LOCUS       sequence-98510-e        3716 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7-transcription template for Tte mRNA used for in vitro 
            translation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3716)
  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 3716)
  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_TTE1564
FEATURES             Location/Qualifiers
     source          1..3716
                     /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(1495..1511)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1495..1511)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1508..1530)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1519..1535
                     /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(1543..1549,1550..1567,1568..1573))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1588..1609
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1726..1743)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1897..2485)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1977..1996)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2656..3447,3448..3516))
                     /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     3279..3298
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3517..3621)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3689..3707
                     /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 ggcagtgagc aacaaaatgc tcacctgggt cgcagtaacc
      481 ccagttaaca aaacaaggga ggtaattttg tgcccaaaaa aagaataaaa gatttagctg
      541 aaattgctct tgttgcagca atttatttcg cactcacaat tatattttcg tccatttcgt
      601 ttttacccgt tcaatttcga atcggggaaa ttacgaaatc cattgtagta ttcaataaaa
      661 aatatgctat ttccatgatg ataggaaatt tttttgcaaa tttgtttagc ccatttgctg
      721 gtgcaatgga attaattttt atgcctcttt cgaacttaat aggctgtaca attggatact
      781 acattggaag acttactcac aaagcgatag gagctatatt catagccctt tggattgcag
      841 catcagttgc aattacttta aaggtttctg caggcatacc atttattccg actttcttaa
      901 gcgtgggagt agcggaaact gtacttttgg taactggata ttttttgctt ttcacaattg
      961 aaaagaaagg agttgtgaaa tttgacagat aaatataaag agagaagatt tgacatttac
     1021 ggttacgaaa aaattgacaa agaagttcta gaatctattg aatatgagta tcctgaaaaa
     1081 aatactatcg tggagtatat taccgatgaa ttttcttctg tttgcccttg gacaggatta
     1141 cctgacaatg caaaacttac tataaggtat ataccccaca aaaaacttgt agaacttaaa
     1201 tccttaaaat attaccttac atcttatagg aatgtaggta tattgcaaga acatgcaata
     1261 aacagaattt tagatgattt ggtggaattc ctgcagccaa aatttatgga aataataggc
     1321 gaatttcagg aaagaggagg aatagctaca agaattatag caaggtatga aaaagaggag
     1381 tattaaactt aaaaggctgc ctaaaatttt gtaggcagct tttttattca ttttagtttt
     1441 tcttcaaaat gagtttacag agaacgatga ggaagcaagc ttggcgtaat catggtcata
     1501 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     1561 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1621 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1681 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1741 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1801 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1861 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1921 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1981 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2041 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2101 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2161 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2221 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2281 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2341 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2401 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2461 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2521 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2581 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2641 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2701 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2761 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2821 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     2881 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2941 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3001 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3061 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3121 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3181 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3241 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3301 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3361 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3421 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3481 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3541 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3601 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     3661 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
