LOCUS       sequence-201363-        2865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the ""Top"" transcript of the Split-Broccoli system under 
            the control of the E. coli sigma 70 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2865)
  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 2865)
  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
FEATURES             Location/Qualifiers
     source          1..2865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    105..126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(141..146,147..164,165..171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..195
                     /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     184..206
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     203..219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     203..219
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      399..445
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(468..485)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(468..484)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(477..499)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(693..712)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     812..834
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(872..890)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        958..1062
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1063..1131,1132..1923)
                     /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(1281..1300)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2094..2682
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2583..2602
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2836..2853
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      121 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      181 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca
      241 tgcctgcagg tcgactctag acgatcccgc gaaattgagc taacaccgtg cgtgttgaca
      301 attttacctc tggcggtgat aatggttgca ggatgatgga gacggtcggg tccaggatca
      361 ttcatggcaa gagacggtcg ggtccagatg atgcggatta gcataacccc ttggggcctc
      421 taaacgggtc ttgaggggtt ttttgcccgg gtaccgagct cgaattcact ggccgtcgtt
      481 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
      541 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
      601 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
      661 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
      721 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
      781 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
      841 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
      901 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
      961 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1021 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1081 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1141 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1201 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1261 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     1321 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1381 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1441 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1501 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     1561 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     1621 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     1681 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     1741 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     1801 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     1861 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     1921 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     1981 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2041 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2101 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2161 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2221 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2281 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2341 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2401 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2461 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2521 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2581 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2641 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2701 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2761 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     2821 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagag
//
