LOCUS       sequence-431670-        3255 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sfGFP reporter for synthetic transcriptional variant of Pca rhtB ZTP
            riboswitch.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3255)
  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 3255)
  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: pJBL7346
FEATURES             Location/Qualifiers
     source          1..3255
                     /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(198..200,201..914)
                     /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             198..911
                     /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(228..256)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     851..871
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      947..1018
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1034..1061
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1217..1876
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFKSVAKCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     terminator      1897..1991
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2398..2415)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2532..3077)
                     /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(2612..2631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaattctaaa gatctttgac agctagctca gtcctaggta taatactagt aatgcgctca
       61 tgttactgac gcttaaagat agatcactat cgtgaagcat gggtcattac gagccgcgcg
      121 tctgggcata acttagagtt gttcataagt catgcccaga cgcgcggctt ttttatatgg
      181 aaaaggagga aggatctatg agcaaaggag aagaactttt cactggagtt gtcccaattc
      241 ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtccgtgga gagggtgaag
      301 gtgatgctac aaacggaaaa ctcaccctta aatttatttg cactactgga aaactacctg
      361 ttccgtggcc aacacttgtc actactctga cctatggtgt tcaatgcttt tcccgttatc
      421 cggatcacat gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg
      481 aacgcactat atctttcaaa gatgacggga cctacaagac gcgtgctgaa gtcaagtttg
      541 aaggtgatac ccttgttaat cgtatcgagt taaagggtat tgattttaaa gaagatggaa
      601 acattcttgg acacaaactc gagtacaact ttaactcaca caatgtatac atcacggcag
      661 acaaacaaaa gaatggaatc aaagctaact tcaaaattcg ccacaacgtt gaagatggtt
      721 ccgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt
      781 taccagacaa ccattacctg tcgacacaat ctgtcctttc gaaagatccc aacgaaaagc
      841 gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg gattacacat ggcatggatg
      901 agctctacaa ataaggatcc aaactcgagt aaggatctcc aggcatcaaa taaaacgaaa
      961 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta
     1021 ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatacct agggtacggt
     1081 tttgggatcc ttactcgagt ctagactgca gttgatcggg cacgtaagag gttccaactt
     1141 tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg
     1201 agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca
     1261 atggcatcgt aaagaacatt ttgaggcatt taagtcagtt gctaaatgta cctataacca
     1321 gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt
     1381 ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg aatttcgtat
     1441 ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt
     1501 ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca
     1561 gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
     1621 taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag
     1681 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa
     1741 atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt
     1801 ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg
     1861 gcagggcggg gcgtaatttg atatcgagct cgcttggact cctgttgata gatccagtaa
     1921 tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta
     1981 ttggtgagaa tccaagcctc cgatcaacgt ctcattttcg ccaaaagttg gcccagggct
     2041 tcccggtatc aacagggaca ccaggattta tttattctgc gaagtgatct tccgtcacag
     2101 gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa cttactgatt tagtgtatga
     2161 tggtgttttt gaggtgctcc agtggcttct gtttctatca gctgtccctc atgttcagct
     2221 actgacgggg tggtgcgtaa cggcaaaagc accgccggac atcagcgcta gcggagtgta
     2281 tactggctta ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag
     2341 aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc
     2401 gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg
     2461 gggcggagat ttcctggaag atgccaggaa gataattaac agggaagtga gagggccgcg
     2521 gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca
     2581 aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
     2641 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
     2701 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
     2761 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
     2821 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
     2881 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
     2941 ttggtgactg tgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
     3001 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
     3061 gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc tagatttcag
     3121 tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat
     3181 tatcatgttt gacagcttat catcgataag cttccgatgg cgcgccgaga ggctttacac
     3241 tttatgcttc cggct
//
