LOCUS       sequence-192122-        4618 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 4618)
  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 4618)
  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 17nt J1/13
FEATURES             Location/Qualifiers
     source          1..4618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             188..904
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(218..246)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     841..861
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1245..1263)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1331..1435
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1436..1504,1505..2296)
                     /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(1654..1673)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2467..3055
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2956..2975
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3209..3226
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3241..3381
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3483..3674)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      4297..4383
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4475..4502
                     /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 acaacataga gcaaggagac tcaacatgct
      121 gatgattacg ccaagcttgc atgcctgcag gtcgactcta gaggatcccc gggtaccggt
      181 agaaaaaatg agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt
      241 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac
      301 atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc
      361 aacacttgtc actactttct cttatggtgt tcaatgcttt tcccgttatc cggatcatat
      421 gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg aacgcactat
      481 atctttcaaa gatgacggga actacaagac gcgtgctgaa gtcaagtttg aaggtgatac
      541 ccttgttaat cgtatcgagt taaaaggtat tgattttaaa gaagatggaa acattctcgg
      601 acacaaactc gagtacaact ataactcaca caatgtatac atcacggcag acaaacaaaa
      661 gaatggaatc aaagctaact tcaaaattcg ccacaacatt gaagatggat ccgttcaact
      721 agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa
      781 ccattacctg tcgacacaat ctgccctttc gaaagatccc aacgaaaagc gtgaccacat
      841 ggtccttctt gagtttgtaa ctgctgctgg gattacacat ggcatggatg agctctacaa
      901 ataatgaatt ccaactgagc gccggtcgct accattacca acttgtctgg tgtcaaaaat
      961 aataggccta ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg tgatgacggt
     1021 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     1081 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     1141 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     1201 cacagatgcg taaggagaaa ataccgcatc aggcggcctt aagggcctcg tgatacgcct
     1261 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     1321 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1381 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1441 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1501 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1561 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1621 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     1681 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1741 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1801 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1861 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1921 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     1981 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2041 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2101 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2161 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2221 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2281 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2341 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2401 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2461 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2521 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2581 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     2641 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2701 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2761 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2821 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2881 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2941 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3001 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3061 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3121 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3181 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3241 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     3301 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     3361 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3421 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3481 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     3541 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     3601 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     3661 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     3721 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     3781 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     3841 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     3901 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
     3961 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     4021 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
     4081 aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt
     4141 ggccaggacc caacgctgcc cgagaatgcg cattgttgcc tggcggcagt agcgcggtgg
     4201 tcccacctga ccccatgccg aactcagaag cgaaacgccg tagcgccgat ggtagtgtgg
     4261 ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg
     4321 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     4381 aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga
     4441 cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt
     4501 tttgcgctag ccacagctaa caccacgtcg tccctatctg ctgccctagg tctatgagtg
     4561 gttgctggat aactttacgg gcatgcataa ggctcgtata atatattcat ataataat
//
