LOCUS       sequence-379064-        3464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Clostridium beijerinckii pfl ZTP riboswitch A26∆ OFF control with 
            sfGFP reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3464)
  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 3464)
  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: pJBL6090
FEATURES             Location/Qualifiers
     source          1..3464
                     /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(256..258,259..972)
                     /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             256..969
                     /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(286..314)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     909..929
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             992..1021
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1037..1054
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1110..1127)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1280..1326
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1479..2138
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1545..1564)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2159..2253
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2660..2677)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2794..3339)
                     /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(2874..2893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcgataagct tccgatggcg cgccgagagg ctttacactt tatgcttccg gctgaattct
       61 aaagatcttt gacagctagc tcagtcctag gtataatact agtatattag atattagtca
      121 tatgactgcg gaagtggagt taccacatga agtatgacta ggcatattat cttatatgcc
      181 acaaaaagcc gaccgtctgg gcaaaaaaag cctggattgc gtcggctttt ttatatggaa
      241 aaggaggaag gatctatgag caaaggagaa gaacttttca ctggagttgt cccaattctt
      301 gttgaattag atggtgatgt taatgggcac aaattttctg tccgtggaga gggtgaaggt
      361 gatgctacaa acggaaaact cacccttaaa tttatttgca ctactggaaa actacctgtt
      421 ccgtggccaa cacttgtcac tactctgacc tatggtgttc aatgcttttc ccgttatccg
      481 gatcacatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa
      541 cgcactatat ctttcaaaga tgacgggacc tacaagacgc gtgctgaagt caagtttgaa
      601 ggtgataccc ttgttaatcg tatcgagtta aagggtattg attttaaaga agatggaaac
      661 attcttggac acaaactcga gtacaacttt aactcacaca atgtatacat cacggcagac
      721 aaacaaaaga atggaatcaa agctaacttc aaaattcgcc acaacgttga agatggttcc
      781 gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta
      841 ccagacaacc attacctgtc gacacaatct gtcctttcga aagatcccaa cgaaaagcgt
      901 gaccacatgg tccttcttga gtttgtaact gctgctggga ttacacatgg catggatgag
      961 ctctacaaat aaggatctga agcttgggcc cgaacaaaaa ctcatctcag aagaggatct
     1021 gaatagcgcc gtcgaccatc atcatcatca tcattgagtt taaacggtct ccagcttggc
     1081 tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc
     1141 ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     1201 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga
     1261 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1321 ttttatctgt tgtttgtcgg tgaactggat ccttactcga gtctagactg cagttgatcg
     1381 ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt
     1441 ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     1501 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     1561 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     1621 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     1681 atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg
     1741 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     1801 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     1861 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
     1921 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     1981 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     2041 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
     2101 tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tgatatcgag ctcgcttgga
     2161 ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc
     2221 ggttgccgcc gggcgttttt tattggtgag aatccaagcc tccgatcaac gtctcatttt
     2281 cgccaaaagt tggcccaggg cttcccggta tcaacaggga caccaggatt tatttattct
     2341 gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc
     2401 aacttactga tttagtgtat gatggtgttt ttgaggtgct ccagtggctt ctgtttctat
     2461 cagctgtccc tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg
     2521 acatcagcgc tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt
     2581 gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat
     2641 acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc
     2701 gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta
     2761 acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa
     2821 gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata
     2881 ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac
     2941 cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg
     3001 ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct
     3061 gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac
     3121 tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa
     3181 ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc
     3241 aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt
     3301 tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaatcag
     3361 ataaaatatt tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc
     3421 cccatacgat ataagttgta attctcatgt ttgacagctt atca
//
