LOCUS       sequence-431669-        3246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sfGFP reporter for Bsu pbuE* 2AP riboswitch.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3246)
  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 3246)
  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: pJBL6951
FEATURES             Location/Qualifiers
     source          1..3246
                     /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(189..191,192..905)
                     /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             189..902
                     /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(219..247)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     842..862
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      938..1009
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1025..1052
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1208..1867
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1274..1293)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1888..1982
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2389..2406)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2523..3068)
                     /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(2603..2622)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaattctaaa gatctttgac agctagctca gtcctaggta taatactagt aattaaatag
       61 acgtcattat cacttgtata acctcaataa tatggtttga gggtgtctac caggaaccgt
      121 aaaatcctga ttacaaaatt tgtttatgac attttttgta atcaggattt ttttaggagg
      181 aaggatctat gagcaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
      241 tagatggtga tgttaatggg cacaaatttt ctgtccgtgg agagggtgaa ggtgatgcta
      301 caaacggaaa actcaccctt aaatttattt gcactactgg aaaactacct gttccgtggc
      361 caacacttgt cactactctg acctatggtg ttcaatgctt ttcccgttat ccggatcaca
      421 tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaacgcacta
      481 tatctttcaa agatgacggg acctacaaga cgcgtgctga agtcaagttt gaaggtgata
      541 cccttgttaa tcgtatcgag ttaaagggta ttgattttaa agaagatgga aacattcttg
      601 gacacaaact cgagtacaac tttaactcac acaatgtata catcacggca gacaaacaaa
      661 agaatggaat caaagctaac ttcaaaattc gccacaacgt tgaagatggt tccgttcaac
      721 tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca
      781 accattacct gtcgacacaa tctgtccttt cgaaagatcc caacgaaaag cgtgaccaca
      841 tggtccttct tgagtttgta actgctgctg ggattacaca tggcatggat gagctctaca
      901 aataaggatc caaactcgag taaggatctc caggcatcaa ataaaacgaa aggctcagtc
      961 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
     1021 cactggctca ccttcgggtg ggcctttctg cgtttatacc tagggtacgg ttttgggatc
     1081 cttactcgag tctagactgc agttgatcgg gcacgtaaga ggttccaact ttcaccataa
     1141 tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttaag gagctaagga
     1201 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
     1261 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     1321 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     1381 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa
     1441 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
     1501 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
     1561 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     1621 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
     1681 aaacgtggcc aatatggaca acttcttcgc cccagttttc accatgggca aatattatac
     1741 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg
     1801 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     1861 ggcgtaattt gatatcgagc tcgcttggac tcctgttgat agatccagta atgacctcag
     1921 aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga
     1981 atccaagcct ccgatcaacg tctcattttc gccaaaagtt ggcccagggc ttcccggtat
     2041 caacagggac accaggattt atttattctg cgaagtgatc ttccgtcaca ggtatttatt
     2101 cggcgcaaag tgcgtcgggt gatgctgcca acttactgat ttagtgtatg atggtgtttt
     2161 tgaggtgctc cagtggcttc tgtttctatc agctgtccct cctgttcagc tactgacggg
     2221 gtggtgcgta acggcaaaag caccgccgga catcagcgct agcggagtgt atactggctt
     2281 actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc
     2341 tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga
     2401 ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga
     2461 tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg
     2521 tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg
     2581 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg
     2641 cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg
     2701 tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat
     2761 gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2821 caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg
     2881 agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact
     2941 gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa
     3001 aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac
     3061 gatctcaaga agatcatctt attaatcaga taaaatattt ctagatttca gtgcaattta
     3121 tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa ttctcatgtt
     3181 tgacagctta tcatcgataa gcttccgatg gcgcgccgag aggctttaca ctttatgctt
     3241 ccggct
//
