LOCUS       sequence-192119-        4608 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 4608)
  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 4608)
  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 reverse J1/13
FEATURES             Location/Qualifiers
     source          1..4608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             178..894
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(208..236)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     831..851
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1235..1253)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1321..1425
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1426..1494,1495..2286)
                     /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(1644..1663)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2457..3045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2946..2965
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3199..3216
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3231..3371
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3473..3664)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      4287..4373
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4465..4492
                     /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 ccaacataga gcaaggagac tcaacatgct gatgattacg
      121 ccaagcttgc atgcctgcag gtcgactcta gaggatcccc gggtaccggt agaaaaaatg
      181 agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat
      241 gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa
      301 cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc
      361 actactttct cttatggtgt tcaatgcttt tcccgttatc cggatcatat gaaacggcat
      421 gactttttca agagtgccat gcccgaaggt tatgtacagg aacgcactat atctttcaaa
      481 gatgacggga actacaagac gcgtgctgaa gtcaagtttg aaggtgatac ccttgttaat
      541 cgtatcgagt taaaaggtat tgattttaaa gaagatggaa acattctcgg acacaaactc
      601 gagtacaact ataactcaca caatgtatac atcacggcag acaaacaaaa gaatggaatc
      661 aaagctaact tcaaaattcg ccacaacatt gaagatggat ccgttcaact agcagaccat
      721 tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg
      781 tcgacacaat ctgccctttc gaaagatccc aacgaaaagc gtgaccacat ggtccttctt
      841 gagtttgtaa ctgctgctgg gattacacat ggcatggatg agctctacaa ataatgaatt
      901 ccaactgagc gccggtcgct accattacca acttgtctgg tgtcaaaaat aataggccta
      961 ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     1021 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     1081 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     1141 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     1201 taaggagaaa ataccgcatc aggcggcctt aagggcctcg tgatacgcct atttttatag
     1261 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     1321 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     1381 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     1441 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     1501 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     1561 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     1621 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     1681 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     1741 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     1801 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     1861 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     1921 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     1981 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     2041 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     2101 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     2161 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2221 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     2281 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     2341 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     2401 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2461 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2521 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2581 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     2641 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2701 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2761 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2821 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2881 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2941 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3001 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3061 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3121 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3181 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     3241 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     3301 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     3361 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3421 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3481 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     3541 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     3601 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     3661 tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac
     3721 gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt
     3781 gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc
     3841 aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg
     3901 cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg
     3961 aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc
     4021 agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc
     4081 gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggccaggacc
     4141 caacgctgcc cgagaatgcg cattgttgcc tggcggcagt agcgcggtgg tcccacctga
     4201 ccccatgccg aactcagaag cgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca
     4261 tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     4321 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
     4381 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
     4441 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgctag
     4501 ccacagctaa caccacgtcg tccctatctg ctgccctagg tctatgagtg gttgctggat
     4561 aactttacgg gcatgcataa ggctcgtata atatattcat ataataat
//
