LOCUS       sequence-174701-        3073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the modular Split-Broccoli system (Top + Bottom) under the
            control of E. coli sigma 70 promoters..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3073)
  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 3073)
  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-P70a-Top-T~P70a-Bottom-T
FEATURES             Location/Qualifiers
     source          1..3073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..22
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    139..160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(175..180,181..198,199..205)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    213..229
                     /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     218..240
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     237..253
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     237..253
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(267..314)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     656..678
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     670..687
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     671..687
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    688..744
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(935..954)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1054..1076
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1114..1132)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1200..1304
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1305..1373,1374..2165)
                     /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(1523..1542)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2336..2924
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2825..2844
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
       61 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      121 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
      181 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
      241 aaacagctat gaccatgatt acgccacaaa aaacccctca agacccgttt agaggcccca
      301 aggggttatg ctaggatgat ggagcccaca ctctactcga caggatcatc agagtatgtg
      361 ggttgaccca catacacatg gcaagagccc acactctact cgacagatga tgcggatcct
      421 gcaaccatta tcaccgccag aggtaaaatt gtcaacacgc acggtgttag ctcacatatg
      481 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
      541 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
      601 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
      661 tcacgacgtt gtaaaacgac ggccagtgaa ttcgagctcg gtacccgggg atcctctaga
      721 gtcgacctgc aggcatgcaa gcttaaaaaa aagcccgcct ttcgggggct ttgatccgca
      781 tcatctggac cagaccgtct cttgccatga atgatcctgg acccgaccgt ctccatcatc
      841 ctgcaaccat tatcaccgcc agaggtaaaa ttgtcaacac gcacggtgtt agctcaattt
      901 cgcgggatcg tctagacgat tggtgcactc tcagtacaat ctgctctgat gccgcatagt
      961 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     1021 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     1081 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     1141 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     1201 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1261 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1321 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1381 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     1441 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     1501 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     1561 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     1621 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     1681 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     1741 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     1801 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     1861 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     1921 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     1981 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2041 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2101 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2161 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2221 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2281 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2341 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2401 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2461 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2521 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2581 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2641 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2701 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2761 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2821 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2881 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2941 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3001 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3061 gccgaacgac cga
//
