LOCUS       sequence-167505-        2882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pVR2 but with an insertion of 1 U immediately following the 
            riboswitch aptamer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2882)
  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 2882)
  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-m5
FEATURES             Location/Qualifiers
     source          1..2882
                     /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(661..677)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(661..677)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(674..696)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    685..701
                     /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(709..715,716..733,734..739))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    754..775
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(892..909)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1063..1651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1143..1162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1822..2613,2614..2682))
                     /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     2445..2464
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2683..2787)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2855..2873
                     /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 aatggacgag ctgtatcctt ggatacggcc ttttttatgt ttttctagag caccttccga
      601 aaaaaggtgt ttttttgcgt gaattagctg tagcgatgtc tctcgcgaat tcgtaatcat
      661 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
      721 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
      781 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
      841 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
      901 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
      961 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1021 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1081 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1141 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1201 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1261 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1321 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1381 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1441 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1501 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1561 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1621 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1681 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     1741 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1801 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1861 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1921 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     1981 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2041 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2101 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     2161 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     2221 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     2281 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     2341 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     2401 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     2461 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     2521 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     2581 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     2641 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     2701 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     2761 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     2821 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     2881 tc
//
