LOCUS       sequence-433102-        3484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sfGFP reporter for chimeric riboswitch with Cbe pfl ZTP riboswitch 
            aptamer and transcriptional variant of Pca rhtB ZTP riboswitch 
            expression platform.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3484)
  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 3484)
  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: pJBL7316
FEATURES             Location/Qualifiers
     source          1..3484
                     /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(223..225,226..939)
                     /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             223..936
                     /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(253..281)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     876..896
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             959..988
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1004..1021
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1077..1094)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1247..1293
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1446..2105
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1512..1531)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2126..2220
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2627..2644)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2761..3306)
                     /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(2841..2860)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaattctaaa gatctttgac agctagctca gtcctaggta taatactagt atattagata
       61 ttagtcatat gactgacgga agtggagtta ccacatgaag tatgactagg catattatct
      121 tatatgccac aaaaagccga ccgtctgggc ataacttaga gttgttcata agtcatgcct
      181 ggattgcgtc ggctttttta tatggaaaag gaggaaggat ctatgagcaa aggagaagaa
      241 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa
      301 ttttctgtcc gtggagaggg tgaaggtgat gctacaaacg gaaaactcac ccttaaattt
      361 atttgcacta ctggaaaact acctgttccg tggccaacac ttgtcactac tctgacctat
      421 ggtgttcaat gcttttcccg ttatccggat cacatgaaac ggcatgactt tttcaagagt
      481 gccatgcccg aaggttatgt acaggaacgc actatatctt tcaaagatga cgggacctac
      541 aagacgcgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatcgtat cgagttaaag
      601 ggtattgatt ttaaagaaga tggaaacatt cttggacaca aactcgagta caactttaac
      661 tcacacaatg tatacatcac ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa
      721 attcgccaca acgttgaaga tggttccgtt caactagcag accattatca acaaaatact
      781 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtcgac acaatctgtc
      841 ctttcgaaag atcccaacga aaagcgtgac cacatggtcc ttcttgagtt tgtaactgct
      901 gctgggatta cacatggcat ggatgagctc tacaaataag gatctgaagc ttgggcccga
      961 acaaaaactc atctcagaag aggatctgaa tagcgccgtc gaccatcatc atcatcatca
     1021 ttgagtttaa acggtctcca gcttggctgt tttggcggat gagagaagat tttcagcctg
     1081 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
     1141 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
     1201 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     1261 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga actggatcct
     1321 tactcgagtc tagactgcag ttgatcgggc acgtaagagg ttccaacttt caccataatg
     1381 aaataagatc actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag
     1441 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     1501 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     1561 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     1621 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag
     1681 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     1741 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     1801 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     1861 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     1921 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     1981 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     2041 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     2101 cgtaatttga tatcgagctc gcttggactc ctgttgatag atccagtaat gacctcagaa
     2161 ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat
     2221 ccaagcctcc gatcaacgtc tcattttcgc caaaagttgg cccagggctt cccggtatca
     2281 acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg
     2341 gcgcaaagtg cgtcgggtga tgctgccaac ttactgattt agtgtatgat ggtgtttttg
     2401 aggtgctcca gtggcttctg tttctatcag ctgtccctcc tgttcagcta ctgacggggt
     2461 ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac
     2521 tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg
     2581 caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact
     2641 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
     2701 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
     2761 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
     2821 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
     2881 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
     2941 tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc
     3001 acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3061 acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag
     3121 ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc
     3181 gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa
     3241 ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga
     3301 tctcaagaag atcatcttat taatcagata aaatatttct agatttcagt gcaatttatc
     3361 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgtttg
     3421 acagcttatc atcgataagc ttccgatggc gcgccgagag gctttacact ttatgcttcc
     3481 ggct
//
