttt taccagacaa ccattacctg tcgacacaat ctgtcctttc
     1561 gaaagatccc aacgaaaagc gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg
     1621 gattacacat ggcatggatg agctctacaa ataaggatcc aaactcgagt aaggatctcc
     1681 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     1741 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
     1801 gtttatacct agggtacggt tttgggatcc ttactcgagt ctagactgca gttgatcggg
     1861 cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
     1921 gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
     1981 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
     2041 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
     2101 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
     2161 gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
     2221 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa
     2281 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
     2341 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat
     2401 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
     2461 cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
     2521 attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta
     2581 caacagtact gcgatgagtg gcagggcggg gcgtaatttg atatcgagct cgcttggact
     2641 cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg
     2701 ttgccgccgg gcgtttttta ttggtgagaa tccaagcctc cgatcaacgt ctcattttcg
     2761 ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc
     2821 gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa
     2881 cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct gtttctatca
     2941 gctgtccctc ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc accgccggac
     3001 atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga
     3061 agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac
     3121 aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     3181 gcggaaatgg cttacgaacg gggc
//
