LOCUS       sequence-192124-        4617 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFPuv reporter that places expression of a fluorescent protein 
            reporter (GFPuv) under control of the env8HyCbl riboswitch. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4617)
  AUTHORS   Polaski JT, Holmstrom ED, Nesbitt DJ, Batey RT
  TITLE     Mechanistic Insights into Cofactor-Dependent Coupling of RNA Folding
            and mRNA Transcription/Translation by a Cobalamin Riboswitch.
  JOURNAL   Cell Rep. 2016 May 3;15(5):1100-10. doi: 
            10.1016/j.celrep.2016.03.087. Epub 2016 Apr 21.
  PUBMED    27117410
REFERENCE   2  (bases 1 to 4617)
  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"; doi: 
            "10.1016/j.celrep.2016.03.087"; journalName: "Cell Rep"; date: 
            "2016-05-3- 3"; volume: "15"; issue: "5"; pages: "1100-10"
COMMENT     Sequence Label: env8 16nt J1/13
FEATURES             Location/Qualifiers
     source          1..4617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             187..903
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(217..245)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     840..860
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1244..1262)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1330..1434
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1435..1503,1504..2295)
                     /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(1653..1672)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2466..3054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2955..2974
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3208..3225
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3240..3380
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3482..3673)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      4296..4382
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4474..4501
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcctaaaag cgtagtggga aagtgacgtg aaattcgtcc agattacttg atacggttat
       61 actccgaatg ccacctaggc catacaacat acaacatgag caaggagact caacatgctg
      121 atgattacgc caagcttgca tgcctgcagg tcgactctag aggatccccg ggtaccggta
      181 gaaaaaatga gtaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta
      241 gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg tgatgcaaca
      301 tacggaaaac ttacccttaa atttatttgc actactggaa aactacctgt tccatggcca
      361 acacttgtca ctactttctc ttatggtgtt caatgctttt cccgttatcc ggatcatatg
      421 aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga acgcactata
      481 tctttcaaag atgacgggaa ctacaagacg cgtgctgaag tcaagtttga aggtgatacc
      541 cttgttaatc gtatcgagtt aaaaggtatt gattttaaag aagatggaaa cattctcgga
      601 cacaaactcg agtacaacta taactcacac aatgtataca tcacggcaga caaacaaaag
      661 aatggaatca aagctaactt caaaattcgc cacaacattg aagatggatc cgttcaacta
      721 gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac
      781 cattacctgt cgacacaatc tgccctttcg aaagatccca acgaaaagcg tgaccacatg
      841 gtccttcttg agtttgtaac tgctgctggg attacacatg gcatggatga gctctacaaa
      901 taatgaattc caactgagcg ccggtcgcta ccattaccaa cttgtctggt gtcaaaaata
      961 ataggcctac tagtcggccg tacgggccct ttcgtctcgc gcgtttcggt gatgacggtg
     1021 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     1081 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     1141 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     1201 acagatgcgt aaggagaaaa taccgcatca ggcggcctta agggcctcgt gatacgccta
     1261 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     1321 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1381 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1441 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1501 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1561 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1621 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     1681 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1741 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1801 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1861 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1921 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     1981 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2041 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2101 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2161 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2221 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2281 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2341 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2401 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2461 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2521 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2581 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     2641 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2701 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2761 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2821 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2881 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2941 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3001 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3061 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3121 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3181 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3241 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     3301 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     3361 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3421 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3481 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     3541 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     3601 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     3661 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     3721 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     3781 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     3841 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     3901 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     3961 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     4021 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     4081 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     4141 gccaggaccc aacgctgccc gagaatgcgc attgttgcct ggcggcagta gcgcggtggt
     4201 cccacctgac cccatgccga actcagaagc gaaacgccgt agcgccgatg gtagtgtggg
     4261 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     4321 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     4381 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     4441 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     4501 ttgcgctagc cacagctaac accacgtcgt ccctatctgc tgccctaggt ctatgagtgg
     4561 ttgctggata actttacggg catgcataag gctcgtataa tatattcata taataat
//
