LOCUS       sequence-371540-        3252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a 0.8 kb bacterial transcription unit with Broccoli-tRNA 
            fluorogenic aptamer shortly downstream of the promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3252)
  AUTHORS   Huang YH, Trapp V, Puro O, Makinen JJ, Metsa-Ketela M, Wahl MC, 
            Belogurov GA
  TITLE     Fluorogenic RNA aptamers to probe transcription initiation and 
            co-transcriptional RNA folding by multi-subunit RNA polymerases.
  JOURNAL   Methods Enzymol. 2022;675:207-233. doi: 10.1016/bs.mie.2022.07.010. 
            Epub 2022 Aug 19.
  PUBMED    36220271
REFERENCE   2  (bases 1 to 3252)
  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: "Methods 
            Enzymol."; date: "2022"; volume: "675:207-233. doi"; pages: " 
            10.1016/bs.mie.2022.07.010. Epub 2022 Aug 19"
COMMENT     Sequence Label: pOP005
FEATURES             Location/Qualifiers
     source          1..3252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..18
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2..18
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     29..48
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        29..47
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1017..1034)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      1071..1114
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1171..1759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1251..1270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1930..2721,2722..2790))
                     /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     2553..2572
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2791..2854)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(3239..3252,1..9)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tgtaaaacga cggccagtga gcgcgacgta atacgactca ctatagggcg aattgtaggc
       61 ggaaggccgt caaggccgca ttctagagga gagacaactt aaagagacat tctgtgcgca
      121 taccatccct gcctgcgttg acgaaactcg ttaacatgtt ataatacaag catcgagagg
      181 gacacgagga accagaggag ttctggatca tagctagcac gcccggatag ctcagtcggt
      241 agagcagcgg agacggtcgg gtccagatat tcgtatctgt cgagtagagt gtgggctccg
      301 cgggtccagg gttcaagtcc ctgttcgggc gccacaggat cctgaaggtc agtatggtct
      361 ggaagcagca cttcgtagcg tgtctctgat tcagagctat agtggtagta gcgaactgca
      421 gtatgttagc tatcgtctga gtgaaccagt tagtgatgtt caagaatgtc agattcgtgg
      481 tgttactcat agctcactgc tgagtgttga actacgtctg gttacctatg aacgtgaagc
      541 actggaagtc acgtctcaag atcttcgtga acaagaggtt catctggtcg aagttcagct
      601 gatgaccgat gatggcacca gcgttagcag ctgcaccgaa cgtgttcctg tgagccagct
      661 gcatcgtagt ccaggtgatc tgtctgatag tgatgaaggt cgtacccata gcagcagtac
      721 agttctgtag gatgcacgca tcgttccgta tcgtggtagc tggctggatc tggaacttga
      781 tccgatcaat tggcgaattt tttaagtgcc tgtacaagat gacctgtaag cttaagcccc
      841 cggaagatca tcttccgggg gctttttttt tgggttgagc tcacggaact tatctcgagc
      901 tgggcctcat gggccttccg cctatcactg cccgctttcc agtcgggaaa cctgtcgtgc
      961 cagctgcatt aacatggtca tagctgtttc cttgcgtatt gggcgctctc cgcttcctcg
     1021 ctcactgact cgctgcgctc ggtcgttcgg gtaaagcctg gggtgcctaa ataaaacgaa
     1081 aggcccagtc ttccgactga gcctttcgtt ttattgagca aaaggccagc aaaaggccag
     1141 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1201 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1261 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1321 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1381 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1441 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1501 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1561 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1621 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1681 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1741 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1801 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1861 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1921 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1981 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2041 atctggcccc agtgctgcaa tgataccgcg agaaccacgc tcaccggctc cagatttatc
     2101 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2161 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2221 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2281 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2341 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2401 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2461 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     2521 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     2581 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     2641 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     2701 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     2761 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     2821 ttatcagggt tattgtctca tgagcggata catatttgaa gctaataaca ggcctgctgg
     2881 taatcgcagg cctttttatt tggtgtattt agaaaaataa acaaataggg gttccgcgca
     2941 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
     3001 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3061 taaatcaaaa gaatagaccg agatagggtt gagtggccgc tacagggcgc tcccattcgc
     3121 cattcaggct gcgcaactgt tgggaagggc gtttcggtgc gggcctcttc gctattacgc
     3181 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     3241 cagtcacgac gt
//
