LOCUS       sequence-167518-        2881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pVR2 but with two G to U mutations immediately following the 
            riboswitch aptamer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2881)
  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 2881)
  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-m8
FEATURES             Location/Qualifiers
     source          1..2881
                     /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(660..676)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(660..676)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(673..695)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    684..700
                     /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(708..714,715..732,733..738))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    753..774
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(891..908)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1062..1650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1142..1161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1821..2612,2613..2681))
                     /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     2444..2463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2682..2786)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2854..2872
                     /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 gtcccaaaac acttgatact gtatgagcat acagtataat tgcttcggta
      481 ccgctctgcg ttccgtgcga tatgcgggag aggttctagc tacaccctct ataaaaaact
      541 aattacgagc tgtatccttg gatacggcct tttttatgtt tttctagagc accttccgaa
      601 aaaaggtgtt tttttgcgtg aattagctgt agcgatgtct ctcgcgaatt cgtaatcatg
      661 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
      721 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
      781 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
      841 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
      901 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
      961 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1021 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1081 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1141 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1201 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1261 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1321 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1381 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1441 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1501 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1561 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1621 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1681 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1741 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     1801 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1861 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1921 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1981 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2041 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2101 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2161 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2221 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2281 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     2341 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2401 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     2461 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2521 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2581 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2641 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2701 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2761 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     2821 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     2881 c
//
