LOCUS       sequence-174671-        2968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the ""Bottom"" transcript of the Split-Broccoli system 
            downstream of an RNA Toehold switch Trigger sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2968)
  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 2968)
  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-Trigger-Bottom
FEATURES             Location/Qualifiers
     source          1..2968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     8..25
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    142..163
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(178..183,184..201,202..208)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    216..232
                     /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     221..243
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     240..256
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     240..256
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(299..346)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(608..625)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(608..624)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(617..639)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(833..852)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     952..974
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1012..1030)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1098..1202
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1203..1271,1272..2063)
                     /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(1421..1440)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2234..2822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2723..2742
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
       61 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
      121 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
      181 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
      241 aggaaacagc tatgaccatg attacgccaa gcttgcatgc ctgcaggtcg actctagaca
      301 aaaaacccct caagacccgt ttagaggccc caaggggtta tgctaggatg atggagccca
      361 cactctactc gacaggatca tcagagtatg tgggttgacc cacatacaca tggcaagagc
      421 ccacactcta ctcgacagat gatgcggatc cctatcttat cttatctatc tcgtttatcc
      481 ctgctttact gactattgca cagaatagtc agtccctgca accattatca ccgccagagg
      541 taaaattgtc aacacgcacg gtgttagctc aatttcgcgg gatcgcccgg gtaccgagct
      601 cgaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
      661 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
      721 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt
      781 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
      841 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
      901 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
      961 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     1021 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     1081 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1141 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1201 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1261 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1321 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1381 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1441 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     1501 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     1561 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1621 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1681 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1741 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     1801 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     1861 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     1921 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     1981 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2041 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     2101 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2161 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2221 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2281 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2341 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2401 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2461 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2521 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2581 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2641 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2701 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2761 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2821 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2881 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     2941 ctgataccgc tcgccgcagc cgaacgac
//
