LOCUS       sequence-192689-        2810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ARiBo-fused RNA in vitro with T7 RNA polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2810)
  AUTHORS   Di Tomasso G, Lampron P, Dagenais P, Omichinski JG, Legault P
  TITLE     The ARiBo tag: a reliable tool for affinity purification of RNAs 
            under native conditions.
  JOURNAL   Nucleic Acids Res. 2011 Feb;39(3):e18. doi: 10.1093/nar/gkq1084. 
            Epub 2010 Nov 11.
  PUBMED    21071425
REFERENCE   2  (bases 1 to 2810)
  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"; journalName: "Nucleic 
            Acids Res."; date: "2011-02"; volume: "39(3):e18. doi"; pages: " 
            10.1093/nar/gkq1084. Epub 2010 Nov 11"
COMMENT     Sequence Label: pARiBo1
FEATURES             Location/Qualifiers
     source          1..2810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     302..321
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     555..572
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    689..710
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(725..730,731..748,749..755)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    763..779
                     /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)."
     primer_bind     768..790
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     787..803
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     787..803
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        822..840
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    857..871
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     primer_bind     complement(997..1014)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(997..1013)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1006..1028)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1222..1241)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1341..1363
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1401..1419)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1487..1591
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1592..1660,1661..2452)
                     /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     complement(1810..1829)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2623..2810,1..401)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
       61 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      121 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      181 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
      241 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
      301 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
      361 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
      421 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
      481 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
      541 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
      601 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
      661 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
      721 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
      781 ttcacacagg aaacagctat gaccatgatt acgccaagct ttaatacgac tcactatagg
      841 cgaagcgccg aactgggccc tgaagaaggg ctcagttgac gaggtggggt ttatcgagat
      901 ttcggcggat gactcccggt tgttcatcac aaccgcaaac ttttacttaa atcattaagg
      961 tgacttagtg gacaaaggtg aaagtgtgat gaattcactg gccgtcgttt tacaacgtcg
     1021 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     1081 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     1141 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     1201 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     1261 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     1321 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     1381 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     1441 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1501 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     1561 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     1621 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     1681 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     1741 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1801 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     1861 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     1921 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     1981 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2041 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2101 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     2161 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2221 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2281 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2341 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2401 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     2461 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     2521 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2581 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2641 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2701 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2761 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
//
