LOCUS       sequence-378299-        3340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a 0.8 kb bacterial transcription unit with Broccoli-tRNA 
            fluorogenic aptamer shortly upstream of the intrinsic terminator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3340)
  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 3340)
  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: pOP004
FEATURES             Location/Qualifiers
     source          1..3340
                     /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(1105..1122)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      1159..1202
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1259..1847)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1339..1358)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2018..2809,2810..2878))
                     /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     2641..2660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2879..2942)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(3327..3340,1..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 gagtagacac gcattcgtga
      241 agatagctgt agacgtccgc aggttctgga tgttccgtat ctgctgagca ttcagctgga
      301 tagctctcag aagttcattg aacaggatcc tgaaggtcag tatggtctgg aagcagcact
      361 tcgtagcgtg tctctgattc agagctatag tggtagtagc gaactgcagt atgttagcta
      421 tcgtctgagt gaaccagtta gtgatgttca agaatgtcag attcgtggtg ttactcatag
      481 ctcactgctg agtgttgaac tacgtctggt tacctatgaa cgtgaagcac tggaagtcac
      541 gtctcaagat cttcgtgaac aagaggttca tctggtcgaa gttcagctga tgaccgatga
      601 tggcaccagc gttagcagct gcaccgaacg tgttcctgtg agccagctgc atcgtagtcc
      661 aggtgatctg tctgatagtg atgaaggtcg tacccatagc agcagtacag ttctgtagga
      721 tgcacgcatc gttccgtatc gtggtagctg gctggatctg gaacttgatc cgatcaattg
      781 gcgcccggat agctcagtcg gtagagcagc ggagacggtc gggtccagat attcgtatct
      841 gtcgagtaga gtgtgggctc cgcgggtcca gggttcaagt ccctgttcgg gcgccacctg
      901 tacaagatga cctgtaagct taagcccccg gaagatcatc ttccgggggc tttttttttg
      961 ggttgagctc acggaactta tctcgagctg ggcctcatgg gccttccgcc tatcactgcc
     1021 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa catggtcata gctgtttcct
     1081 tgcgtattgg gcgctctccg cttcctcgct cactgactcg ctgcgctcgg tcgttcgggt
     1141 aaagcctggg gtgcctaaat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt
     1201 attgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1261 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1321 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1381 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1441 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1501 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1561 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1621 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1681 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1741 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1801 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1861 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1921 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     1981 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2041 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2101 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2161 aaccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2221 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2281 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2341 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2401 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2461 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2521 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2581 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2641 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2701 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     2761 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     2821 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     2881 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     2941 tatttgaagc taataacagg cctgctggta atcgcaggcc tttttatttg gtgtatttag
     3001 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     3061 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3121 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3181 gtggccgcta cagggcgctc ccattcgcca ttcaggctgc gcaactgttg ggaagggcgt
     3241 ttcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     3301 attaagttgg gtaacgccag ggttttccca gtcacgacgt
//
