LOCUS       sequence-98514-e        3790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7-transcription template for Tte mRNA used in SiM-KARTS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3790)
  AUTHORS   Rinaldi AJ, Lund PE, Blanco MR, Walter NG
  TITLE     The Shine-Dalgarno sequence of riboswitch-regulated single mRNAs 
            shows ligand-dependent accessibility bursts.
  JOURNAL   Nat Commun. 2016 Jan 19;7:8976. doi: 10.1038/ncomms9976.
  PUBMED    26781350
REFERENCE   2  (bases 1 to 3790)
  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: "2016-01-19"; volume: "7:8976. doi"; pages: " 
            10.1038/ncomms9976"
COMMENT     Sequence Label: pUC19_extendedTTE1564
FEATURES             Location/Qualifiers
     source          1..3790
                     /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"
     primer_bind     423..442
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        423..441
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1569..1585)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1569..1585)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1582..1604)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1593..1609
                     /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(1617..1623,1624..1641,1642..1647))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1662..1683
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1800..1817)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1971..2559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2051..2070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2730..3521,3522..3590))
                     /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     3353..3372
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3591..3695)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3763..3781
                     /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 cgagctcggt acccggggat
      421 cctaatacga ctcactatag gggaactcct actacaagtt gctaagaggc tattttttag
      481 ttcaaattac tcatacaatc atgttaaaat taatcgcagt gagcaacaaa atgctcacct
      541 gggtcgcagt aaccccagtt aacaaaacaa gggaggtaat tttgtgccca aaaaaagaat
      601 aaaagattta gctgaaattg ctcttgttgc agcaatttat ttcgcactca caattatatt
      661 ttcgtccatt tcgtttttac ccgttcaatt tcgaatcggg gaaattacga aatccattgt
      721 agtattcaat aaaaaatatg ctatttccat gatgatagga aatttttttg caaatttgtt
      781 tagcccattt gctggtgcaa tggaattaat ttttatgcct ctttcgaact taataggctg
      841 tacaattgga tactacattg gaagacttac tcacaaagcg ataggagcta tattcatagc
      901 cctttggatt gcagcatcag ttgcaattac tttaaaggtt tctgcaggca taccatttat
      961 tccgactttc ttaagcgtgg gagtagcgga aactgtactt ttggtaactg gatatttttt
     1021 gcttttcaca attgaaaaga aaggagttgt gaaatttgac agataaatat aaagagagaa
     1081 gatttgacat ttacggttac gaaaaaattg acaaagaagt tctagaatct attgaatatg
     1141 agtatcctga aaaaaatact atcgtggagt atattaccga tgaattttct tctgtttgcc
     1201 cttggacagg attacctgac aatgcaaaac ttactataag gtatataccc cacaaaaaac
     1261 ttgtagaact taaatcctta aaatattacc ttacatctta taggaatgta ggtatattgc
     1321 aagaacatgc aataaacaga attttagatg atttggtgga attcctgcag ccaaaattta
     1381 tggaaataat aggcgaattt caggaaagag gaggaatagc tacaagaatt atagcaaggt
     1441 atgaaaaaga ggagtattaa acttaaaagg ctgcctaaaa ttttgtaggc agctttttta
     1501 ttcattttag tttttcttca aaatgagttt acagagaacg atgaggaagc aagcttggcg
     1561 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     1621 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     1681 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     1741 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1801 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1861 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1921 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1981 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2041 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2101 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2161 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2221 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2281 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2341 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2401 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2461 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2521 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2581 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2641 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2701 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2761 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2821 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2881 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2941 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3001 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3061 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3121 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3181 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3241 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3301 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3361 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3421 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3481 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3541 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3601 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3661 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3721 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     3781 ccctttcgtc
//
