LOCUS       sequence-431668-        3233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sfGFP reporter for WT Pca rhtB ZTP riboswitch.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3233)
  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 3233)
  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: pJBL6931
FEATURES             Location/Qualifiers
     source          1..3233
                     /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(176..178,179..892)
                     /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             176..889
                     /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(206..234)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     829..849
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      925..996
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1012..1039
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1195..1854
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFKSVAKCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     terminator      1875..1969
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2376..2393)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2510..3055)
                     /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(2590..2609)
                     /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 acttactttt ggagataagt catgcctgat cacgtacatt tccttatgag
      181 caaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt
      241 taatgggcac aaattttctg tccgtggaga gggtgaaggt gatgctacaa acggaaaact
      301 cacccttaaa tttatttgca ctactggaaa actacctgtt ccgtggccaa cacttgtcac
      361 tactctgacc tatggtgttc aatgcttttc ccgttatccg gatcacatga aacggcatga
      421 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga
      481 tgacgggacc tacaagacgc gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg
      541 tatcgagtta aagggtattg attttaaaga agatggaaac attcttggac acaaactcga
      601 gtacaacttt aactcacaca atgtatacat cacggcagac aaacaaaaga atggaatcaa
      661 agctaacttc aaaattcgcc acaacgttga agatggttcc gttcaactag cagaccatta
      721 tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc
      781 gacacaatct gtcctttcga aagatcccaa cgaaaagcgt gaccacatgg tccttcttga
      841 gtttgtaact gctgctggga ttacacatgg catggatgag ctctacaaat aaggatccaa
      901 actcgagtaa ggatctccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
      961 tttcgtttta tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct
     1021 tcgggtgggc ctttctgcgt ttatacctag ggtacggttt tgggatcctt actcgagtct
     1081 agactgcagt tgatcgggca cgtaagaggt tccaactttc accataatga aataagatca
     1141 ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatggag
     1201 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     1261 gaggcattta agtcagttgc taaatgtacc tataaccaga ccgttcagct ggatattacg
     1321 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     1381 cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga cggtgagctg
     1441 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     1501 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
     1561 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     1621 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     1681 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
     1741 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
     1801 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaatttgat
     1861 atcgagctcg cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga
     1921 tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc caagcctccg
     1981 atcaacgtct cattttcgcc aaaagttggc ccagggcttc ccggtatcaa cagggacacc
     2041 aggatttatt tattctgcga agtgatcttc cgtcacaggt atttattcgg cgcaaagtgc
     2101 gtcgggtgat gctgccaact tactgattta gtgtatgatg gtgtttttga ggtgctccag
     2161 tggcttctgt ttctatcagc tgtccctcat gttcagctac tgacggggtg gtgcgtaacg
     2221 gcaaaagcac cgccggacat cagcgctagc ggagtgtata ctggcttact atgttggcac
     2281 tgatgagggt gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt
     2341 cagcagaata tgtgatacag gatatattcc gcttcctcgc tcactgactc gctacgctcg
     2401 gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat
     2461 gccaggaaga taattaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc
     2521 tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga
     2581 caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc
     2641 ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc
     2701 ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc
     2761 gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga
     2821 catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa
     2881 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag
     2941 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa
     3001 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga
     3061 tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct cttcaaatgt
     3121 agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgtttga cagcttatca
     3181 tcgataagct tccgatggcg cgccgagagg ctttacactt tatgcttccg gct
//
