LOCUS       sequence-379068-        3465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Clostridium beijerinckii pfl ZTP riboswitch with sfGFP reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3465)
  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 3465)
  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: pJBL3907
FEATURES             Location/Qualifiers
     source          1..3465
                     /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(257..259,260..973)
                     /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             257..970
                     /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(287..315)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     910..930
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             993..1022
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1038..1055
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1111..1128)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1281..1327
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1480..2139
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1546..1565)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2160..2254
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2661..2678)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2795..3340)
                     /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(2875..2894)
                     /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 cgtcggcttt tttatatgga
      241 aaaggaggaa ggatctatga gcaaaggaga agaacttttc actggagttg tcccaattct
      301 tgttgaatta gatggtgatg ttaatgggca caaattttct gtccgtggag agggtgaagg
      361 tgatgctaca aacggaaaac tcacccttaa atttatttgc actactggaa aactacctgt
      421 tccgtggcca acacttgtca ctactctgac ctatggtgtt caatgctttt cccgttatcc
      481 ggatcacatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga
      541 acgcactata tctttcaaag atgacgggac ctacaagacg cgtgctgaag tcaagtttga
      601 aggtgatacc cttgttaatc gtatcgagtt aaagggtatt gattttaaag aagatggaaa
      661 cattcttgga cacaaactcg agtacaactt taactcacac aatgtataca tcacggcaga
      721 caaacaaaag aatggaatca aagctaactt caaaattcgc cacaacgttg aagatggttc
      781 cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt
      841 accagacaac cattacctgt cgacacaatc tgtcctttcg aaagatccca acgaaaagcg
      901 tgaccacatg gtccttcttg agtttgtaac tgctgctggg attacacatg gcatggatga
      961 gctctacaaa taaggatctg aagcttgggc ccgaacaaaa actcatctca gaagaggatc
     1021 tgaatagcgc cgtcgaccat catcatcatc atcattgagt ttaaacggtc tccagcttgg
     1081 ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag
     1141 cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat
     1201 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag
     1261 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     1321 gttttatctg ttgtttgtcg gtgaactgga tccttactcg agtctagact gcagttgatc
     1381 gggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt
     1441 tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
     1501 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
     1561 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     1621 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     1681 aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt
     1741 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     1801 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     1861 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     1921 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     1981 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     2041 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     2101 ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttgatatcga gctcgcttgg
     2161 actcctgttg atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct
     2221 cggttgccgc cgggcgtttt ttattggtga gaatccaagc ctccgatcaa cgtctcattt
     2281 tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat ttatttattc
     2341 tgcgaagtga tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc
     2401 caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct tctgtttcta
     2461 tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg
     2521 gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag
     2581 tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga
     2641 tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
     2701 cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt
     2761 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca
     2821 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat
     2881 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta
     2941 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc
     3001 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc
     3061 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca
     3121 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta
     3181 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt
     3241 caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt
     3301 ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca
     3361 gataaaatat ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag
     3421 ccccatacga tataagttgt aattctcatg tttgacagct tatca
//
