LOCUS       sequence-174665-        2860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the fluorescence-activating Stabilized dimeric Broccoli 
            (SdB) aptamer under the control of the T7 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2860)
  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 2860)
  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-SdB-T
FEATURES             Location/Qualifiers
     source          1..2860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     61..78
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    195..216
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(231..236,237..254,255..261)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    269..285
                     /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     274..296
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     293..309
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     293..309
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        366..384
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      483..530
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(553..570)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(553..569)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(562..584)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(778..797)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     897..919
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(957..975)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1043..1147
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1148..1216,1217..2008)
                     /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(1366..1385)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2179..2767
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2668..2687
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
       61 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
      121 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
      181 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
      241 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
      301 agctatgacc atgattacgc caagcttgca tgcctgcagg tcgactctag acgatcccgc
      361 gaaattaata cgactcacta taggagggag acggtcgggt ccatctgaga cggtcgggtc
      421 cagatattcg tatctgtcga gtagagtgtg ggctcagatg tcgagtagag tgtgggctcc
      481 ctctagcata accccttggg gcctctaaac gggtcttgag gggttttttg cccgggtacc
      541 gagctcgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
      601 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
      661 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
      721 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
      781 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
      841 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
      901 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
      961 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     1021 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1081 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1141 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1201 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1261 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1321 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1381 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1441 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1501 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1561 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1621 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1681 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1741 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1801 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1861 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     1921 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     1981 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2041 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2101 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2161 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2221 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2281 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2341 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2401 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2461 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2521 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2581 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2641 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2701 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2761 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2821 tcacatgttc tttcctgcgt tatcccctga ttctgtggat
//
