LOCUS       sequence-167510-        2883 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pVR2 but with an insertion of 2 U's immediately following the 
            riboswitch aptamer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2883)
  AUTHORS   Widom JR, Nedialkov YA, Rai V, Hayes RL, Brooks CL 3rd, Artsimovitch
            I, Walter NG
  TITLE     Ligand Modulates Cross-Coupling between Riboswitch Folding and 
            Transcriptional Pausing.
  JOURNAL   Mol Cell. 2018 Nov 1;72(3):541-552.e6. doi: 
            10.1016/j.molcel.2018.08.046.
  PUBMED    30388413
REFERENCE   2  (bases 1 to 2883)
  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.molcel.2018.08"; journalName: "Mol Cell"; date: 
            "2018-11-1- 1"; volume: "72"; issue: "3"; pages: "541-552.e6"
COMMENT     Sequence Label: pVR2-m6
FEATURES             Location/Qualifiers
     source          1..2883
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(662..678)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(662..678)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(675..697)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    686..702
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(710..716,717..734,735..740))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    755..776
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(893..910)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1064..1652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1144..1163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1823..2614,2615..2683))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2446..2465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2684..2788)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2856..2874
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc ctgcaggtcg
      421 actctagagg atcccaaaac acttgatact gtatgagcat acagtataat tgcttcggta
      481 ccgctctgcg ttccgtgcga tatgcgggag aggttctagc tacaccctct ataaaaaact
      541 aattggacga gctgtatcct tggatacggc cttttttatg tttttctaga gcaccttccg
      601 aaaaaaggtg tttttttgcg tgaattagct gtagcgatgt ctctcgcgaa ttcgtaatca
      661 tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
      721 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
      781 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
      841 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
      901 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
      961 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1021 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1081 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1141 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1201 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1261 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1321 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1381 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     1441 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     1501 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     1561 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     1621 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     1681 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     1741 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     1801 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     1861 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     1921 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     1981 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     2041 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     2101 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     2161 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     2221 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     2281 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     2341 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     2401 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     2461 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     2521 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     2581 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     2641 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     2701 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     2761 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     2821 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     2881 gtc
//
