LOCUS       sequence-201364-        2731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the ""Bottom"" transcript of the Split-Broccoli system 
            under the control of the T7 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2731)
  AUTHORS   Alam KK, Tawiah KD, Lichte MF, Porciani D, Burke DH
  TITLE     A Fluorescent Split Aptamer for Visualizing RNA-RNA Assembly In 
            Vivo.
  JOURNAL   ACS Synth Biol. 2017 May 26. doi: 10.1021/acssynbio.7b00059.
  PUBMED    28548488
REFERENCE   2  (bases 1 to 2731)
  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: "ACS Synth 
            Biol. 2017 May 26. doi: 10.1021/acssynbio.7b00059."
COMMENT     Sequence Label: pUC19-T7-Bottom-T
FEATURES             Location/Qualifiers
     source          1..2731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(45..64)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     164..186
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(224..242)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        310..414
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(415..483,484..1275)
                     /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(633..652)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1446..2034
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1935..1954
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2188..2205
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2322..2342
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2359..2377
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2481..2528
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2551..2568)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2551..2567)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2560..2582)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
       61 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
      121 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
      181 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
      241 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
      301 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      361 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      421 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      481 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      541 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      601 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      661 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
      721 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
      781 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
      841 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
      901 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
      961 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1021 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1081 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1141 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1201 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1261 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1321 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1381 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1441 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1501 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1561 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1621 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1681 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1741 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     1801 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     1861 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     1921 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     1981 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2041 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2101 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2161 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2221 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2281 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     2341 cagacgatcc cgcgaaatta atacgactca ctataggatc cgcatcatct gtcgagtaga
     2401 gtgtgggctc ttgccatgtg tatgtgggtc aacccacata ctctgatgat cctgtcgagt
     2461 agagtgtggg ctccatcatc ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     2521 gttttttgcc cgggtaccga gctcgaattc actggccgtc gttttacaac gtcgtgactg
     2581 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     2641 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     2701 cgaatggcgc ctgatgcggt attttctcct t
//
