LOCUS       sequence-437374-        3208 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sfGFP reporter for Bce crcB fluoride riboswitch.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3208)
  AUTHORS   Bushhouse DZ, Fu J, Lucks JB
  TITLE     RNA folding kinetics control riboswitch sensitivity in vivo
  JOURNAL   bioRxiv 2024.03.29.587317
REFERENCE   2  (bases 1 to 3208)
  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: "bioRxiv 
            2024.03.29.587317"
COMMENT     Sequence Label: pJBL7368
FEATURES             Location/Qualifiers
     source          1..3208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..50
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     CDS             join(151..153,154..867)
                     /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             151..864
                     /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(181..209)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     804..824
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      900..971
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      987..1014
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1170..1829
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1236..1255)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1850..1944
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2351..2368)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2485..3030)
                     /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(2565..2584)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaattctaaa gatctttgac agctagctca gtcctaggta taatactagt ataattatag
       61 gcgatggagt tcgccataaa cgctgcttag ctaatgactc ctaccagtat cactactggt
      121 aggagtctat ttttttagga ggaaggatct atgagcaaag gagaagaact tttcactgga
      181 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtccgt
      241 ggagagggtg aaggtgatgc tacaaacgga aaactcaccc ttaaatttat ttgcactact
      301 ggaaaactac ctgttccgtg gccaacactt gtcactactc tgacctatgg tgttcaatgc
      361 ttttcccgtt atccggatca catgaaacgg catgactttt tcaagagtgc catgcccgaa
      421 ggttatgtac aggaacgcac tatatctttc aaagatgacg ggacctacaa gacgcgtgct
      481 gaagtcaagt ttgaaggtga tacccttgtt aatcgtatcg agttaaaggg tattgatttt
      541 aaagaagatg gaaacattct tggacacaaa ctcgagtaca actttaactc acacaatgta
      601 tacatcacgg cagacaaaca aaagaatgga atcaaagcta acttcaaaat tcgccacaac
      661 gttgaagatg gttccgttca actagcagac cattatcaac aaaatactcc aattggcgat
      721 ggccctgtcc ttttaccaga caaccattac ctgtcgacac aatctgtcct ttcgaaagat
      781 cccaacgaaa agcgtgacca catggtcctt cttgagtttg taactgctgc tgggattaca
      841 catggcatgg atgagctcta caaataagga tccaaactcg agtaaggatc tccaggcatc
      901 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
      961 tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata
     1021 cctagggtac ggttttggga tccttactcg agtctagact gcagttgatc gggcacgtaa
     1081 gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat
     1141 cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg
     1201 ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat
     1261 gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa
     1321 ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc
     1381 cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt
     1441 gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg
     1501 acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc
     1561 tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg
     1621 tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt
     1681 tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg
     1741 ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt
     1801 actgcgatga gtggcagggc ggggcgtaat ttgatatcga gctcgcttgg actcctgttg
     1861 atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc
     1921 cgggcgtttt ttattggtga gaatccaagc ctccgatcaa cgtctcattt tcgccaaaag
     1981 ttggcccagg gcttcccggt atcaacaggg acaccaggat ttatttattc tgcgaagtga
     2041 tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc caacttactg
     2101 atttagtgta tgatggtgtt tttgaggtgc tccagtggct tctgtttcta tcagctgtcc
     2161 ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg gacatcagcg
     2221 ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag tgaagtgctt
     2281 catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga tacaggatat
     2341 attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa
     2401 tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt aacagggaag
     2461 tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca agcatcacga
     2521 aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2581 tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat
     2641 tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag
     2701 ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat
     2761 ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag
     2821 ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact
     2881 gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg
     2941 gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa
     3001 gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca gataaaatat
     3061 ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga
     3121 tataagttgt aattctcatg tttgacagct tatcatcgat aagcttccga tggcgcgccg
     3181 agaggcttta cactttatgc ttccggct
//
