LOCUS       sequence-192116-        4626 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 4626)
  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 4626)
  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 25nt J1/13
FEATURES             Location/Qualifiers
     source          1..4626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             196..912
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(226..254)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     849..869
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1253..1271)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1339..1443
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1444..1512,1513..2304)
                     /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(1662..1681)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2475..3063
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2964..2983
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3217..3234
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3249..3389
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3491..3682)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      4305..4391
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4483..4510
                     /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 acatacgagc aaggagactc
      121 aacatgctga tgattacgcc aagcttgcat gcctgcaggt cgactctaga ggatccccgg
      181 gtaccggtag aaaaaatgag taaaggagaa gaacttttca ctggagttgt cccaattctt
      241 gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt
      301 gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt
      361 ccatggccaa cacttgtcac tactttctct tatggtgttc aatgcttttc ccgttatccg
      421 gatcatatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa
      481 cgcactatat ctttcaaaga tgacgggaac tacaagacgc gtgctgaagt caagtttgaa
      541 ggtgataccc ttgttaatcg tatcgagtta aaaggtattg attttaaaga agatggaaac
      601 attctcggac acaaactcga gtacaactat aactcacaca atgtatacat cacggcagac
      661 aaacaaaaga atggaatcaa agctaacttc aaaattcgcc acaacattga agatggatcc
      721 gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta
      781 ccagacaacc attacctgtc gacacaatct gccctttcga aagatcccaa cgaaaagcgt
      841 gaccacatgg tccttcttga gtttgtaact gctgctggga ttacacatgg catggatgag
      901 ctctacaaat aatgaattcc aactgagcgc cggtcgctac cattaccaac ttgtctggtg
      961 tcaaaaataa taggcctact agtcggccgt acgggccctt tcgtctcgcg cgtttcggtg
     1021 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     1081 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     1141 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     1201 aaataccgca cagatgcgta aggagaaaat accgcatcag gcggccttaa gggcctcgtg
     1261 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     1321 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1381 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     1441 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     1501 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1561 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     1621 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     1681 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     1741 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     1801 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     1861 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     1921 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     1981 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2041 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2101 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2161 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2221 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     2281 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     2341 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2401 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2461 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2521 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2581 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     2641 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2701 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2761 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2821 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2881 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2941 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3001 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3061 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3121 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3181 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3241 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3301 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     3361 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     3421 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3481 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     3541 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     3601 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     3661 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     3721 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     3781 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     3841 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     3901 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     3961 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     4021 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     4081 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
     4141 gcacccgtgg ccaggaccca acgctgcccg agaatgcgca ttgttgcctg gcggcagtag
     4201 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagcg aaacgccgta gcgccgatgg
     4261 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
     4321 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     4381 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
     4441 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg
     4501 atggcctttt tgcgctagcc acagctaaca ccacgtcgtc cctatctgct gccctaggtc
     4561 tatgagtggt tgctggataa ctttacgggc atgcataagg ctcgtataat atattcatat
     4621 aataat
//
