LOCUS       sequence-437428-        3554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sfGFP reporter for Bsu yxjA 2AP riboswitch.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3554)
  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 3554)
  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: pJBL730
FEATURES             Location/Qualifiers
     source          1..3554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        37..71
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     CDS             join(314..316,317..1030)
                     /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             314..1027
                     /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(344..372)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     967..987
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1050..1079
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1095..1112
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1168..1185)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1338..1384
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             1537..2196
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1603..1622)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2217..2311
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2718..2735)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2852..3397)
                     /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(2932..2951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttacacttta tgcttccggc ttgattctaa agatctttga cagctagctc agtcctaggt
       61 ataatactag tatcttagaa aaagacattc ttgtatatga tcagtaatat ggtctgattg
      121 tttctaccta gtaaccgtaa aaaactagat tacaagaaag tttgaataaa tttgaacgag
      181 ttgaaaagga caagttcttt tctgtttgct cttatttttc acactttctg cacttccaga
      241 atttgtgaag gataagagct ttttttgttt ccataataac cctcatagga gttgctatca
      301 ggaggaagga tctatgagca aaggagaaga acttttcact ggagttgtcc caattcttgt
      361 tgaattagat ggtgatgtta atgggcacaa attttctgtc cgtggagagg gtgaaggtga
      421 tgctacaaac ggaaaactca cccttaaatt tatttgcact actggaaaac tacctgttcc
      481 gtggccaaca cttgtcacta ctctgaccta tggtgttcaa tgcttttccc gttatccgga
      541 tcacatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaacg
      601 cactatatct ttcaaagatg acgggaccta caagacgcgt gctgaagtca agtttgaagg
      661 tgataccctt gttaatcgta tcgagttaaa gggtattgat tttaaagaag atggaaacat
      721 tcttggacac aaactcgagt acaactttaa ctcacacaat gtatacatca cggcagacaa
      781 acaaaagaat ggaatcaaag ctaacttcaa aattcgccac aacgttgaag atggttccgt
      841 tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc
      901 agacaaccat tacctgtcga cacaatctgt cctttcgaaa gatcccaacg aaaagcgtga
      961 ccacatggtc cttcttgagt ttgtaactgc tgctgggatt acacatggca tggatgagct
     1021 ctacaaataa ggatctgaag cttgggcccg aacaaaaact catctcagaa gaggatctga
     1081 atagcgccgt cgaccatcat catcatcatc attgagttta aacggtctcc agcttggctg
     1141 ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg
     1201 tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc
     1261 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt
     1321 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     1381 ttatctgttg tttgtcggtg aactggatcc ttactcgagt ctagactgca gttgatcggg
     1441 cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
     1501 gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
     1561 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
     1621 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
     1681 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
     1741 gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
     1801 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa
     1861 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
     1921 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat
     1981 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
     2041 cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
     2101 attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta
     2161 caacagtact gcgatgagtg gcagggcggg gcgtaatttg atatcgagct cgcttggact
     2221 cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg
     2281 ttgccgccgg gcgtttttta ttggtgagaa tccaagcctc cgatcaacgt ctcattttcg
     2341 ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc
     2401 gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa
     2461 cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct gtttctatca
     2521 gctgtccctc ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc accgccggac
     2581 atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga
     2641 agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac
     2701 aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     2761 gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac
     2821 agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc
     2881 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc
     2941 aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg
     3001 gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg
     3061 taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc
     3121 gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg
     3181 gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg
     3241 ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa
     3301 agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc
     3361 agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat
     3421 aaaatatttc tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc
     3481 catacgatat aagttgtaat tctcatgttt gacagcttat catcgataag cttccgatgg
     3541 cgcgccgaga ggct
//
