LOCUS       sequence-452545-        3460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sfGFP reporter for chimeric riboswitch with Cbe pfl ZTP riboswitch 
            aptamer and WT Pca rhtB ZTP riboswitch expression platform.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3460)
  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 3460)
  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: pJBL7345
FEATURES             Location/Qualifiers
     source          1..3460
                     /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(199..201,202..915)
                     /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             199..912
                     /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(229..257)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     852..872
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             935..964
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             980..997
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1053..1070)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1223..1269
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1422..2081
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1488..1507)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2102..2196
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2603..2620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2737..3282)
                     /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(2817..2836)
                     /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 ataacttact tttggagata agtcatgcct
      181 gatcacgtac atttccttat gagcaaagga gaagaacttt tcactggagt tgtcccaatt
      241 cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtccgtgg agagggtgaa
      301 ggtgatgcta caaacggaaa actcaccctt aaatttattt gcactactgg aaaactacct
      361 gttccgtggc caacacttgt cactactctg acctatggtg ttcaatgctt ttcccgttat
      421 ccggatcaca tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
      481 gaacgcacta tatctttcaa agatgacggg acctacaaga cgcgtgctga agtcaagttt
      541 gaaggtgata cccttgttaa tcgtatcgag ttaaagggta ttgattttaa agaagatgga
      601 aacattcttg gacacaaact cgagtacaac tttaactcac acaatgtata catcacggca
      661 gacaaacaaa agaatggaat caaagctaac ttcaaaattc gccacaacgt tgaagatggt
      721 tccgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
      781 ttaccagaca accattacct gtcgacacaa tctgtccttt cgaaagatcc caacgaaaag
      841 cgtgaccaca tggtccttct tgagtttgta actgctgctg ggattacaca tggcatggat
      901 gagctctaca aataaggatc tgaagcttgg gcccgaacaa aaactcatct cagaagagga
      961 tctgaatagc gccgtcgacc atcatcatca tcatcattga gtttaaacgg tctccagctt
     1021 ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga
     1081 agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc
     1141 atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg
     1201 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     1261 tcgttttatc tgttgtttgt cggtgaactg gatccttact cgagtctaga ctgcagttga
     1321 tcgggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat
     1381 tttttgagtt atcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
     1441 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     1501 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
     1561 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     1621 tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg atatgggata
     1681 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga
     1741 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt
     1801 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag
     1861 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct
     1921 tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc
     1981 tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg
     2041 aattacaaca gtactgcgat gagtggcagg gcggggcgta atttgatatc gagctcgctt
     2101 ggactcctgt tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg
     2161 ctcggttgcc gccgggcgtt ttttattggt gagaatccaa gcctccgatc aacgtctcat
     2221 tttcgccaaa agttggccca gggcttcccg gtatcaacag ggacaccagg atttatttat
     2281 tctgcgaagt gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct
     2341 gccaacttac tgatttagtg tatgatggtg tttttgaggt gctccagtgg cttctgtttc
     2401 tatcagctgt ccctcctgtt cagctactga cggggtggtg cgtaacggca aaagcaccgc
     2461 cggacatcag cgctagcgga gtgtatactg gcttactatg ttggcactga tgagggtgtc
     2521 agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt
     2581 gatacaggat atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc
     2641 ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac
     2701 ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga
     2761 caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag
     2821 ataccaggcg tttccccctg gcggctccct cgtgcgctct cctgttcctg cctttcggtt
     2881 taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg acactcagtt
     2941 ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt cagtccgacc
     3001 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac
     3061 cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc atgcgccggt
     3121 taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca gttacctcgg
     3181 ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg
     3241 ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca tcttattaat
     3301 cagataaaat atttctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
     3361 agccccatac gatataagtt gtaattctca tgtttgacag cttatcatcg ataagcttcc
     3421 gatggcgcgc cgagaggctt tacactttat gcttccggct
//
