LOCUS       sequence-292718-        3443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for Malachite Green Aptamer (MGA) transcription assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3443)
  AUTHORS   Newing TP, Oakley AJ, Miller M, Dawson CJ, Brown SHJ, Bouwer JC, 
            Tolun G, Lewis PJ
  TITLE     Molecular basis for RNA polymerase-dependent transcription complex 
            recycling by the helicase-like motor protein HelD.
  JOURNAL   Nat Commun. 2020 Dec 18;11(1):6420. doi: 10.1038/s41467-020-20157-5.
  PUBMED    33339820
REFERENCE   2  (bases 1 to 3443)
  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: "Nat 
            Commun."; date: "2020-12-18"; volume: "11(1):6420. doi"; pages: " 
            10.1038/s41467-020-20157-5"
COMMENT     Sequence Label: pNG1299
FEATURES             Location/Qualifiers
     source          1..3443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(538..543,544..561,562..568)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     terminator      1112..1155
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(1222..1238)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1222..1238)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1235..1257)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1246..1262
                     /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)."
     promoter        complement(join(1270..1276,1277..1294,1295..1300))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1315..1336
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1453..1470)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1624..2212)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1704..1723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2383..3174,3175..3243))
                     /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     3006..3025
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3244..3348)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3416..3434
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt ggagatcggt acttcgcgaa
      421 tgcgtcgaga tatcctgcag ttgacattcc ccccgccccg gggtaccctc cttcccggga
      481 ggaggagaac tcgagttgac aaattgcagt aggcatgaca aaatggactc atgtacattt
      541 acactttatg cttccggctc gtataatgac tagtggatcc cgactggcga gagccaggta
      601 acgaatggat cctaaaaacg gatcccgact ggcgagagcc aggtaacgaa tggatcctaa
      661 aaacggatcc cgactggcga gagccaggta acgaatggat cctaaaaacg gatcccgact
      721 ggcgagagcc aggtaacgaa tggatcctaa aaacggatcc cgactggcga gagccaggta
      781 acgaatggat cctaaaaacg gatcccgact ggcgagagcc aggtaacgaa tggatcctaa
      841 aaacggatcc cgactggcga gagccaggta acgaatggat cctaaaaacg gatcccgact
      901 ggcgagagcc aggtaacgaa tggatcctaa aaacggatcc cgactggcga gagccaggta
      961 acgaatggat cctaaaaacg gatcccgact ggcgagagcc aggtaacgaa tggatcctaa
     1021 aaacggatcc cgactggcga gagccaggta acgaatggat cctaaaaacg gatcccgact
     1081 ggcgagagcc aggtaacgaa tggatcccat ggaaatcatc cttagcgaaa gctaaggatt
     1141 ttttttatct gaaataccgg tgatatcgga tgccgggacc gacgagtgca gaggcgtgca
     1201 agcgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     1261 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     1321 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1381 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1441 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1501 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1561 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1621 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1681 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1741 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1801 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1861 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1921 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1981 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2041 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2101 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2161 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2221 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2281 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2341 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2401 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2461 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2521 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2581 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2641 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2701 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2761 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2821 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2881 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     2941 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3001 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3061 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3121 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3181 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3241 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3301 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3361 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     3421 gcgtatcacg aggccctttc gtc
//
