LOCUS       sequence-163677-        6580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle plasmid for sarA P1-YFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6580)
  AUTHORS   de Jong NW, van der Horst T, van Strijp JA, Nijland R
  TITLE     Fluorescent reporters for markerless genomic integration in 
            Staphylococcus aureus.
  JOURNAL   Sci Rep. 2017 Mar 7;7:43889. doi: 10.1038/srep43889.
  PUBMED    28266573
REFERENCE   2  (bases 1 to 6580)
  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: "Sci Rep."; 
            date: "2017-03-7"; volume: "7:43889. doi"; pages: " 
            10.1038/srep43889"
COMMENT     Sequence Label: pTH3
FEATURES             Location/Qualifiers
     source          1..6580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(315..343)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     938..958
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1016..1032)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1016..1032)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1029..1051)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1040..1056
                     /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(1064..1070,1071..1088,1089..1094))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1109..1130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1247..1264)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1418..2006)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1498..1517)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2474..2853)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     2800..2819
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3038..3142)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3210..3228
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3266..3288)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3389..3408
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3602..3624
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3616..3633
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3617..3633
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6308..6322
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
ORIGIN
        1 gctagcctga tatttttgac taaaccaaat gctaacccag aaatacaatc actgtgtcta
       61 atgaataatt tgttttataa acactttttt gtttacttct catttttaat tagttataat
      121 taactaaata atagagcatt aaatatattt aataaaactt atttaatgca aaattatgac
      181 taacatatct ataataaata aagattagat atcaatatat tatcgggcaa atgtatcgag
      241 caagatgcat cggatccccg ggtaccttag gaggatgatt atttatgagt aaaggagaag
      301 aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca
      361 aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaac
      421 ttatttgcac tactggaaaa ctacctgttc catggccaac acttgtcact actcttggat
      481 atggtcttca atgctttgcg agatacccag atcatatgaa acagcatgac tttttcaaga
      541 gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact
      601 acaagacacg tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa
      661 aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata
      721 actcacacaa tgtatacatc acagcagaca aacaaaagaa tggaatcaaa gctaacttca
      781 aaattagaca caacattgaa gatggaggtg ttcaactagc agaccattat caacaaaata
      841 ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc tatcaatctg
      901 ccctttcgaa agatcccaac gaaaagagag accacatggt ccttcttgag tttgtaacag
      961 ctgctgggat tacacttggc atggacgagc tgtacaagta agaattcgta atcatgtcat
     1021 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     1081 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     1141 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     1201 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     1261 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     1321 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     1381 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1441 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1501 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1561 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1621 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1681 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1741 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1801 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     1861 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1921 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1981 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2041 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2101 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     2161 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     2221 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     2281 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     2341 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     2401 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     2461 ttaatagttg cgcaacgttg ttgccattgc tacaggcatc gtgggtgtca cgctcgtcgt
