LOCUS       sequence-461089-        4626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gfpUV reporter vector with a high performance adenine-dependent 
            riboswitch.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4626)
  AUTHORS   Drogalis LK, Batey RT
  TITLE     Requirements for efficient ligand-gated co-transcriptional switching
            in designed variants of the B. subtilis pbuE adenine-responsive 
            riboswitch in E. coli.
  JOURNAL   PLoS One. 2020 Dec 1;15(12):e0243155. doi: 
            10.1371/journal.pone.0243155. eCollection 2020.
  PUBMED    33259551
REFERENCE   2  (bases 1 to 4626)
  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: "PLoS One.";
            date: "2020-12-1"; volume: "15(12):e0243155. doi"; pages: " 
            10.1371/journal.pone.0243155. eCollection 2020"
COMMENT     Sequence Label: pP4-A
FEATURES             Location/Qualifiers
     source          1..4626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      139..225
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      317..344
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             656..1372
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(686..714)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1309..1329
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1713..1731)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1799..1903
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1904..1972,1973..2764)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2122..2141)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2935..3523
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3424..3443
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3677..3694
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3709..3849
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3951..4142)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cccgagaatg cgcattgttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
       61 cgaactcaga agcgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
      121 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
      181 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
      241 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
      301 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgct agccacagct
      361 aacaccacgt cgtccctatc tgctgcccta ggtctatgag tggttgctgg ataactttac
      421 gggcatgcat aaggctcgta taatatattc cacttgtata acctcaataa tatggtttga
      481 gggtgtctac caggaaccgt aaaatcctga ttacaagccg ttttttcggc ttgtaatcag
      541 gatttttttt atttactagt acatttaagt aaaggagttt gttatgacca tgattacgcc
      601 aagcttgcat gcctgcaggt cgactctaga ggatccccgg gtaccggtag aaaaaatgag
      661 taaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt
      721 taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact
      781 tacccttaaa tttatttgca ctactggaaa actacctgtt ccatggccaa cacttgtcac
      841 tactttctct tatggtgttc aatgcttttc ccgttatccg gatcatatga aacggcatga
      901 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga
      961 tgacgggaac tacaagacgc gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg
     1021 tatcgagtta aaaggtattg attttaaaga agatggaaac attctcggac acaaactcga
     1081 gtacaactat aactcacaca atgtatacat cacggcagac aaacaaaaga atggaatcaa
     1141 agctaacttc aaaattcgcc acaacattga agatggatcc gttcaactag cagaccatta
     1201 tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc
     1261 gacacaatct gccctttcga aagatcccaa cgaaaagcgt gaccacatgg tccttcttga
     1321 gtttgtaact gctgctggga ttacacatgg catggatgag ctctacaaat aatgaattcc
     1381 aactgagcgc cggtcgctac cattaccaac ttgtctggtg tcaaaaataa taggcctact
     1441 agtcggccgt acgggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     1501 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     1561 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     1621 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     1681 aggagaaaat accgcatcag gcggccttaa gggcctcgtg atacgcctat ttttataggt
     1741 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     1801 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1861 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     1921 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     1981 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2041 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2101 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     2161 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2221 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2281 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2341 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2401 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     2461 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2521 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2581 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     2641 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2701 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2761 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2821 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2881 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2941 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3001 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3061 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     3121 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3181 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3241 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3301 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3361 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3421 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3481 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3541 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3601 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3661 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     3721 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
     3781 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     3841 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     3901 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     3961 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     4021 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     4081 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     4141 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     4201 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     4261 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     4321 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     4381 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     4441 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     4501 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     4561 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ccaggaccca
     4621 acgctg
//
