LOCUS       sequence-306846-        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"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(182..225)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        complement(join(769..775,776..793,794..799))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     primer_bind     complement(942..959)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(942..958)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(951..973)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1167..1186)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1286..1308
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1346..1364)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1432..1536
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1537..1605,1606..2397)
                     /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(1755..1774)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2568..3156
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3057..3076
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3310..3327
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag ctcgcttgca cgcctctgca ctcgtcggtc ccggcatccg atatcaccgg
      181 tatttcagat aaaaaaaatc cttagctttc gctaaggatg atttccatgg gatccattcg
      241 ttacctggct ctcgccagtc gggatccgtt tttaggatcc attcgttacc tggctctcgc
      301 cagtcgggat ccgtttttag gatccattcg ttacctggct ctcgccagtc gggatccgtt
      361 tttaggatcc attcgttacc tggctctcgc cagtcgggat ccgtttttag gatccattcg
      421 ttacctggct ctcgccagtc gggatccgtt tttaggatcc attcgttacc tggctctcgc
      481 cagtcgggat ccgtttttag gatccattcg ttacctggct ctcgccagtc gggatccgtt
      541 tttaggatcc attcgttacc tggctctcgc cagtcgggat ccgtttttag gatccattcg
      601 ttacctggct ctcgccagtc gggatccgtt tttaggatcc attcgttacc tggctctcgc
      661 cagtcgggat ccgtttttag gatccattcg ttacctggct ctcgccagtc gggatccgtt
      721 tttaggatcc attcgttacc tggctctcgc cagtcgggat ccactagtca ttatacgagc
      781 cggaagcata aagtgtaaat gtacatgagt ccattttgtc atgcctactg caatttgtca
      841 actcgagttc tcctcctccc gggaaggagg gtaccccggg gcggggggaa tgtcaactgc
      901 aggatatctc gacgcattcg cgaagtaccg atctccaatt cactggccgt cgttttacaa
      961 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     1021 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     1081 agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     1141 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     1201 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     1261 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     1321 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     1381 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     1441 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1501 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     1561 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     1621 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     1681 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     1741 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     1801 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     1861 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     1921 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     1981 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2041 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2101 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2161 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2221 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2281 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2341 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     2401 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     2461 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     2521 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2581 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2641 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2701 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2761 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2821 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2881 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2941 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3001 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3061 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3121 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3181 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3241 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3301 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     3361 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     3421 gcgggcagtg agcgcaacgc aat
//
