LOCUS       sequence-379071-        3473 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Synthetic OFF-logic Clostridium beijerinckii pfl ZTP riboswitch with
            sfGFP reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3473)
  AUTHORS   Bushhouse DZ, Lucks JB
  TITLE     Tuning strand displacement kinetics enables programmable ZTP 
            riboswitch dynamic range in vivo.
  JOURNAL   Nucleic Acids Res. 2023 Mar 2:gkad110. doi: 10.1093/nar/gkad110.
  PUBMED    36864761
REFERENCE   2  (bases 1 to 3473)
  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. 2023 Mar 2:gkad110. doi: 10.1093/nar/gkad110."
COMMENT     Sequence Label: pJBL6392
FEATURES             Location/Qualifiers
     source          1..3473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        69..103
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     CDS             join(265..267,268..981)
                     /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             265..978
                     /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(295..323)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     918..938
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1001..1030
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1046..1063
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1119..1136)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1289..1335
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1488..2147
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1554..1573)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2168..2262
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2669..2686)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2803..3348)
                     /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(2883..2902)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcgataagct tccgatggcg cgccgagagg ctttacactt tatgcttccg gctgaattct
       61 aaagatcttt gacagctagc tcagtcctag gtataatact agtatattag atattagtca
      121 tatgactgac ggaagtggag ttaccacatg aagtatgact aggcatatta tcttatatgc
      181 cacaaaaagc cgaccgtctg ggcaaaaaaa gcctggattg cgtcggcggt ccaggttttt
      241 tatatggaaa aggaggaagg atctatgagc aaaggagaag aacttttcac tggagttgtc
      301 ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt ccgtggagag
      361 ggtgaaggtg atgctacaaa cggaaaactc acccttaaat ttatttgcac tactggaaaa
      421 ctacctgttc cgtggccaac acttgtcact actctgacct atggtgttca atgcttttcc
      481 cgttatccgg atcacatgaa acggcatgac tttttcaaga gtgccatgcc cgaaggttat
      541 gtacaggaac gcactatatc tttcaaagat gacgggacct acaagacgcg tgctgaagtc
      601 aagtttgaag gtgataccct tgttaatcgt atcgagttaa agggtattga ttttaaagaa
      661 gatggaaaca ttcttggaca caaactcgag tacaacttta actcacacaa tgtatacatc
      721 acggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattcgcca caacgttgaa
      781 gatggttccg ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct
      841 gtccttttac cagacaacca ttacctgtcg acacaatctg tcctttcgaa agatcccaac
      901 gaaaagcgtg accacatggt ccttcttgag tttgtaactg ctgctgggat tacacatggc
      961 atggatgagc tctacaaata aggatctgaa gcttgggccc gaacaaaaac tcatctcaga
     1021 agaggatctg aatagcgccg tcgaccatca tcatcatcat cattgagttt aaacggtctc
     1081 cagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa
     1141 cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct
     1201 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc
     1261 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
     1321 ggcctttcgt tttatctgtt gtttgtcggt gaactggatc cttactcgag tctagactgc
     1381 agttgatcgg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg
     1441 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
     1501 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     1561 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     1621 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     1681 gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag ctggtgatat
     1741 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     1801 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     1861 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     1921 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     1981 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     2041 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
     2101 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt gatatcgagc
     2161 tcgcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
     2221 agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcct ccgatcaacg
     2281 tctcattttc gccaaaagtt ggcccagggc ttcccggtat caacagggac accaggattt
     2341 atttattctg cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt
     2401 gatgctgcca acttactgat ttagtgtatg atggtgtttt tgaggtgctc cagtggcttc
     2461 tgtttctatc agctgtccct cctgttcagc tactgacggg gtggtgcgta acggcaaaag
     2521 caccgccgga catcagcgct agcggagtgt atactggctt actatgttgg cactgatgag
     2581 ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga
     2641 atatgtgata caggatatat tccgcttcct cgctcactga ctcgctacgc tcggtcgttc
     2701 gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa gatgccagga
     2761 agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata ggctccgccc
     2821 ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc cgacaggact
     2881 ataaagatac caggcgtttc cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt
     2941 tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca cgcctgacac
     3001 tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc cccgttcagt
     3061 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa agacatgcaa
     3121 aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt gaagtcatgc
     3181 gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc aagccagtta
     3241 cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg caaggcggtt
     3301 ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga agatcatctt
     3361 attaatcaga taaaatattt ctagatttca gtgcaattta tctcttcaaa tgtagcacct
     3421 gaagtcagcc ccatacgata taagttgtaa ttctcatgtt tgacagctta tca
//