     2521 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     2581 tgttgtgcaa aaaagcggtt agctctttcg gtcctccgat cgttgtcaga agtaagttgg
     2641 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     2701 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     2761 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     2821 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     2881 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     2941 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3001 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     3061 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3121 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     3181 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     3241 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     3301 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     3361 gcgggtgtcg gggctggctt aaactatgcg gcatcagagc agattgtact gagagtgcac
     3421 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     3481 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     3541 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     3601 tcccagtcac gacgttgtaa aacgacggcc agtgccaagc ttctgtaggt ttttaggcat
     3661 aaaactatat gatttacccc taaatcttta aaatgcccct taaaattcaa aataaaggca
     3721 tttaaaattt aaatatttct tgtgataaag tttgttaaaa aggagtggtt ttatgactgt
     3781 tatgtggtta tcgattatag gtatgtggtt ttgtattgga atggcatttt ttgctatcaa
     3841 ggttattaaa aataaaaatt agaccacgca tttatgccga gaaaatttat tgtgcgttga
     3901 gaagaaccct taactaaact tgcagacgaa tgtcggcata gcgtgagcta ttaagccgac
     3961 cattcgacaa gttttgggat tgttaagggt tccgaggctc aacgtcaata aagcaattgg
     4021 aataaagaag cgaaaaagga gaagtcggtt cagaaaaaga aggatatgga tctggagctg
     4081 taatataaaa accttcttca actaacgggg caggttagtg acattagaaa accgactgta
     4141 aaaagtacag tcggcattat ctcatattat aaaagccagt cattaggcct atctgacaat
     4201 tcctgaatag agttcataaa caatcctgca tgataaccat cacaaacaga atgatgtacc
     4261 tgtaaagata gcggtaaata tattgaatta cctttattaa tgaattttcc tgctgtaata
     4321 atgggtagaa ggtaattact attattattg atatttaagt taaacccagt aaatgaagtc
     4381 catggaataa tagaaagaga aaaagcattt tcaggtatag gtgttttggg aaacaatttc
     4441 cccgaaccat tatatttctc tacatcagaa aggtataaat cataaaactc tttgaagtca
     4501 ttctttacag gagtccaaat accagagaat gttttagata caccatcaaa aattgtataa
     4561 agtggctcta acttatccca ataacctaac tctccgtcgc tattgtaacc agttctaaaa
     4621 gctgtatttg agtttatcac ccttgtcact aagaaaataa atgcagggta aaatttatat
     4681 ccttcttgtt ttatgtttcg gtataaaaca ctaatatcaa tttctgtggt tatactaaaa
     4741 gtcgtttgtt ggttcaaata atgattaaat atctcttttc tcttccaatt gtctaaatca
     4801 attttattaa agttcatttg atatgcctcc taaattttta tctaaagtga atttaggagg
     4861 cttacttgtc tgctttcttc attagaatca atcctttttt aaaagtcaat attactgtaa
     4921 cataaatata tattttaaaa atatcccact ttatccaatt ttcgtttgtt gaactaatgg
     4981 gtgctttagt tgaagaataa aagaccacat taaaaaatgt ggtcttttgt gtttttttaa
     5041 aggatttgag cgtagcgaaa aatccttttc tttcttatct tgataataag ggtaactatt
     5101 gccggcgagg ctagttaccc ttaagttatt ggtatgactg gttttaagcg caaaaaaagt
     5161 tgctttttcg tacctattaa tgtatcgttt taaatgacta gtaaaaaaca tacatagaaa
     5221 ggggaaaaag caactttttt tattgtcata gtttgtgaaa actaagttgt ttttatgtgt
     5281 tataacatgg aaaagtatac tgagaaaaaa caaagaaatc aagtatttca gaaatttatt
     5341 aaacgtcata ttggagagaa tcaaatggat ttagttgaag attgcaatac atttctgtct
     5401 tttgtagctg ataaaacttt agaaaaacag aaattatata aagctaattc ttgtaaaaat
     5461 cgattttgtc ctgtctgtgc ttggagaaaa gctagaaaag atgcattggg tttatctttg
     5521 atgatgcaat atattaagca gcaagagaaa aaggagttta tctttttaac tttgactaca
     5581 cctaatgtaa tgagtgatga attagaaaat gaaataaaac gttataataa ttcttttaga
     5641 aaacttataa agagaaaaaa agtaggtagt gttataaagg gatatgttcg taagttagag
     5701 attacatata ataaaaaaag agatgattat aatcctcatt ttcatgtgtt aattgcagta
     5761 aataaatcgt atttcacaga taaaagatat tatattagcc aacaagaatg gttagattta
     5821 tggcgtgatg taacgggcat ttcagaaata acacaagttc aagttcaaaa aataagacaa
     5881 aataataata aagaattata tgaaatggct aagtattctg gtaaagatag tgattattta
     5941 ataaatcaaa aagtctttga tgcattttat aaatcactta aaggtaaaca ggtattagtt
     6001 tattcaggat tatttaaaga ggctaaaaag aaattaaaaa atggggattt agattactta
     6061 aaagaaattg atccaaccga atatatctat caaatttttt atatttggaa acaaaaagag
     6121 tatttagcta gtgaacttta tgacttaaca gaacaagaaa aaagagaaat taatcacaaa
     6181 atgatagacg aaatcgagga agaacaataa caaaatataa gtgctaacag ctgacctccc
     6241 gataacacca tgtagttatt gggaggtcag ctgttgaatt atgcacgagt attttaaaag
     6301 ttattgtgat gacgacgata aacgattatc aaaagtataa tgttaaaatg ctttattata
     6361 ctaacgttat ataaacatta tactttcgtt atacaaattt taaccctgtt aggaactata
     6421 aaaaatcatg aaaattttaa tttgcatgta actgggcagt gtcttaaaaa atcgacactg
     6481 aatttgctca aatttttgtt tgtagaatta gaatatattt atttggctca tatttgcttt
     6541 ttaaaagctt gcatgcctgc aggtcgactc tagaggatcc
//
