LOCUS       sequence-192687-        2804 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 2804)
  AUTHORS   Salvail-Lacoste A, Di Tomasso G, Piette BL, Legault P
  TITLE     Affinity purification of T7 RNA transcripts with homogeneous ends 
            using ARiBo and CRISPR tags.
  JOURNAL   RNA. 2013 Jul;19(7):1003-14. doi: 10.1261/rna.037432.112. Epub 2013 
            May 8.
  PUBMED    23657939
REFERENCE   2  (bases 1 to 2804)
  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: 
            "10.1261/rna.037432.112"; journalName: "RNA"; date: "2013-07"; 
            volume: "19"; issue: "7"; pages: "1003-14"
COMMENT     Sequence Label: pARiBo4
FEATURES             Location/Qualifiers
     source          1..2804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     162..179
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    296..317
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(332..337,338..355,356..362)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    370..386
                     /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     375..397
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     394..410
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     394..410
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    458..472
                     /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(598..615)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(598..614)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(607..629)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(823..842)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     942..964
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1002..1020)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1088..1192
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1193..1261,1262..2053)
                     /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(1411..1430)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2224..2804,1..8)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2713..2732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
       61 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
      121 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      181 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      241 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      301 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
      361 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
      421 ccaagcttta atacgactca ctatagcgcc gaactgggcc ctgaagaagg gctcagttga
      481 cgaggtagac gttatcgaga tttcggcgga tgcgtctcgg ttgttcatca caaccgcaaa
      541 cttttactta aatcattaag gtgacttagt ggacaaaggt gaaagtgtga tgaattcact
      601 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
      661 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
      721 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
      781 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
      841 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
      901 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
      961 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     1021 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     1081 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1141 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1201 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1261 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1321 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1381 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1441 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1501 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1561 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1621 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1681 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1741 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     1801 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1861 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     1921 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     1981 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2041 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     2101 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2161 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2221 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2281 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2341 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     2401 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2461 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2521 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2581 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2641 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2701 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2761 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcc
//
