LOCUS       sequence-269475-        3710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7-transcription template for a P2-deletion mutation of the Tte 
            preQ1- riboswitch.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3710)
  AUTHORS   Lund PE, Chatterjee S, Daher M, Walter NG
  TITLE     Protein unties the pseudoknot: S1-mediated unfolding of RNA higher 
            order structure.
  JOURNAL   Nucleic Acids Res. 2019 Dec 13. pii: 5674991. doi: 
            10.1093/nar/gkz1166.
  PUBMED    31832686
REFERENCE   2  (bases 1 to 3710)
  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: "Nucleic 
            Acids Res. 2019 Dec 13. pii: 5674991. doi: 10.1093/nar/gkz1166."
COMMENT     Sequence Label: pUC19_TTE1564_UUCG-loop
FEATURES             Location/Qualifiers
     source          1..3710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1067..1083)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1067..1083)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1080..1102)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1091..1107
                     /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(1115..1121,1122..1139,1140..1145))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1160..1181
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1298..1315)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1469..2057)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1549..1568)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2228..3019,3020..3088))
                     /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     2851..2870
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3089..3193)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3261..3279
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3317..3339)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3439..3458
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3652..3674
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3666..3683
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3667..3683
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cagtgagcaa caaaatgctc acctgggttc gcccagttaa
       61 caaaacaagg gaggtaattt tgtgcccaaa aaaagaataa aagatttagc tgaaattgct
      121 cttgttgcag caatttattt cgcactcaca attatatttt cgtccatttc gtttttaccc
      181 gttcaatttc gaatcgggga aattacgaaa tccattgtag tattcaataa aaaatatgct
      241 atttccatga tgataggaaa tttttttgca aatttgttta gcccatttgc tggtgcaatg
      301 gaattaattt ttatgcctct ttcgaactta ataggctgta caattggata ctacattgga
      361 agacttactc acaaagcgat aggagctata ttcatagccc tttggattgc agcatcagtt
      421 gcaattactt taaaggtttc tgcaggcata ccatttattc cgactttctt aagcgtggga
      481 gtagcggaaa ctgtactttt ggtaactgga tattttttgc ttttcacaat tgaaaagaaa
      541 ggagttgtga aatttgacag ataaatataa agagagaaga tttgacattt acggttacga
      601 aaaaattgac aaagaagttc tagaatctat tgaatatgag tatcctgaaa aaaatactat
      661 cgtggagtat attaccgatg aattttcttc tgtttgccct tggacaggat tacctgacaa
      721 tgcaaaactt actataaggt atatacccca caaaaaactt gtagaactta aatccttaaa
      781 atattacctt acatcttata ggaatgtagg tatattgcaa gaacatgcaa taaacagaat
      841 tttagatgat ttggtggaat tcctgcagcc aaaatttatg gaaataatag gcgaatttca
      901 ggaaagagga ggaatagcta caagaattat agcaaggtat gaaaaagagg agtattaaac
      961 ttaaaaggct gcctaaaatt ttgtaggcag cttttttatt cattttagtt tttcttcaaa
     1021 atgagtttac agagaacgat gaggaagcaa gcttggcgta atcatggtca tagctgtttc
     1081 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     1141 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     1201 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     1261 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     1321 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     1381 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     1441 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1501 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1561 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1621 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1681 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1741 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1801 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1861 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1921 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1981 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2041 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2101 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2161 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2221 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2281 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     2341 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     2401 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     2461 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2521 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2581 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     2641 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     2701 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     2761 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     2821 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     2881 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     2941 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3001 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3061 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3121 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3181 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     3241 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     3301 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     3361 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     3421 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     3481 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     3541 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     3601 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     3661 gacgttgtaa aacgacggcc agtgaattcg agctcggtac ccggggatcc
//
