LOCUS       sequence-192123-        4620 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 4620)
  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 4620)
  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 19nt J1/13
FEATURES             Location/Qualifiers
     source          1..4620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             190..906
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(220..248)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     843..863
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1247..1265)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1333..1437
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1438..1506,1507..2298)
                     /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(1656..1675)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2469..3057
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2958..2977
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3211..3228
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3243..3383
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3485..3676)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      4299..4385
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4477..4504
                     /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 gagcaaggag actcaacatg
      121 ctgatgatta cgccaagctt gcatgcctgc aggtcgactc tagaggatcc ccgggtaccg
      181 gtagaaaaaa tgagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa
      241 ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca
      301 acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg
      361 ccaacacttg tcactacttt ctcttatggt gttcaatgct tttcccgtta tccggatcat
      421 atgaaacggc atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaacgcact
      481 atatctttca aagatgacgg gaactacaag acgcgtgctg aagtcaagtt tgaaggtgat
      541 acccttgtta atcgtatcga gttaaaaggt attgatttta aagaagatgg aaacattctc
      601 ggacacaaac tcgagtacaa ctataactca cacaatgtat acatcacggc agacaaacaa
      661 aagaatggaa tcaaagctaa cttcaaaatt cgccacaaca ttgaagatgg atccgttcaa
      721 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac
      781 aaccattacc tgtcgacaca atctgccctt tcgaaagatc ccaacgaaaa gcgtgaccac
      841 atggtccttc ttgagtttgt aactgctgct gggattacac atggcatgga tgagctctac
      901 aaataatgaa ttccaactga gcgccggtcg ctaccattac caacttgtct ggtgtcaaaa
      961 ataataggcc tactagtcgg ccgtacgggc cctttcgtct cgcgcgtttc ggtgatgacg
     1021 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     1081 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     1141 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     1201 cgcacagatg cgtaaggaga aaataccgca tcaggcggcc ttaagggcct cgtgatacgc
     1261 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     1321 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     1381 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     1441 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     1501 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     1561 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     1621 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     1681 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     1741 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     1801 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1861 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1921 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1981 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     2041 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2101 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     2161 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2221 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2281 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     2341 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     2401 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2461 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2521 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2581 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     2641 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2701 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2761 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2821 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2881 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2941 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3001 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3061 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3121 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3181 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3241 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     3301 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     3361 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     3421 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     3481 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     3541 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     3601 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     3661 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
     3721 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
     3781 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
     3841 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
     3901 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
     3961 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
     4021 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
     4081 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
     4141 gtggccagga cccaacgctg cccgagaatg cgcattgttg cctggcggca gtagcgcggt
     4201 ggtcccacct gaccccatgc cgaactcaga agcgaaacgc cgtagcgccg atggtagtgt
     4261 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     4321 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     4381 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     4441 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
     4501 tttttgcgct agccacagct aacaccacgt cgtccctatc tgctgcccta ggtctatgag
     4561 tggttgctgg ataactttac gggcatgcat aaggctcgta taatatattc atataataat
//
