LOCUS       sequence-403407-        3204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacillus cereus (B. ce) fluoride riboswitch, pseudoknot/terminator 
            overlap from T. pe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3204)
  AUTHORS   Hertz LM, White EN, Kuznedelov K, Cheng L, Yu AM, Kakkaramadam R, 
            Severinov K, Chen A, Lucks JB
  TITLE     The effect of pseudoknot base pairing on cotranscriptional 
            structural switching of the fluoride riboswitch.
  JOURNAL   Nucleic Acids Res. 2024 May 8;52(8):4466-4482. doi: 
            10.1093/nar/gkae231.
  PUBMED    38567721
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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2024-05-8- 8"; volume: "52"; issue: "8";
            pages: "4466-4482"
COMMENT     Sequence Label: pJBL814
FEATURES             Location/Qualifiers
     source          1..3204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     175..192
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(599..693)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(714..1373)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1288..1307
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(1529..1556)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(1572..1643)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(1676..2389,2390..2392))
                     /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             complement(1679..2392)
                     /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(1719..1739)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     2334..2362
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     promoter        complement(2489..2523)
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     rep_origin      join(2717..3204,1..58)
                     /direction=RIGHT
                     /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     3163..3182
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa
       61 cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc ttccaggaaa
      121 tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga gcgtagcgag
      181 tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg caccggtgca
      241 gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg ccaacatagt
      301 aagccagtat acactccgct agcgctgatg tccggcggtg cttttgccgt tacgcaccac
      361 cccgtcagta gctgaacagg agggacagct gatagaaaca gaagccactg gagcacctca
      421 aaaacaccat catacactaa atcagtaagt tggcagcatc acccgacgca ctttgcgccg
      481 aataaatacc tgtgacggaa gatcacttcg cagaataaat aaatcctggt gtccctgttg
      541 ataccgggaa gccctgggcc aacttttggc gaaaatgaga cgttgatcgg aggcttggat
      601 tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt
      661 ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgatatc aaattacgcc
      721 ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag
      781 ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
      841 gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
      901 aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata
      961 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
     1021 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt
     1081 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
     1141 ttcattgcca tacgaaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
     1201 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
     1261 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
     1321 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cattttagct
     1381 tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca
     1441 ttatggtgaa agttggaacc tcttacgtgc ccgatcaact gcagtctaga ctcgagtaag
     1501 gatcccaaaa ccgtacccta ggtataaacg cagaaaggcc cacccgaagg tgagccagtg
     1561 tgactctagt agagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc
     1621 gactgagcct ttcgttttat ttgatgcctg gagatcctta ctcgagtttg gatccttatt
     1681 tgtagagctc atccatgcca tgtgtaatcc cagcagcagt tacaaactca agaaggacca
     1741 tgtggtcacg cttttcgttg ggatctttcg aaaggacaga ttgtgtcgac aggtaatggt
     1801 tgtctggtaa aaggacaggg ccatcgccaa ttggagtatt ttgttgataa tggtctgcta
     1861 gttgaacgga accatcttca acgttgtggc gaattttgaa gttagctttg attccattct
     1921 tttgtttgtc tgccgtgatg tatacattgt gtgagttaaa gttgtactcg agtttgtgtc
     1981 caagaatgtt tccatcttct ttaaaatcaa taccctttaa ctcgatacga ttaacaaggg
     2041 tatcaccttc aaacttgact tcagcacgcg tcttgtaggt cccgtcatct ttgaaagata
     2101 tagtgcgttc ctgtacataa ccttcgggca tggcactctt gaaaaagtca tgccgtttca
     2161 tgtgatccgg ataacgggaa aagcattgaa caccataggt cagagtagtg acaagtgttg
     2221 gccacggaac aggtagtttt ccagtagtgc aaataaattt aagggtgagt tttccgtttg
     2281 tagcatcacc ttcaccctct ccacggacag aaaatttgtg cccattaaca tcaccatcta
     2341 attcaacaag aattgggaca actccagtga aaagttcttc tcctttgctc atagatcctt
     2401 cctcctaaaa aaataggcct ctaccagtag tgatactggt agaggccatt agctaagcag
     2461 cgtttatggc gggcctcatc gcctataaac tagtattata cctaggactg agctagctgt
     2521 caaagatctt tagaattcag ccggaagcat aaagtgtaaa gcctctcggc gcgccatcgg
     2581 aagcttatcg atgataagct gtcaaacatg agaattacaa cttatatcgt atggggctga
     2641 cttcaggtgc tacatttgaa gagataaatt gcactgaaat ctagaaatat tttatctgat
     2701 taataagatg atcttcttga gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg
     2761 aaaaaaccgc cttgcagggc ggtttttcga aggttctctg agctaccaac tctttgaacc
     2821 gaggtaactg gcttggagga gcgcagtcac caaaacttgt cctttcagtt tagccttaac
     2881 cggcgcatga cttcaagact aactcctcta aatcaattac cagtggctgc tgccagtggt
     2941 gcttttgcat gtctttccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3001 tcggactgaa cggggggttc gtgcatacag tccagcttgg agcgaactgc ctacccggaa
     3061 ctgagtgtca ggcgtggaat gagacaaacg cggccataac agcggaatga caccggtaaa
     3121 ccgaaaggca ggaacaggag agcgcacgag ggagccgcca gggggaaacg cctggtatct
     3181 ttatagtcct gtcgggtttc gcca
//
