LOCUS       sequence-192121-        4625 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 4625)
  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 4625)
  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 24nt J1/13
FEATURES             Location/Qualifiers
     source          1..4625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             195..911
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(225..253)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     848..868
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1252..1270)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1338..1442
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1443..1511,1512..2303)
                     /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(1661..1680)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2474..3062
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2963..2982
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3216..3233
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3248..3388
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3490..3681)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      4304..4390
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4482..4509
                     /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 acaacataca acatagagca aggagactca
      121 acatgctgat gattacgcca agcttgcatg cctgcaggtc gactctagag gatccccggg
      181 taccggtaga aaaaatgagt aaaggagaag aacttttcac tggagttgtc ccaattcttg
      241 ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg
      301 atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc
      361 catggccaac acttgtcact actttctctt atggtgttca atgcttttcc cgttatccgg
      421 atcatatgaa acggcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaac
      481 gcactatatc tttcaaagat gacgggaact acaagacgcg tgctgaagtc aagtttgaag
      541 gtgataccct tgttaatcgt atcgagttaa aaggtattga ttttaaagaa gatggaaaca
      601 ttctcggaca caaactcgag tacaactata actcacacaa tgtatacatc acggcagaca
      661 aacaaaagaa tggaatcaaa gctaacttca aaattcgcca caacattgaa gatggatccg
      721 ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac
      781 cagacaacca ttacctgtcg acacaatctg ccctttcgaa agatcccaac gaaaagcgtg
      841 accacatggt ccttcttgag tttgtaactg ctgctgggat tacacatggc atggatgagc
      901 tctacaaata atgaattcca actgagcgcc ggtcgctacc attaccaact tgtctggtgt
      961 caaaaataat aggcctacta gtcggccgta cgggcccttt cgtctcgcgc gtttcggtga
     1021 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     1081 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     1141 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     1201 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cggccttaag ggcctcgtga
     1261 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     1321 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1381 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1441 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1501 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1561 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1621 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1681 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     1741 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     1801 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1861 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1921 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1981 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     2041 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     2101 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2161 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2221 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2281 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     2341 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2401 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2461 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2521 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2581 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     2641 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2701 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2761 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2821 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2881 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2941 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3001 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3061 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3121 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3181 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3241 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     3301 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     3361 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     3421 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3481 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     3541 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     3601 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     3661 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     3721 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     3781 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     3841 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     3901 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     3961 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     4021 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     4081 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     4141 cacccgtggc caggacccaa cgctgcccga gaatgcgcat tgttgcctgg cggcagtagc
     4201 gcggtggtcc cacctgaccc catgccgaac tcagaagcga aacgccgtag cgccgatggt
     4261 agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc
     4321 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     4381 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
     4441 ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga
     4501 tggccttttt gcgctagcca cagctaacac cacgtcgtcc ctatctgctg ccctaggtct
     4561 atgagtggtt gctggataac tttacgggca tgcataaggc tcgtataata tattcatata
     4621 ataat
//
