LOCUS       sequence-163430-        6537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle plasmid for sarA P1-dsRED expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6537)
  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 6537)
  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: pTH1
FEATURES             Location/Qualifiers
     source          1..6537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             281..958
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MDNTEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEPSTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(411..429)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(461..481)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     679..698
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     873..896
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(973..989)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(973..989)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(986..1008)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    997..1013
                     /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(1021..1027,1028..1045,1046..1051))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1066..1087
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1204..1221)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1375..1963)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1455..1474)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2431..2810)
                     /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     2757..2776
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2995..3099)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3167..3185
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3223..3245)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3346..3365
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3559..3581
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3573..3590
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3574..3590
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6265..6279
                     /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 ggtaccagga ggaaaaacat atggacaaca ccgaggacgt
      301 catcaaggag ttcatgcgct tcaaggtgcg catggagggc tccgtgaacg gccacgagtt
      361 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
      421 ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttccagta
      481 cggctccaag gtgtacgtga agcaccccgc cgacatcccc gactacaaga agctgtcctt
      541 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
      601 gacccaggac tcctccctgc aggacggctg cttcatctac aaggtgaagt tcatcggcgt
      661 gaacttcccc tccgacggcc ccgtaatgca gaagaagact atgggctggg agccctccac
      721 cgagcgcctg tacccccgcg acggcgtgct gaagggcgag atccacaagg ccctgaagct
      781 gaaggacggc ggccactacc tggtggagtt caagtccatc tacatggcca agaagcccgt
      841 gcagctgccc ggctactact acgtggactc caagctggac atcacctccc acaacgagga
      901 ctacaccatc gtggagcagt acgagcgcac cgagggccgc caccacctgt tcctgtagga
      961 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     1021 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     1081 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     1141 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1201 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1261 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1321 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1381 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1441 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1501 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1561 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1621 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1681 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1741 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1801 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1861 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1921 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1981 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2041 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2101 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2161 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2221 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2281 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2341 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2401 agtaagtagt tcgccagtta atagttgcgc aacgttgttg ccattgctac aggcatcgtg
     2461 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2521 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tctttcggtc ctccgatcgt
     2581 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2641 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     2701 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     2761 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     2821 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     2881 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     2941 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3001 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3061 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3121 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     3181 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     3241 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     3301 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaaa ctatgcggca tcagagcaga
     3361 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     3421 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     3481 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     3541 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagcttc
     3601 tgtaggtttt taggcataaa actatatgat ttacccctaa atctttaaaa tgccccttaa
     3661 aattcaaaat aaaggcattt aaaatttaaa tatttcttgt gataaagttt gttaaaaagg
     3721 agtggtttta tgactgttat gtggttatcg attataggta tgtggttttg tattggaatg
     3781 gcattttttg ctatcaaggt tattaaaaat aaaaattaga ccacgcattt atgccgagaa
     3841 aatttattgt gcgttgagaa gaacccttaa ctaaacttgc agacgaatgt cggcatagcg
     3901 tgagctatta agccgaccat tcgacaagtt ttgggattgt taagggttcc gaggctcaac
     3961 gtcaataaag caattggaat aaagaagcga aaaaggagaa gtcggttcag aaaaagaagg
     4021 atatggatct ggagctgtaa tataaaaacc ttcttcaact aacggggcag gttagtgaca
     4081 ttagaaaacc gactgtaaaa agtacagtcg gcattatctc atattataaa agccagtcat
     4141 taggcctatc tgacaattcc tgaatagagt tcataaacaa tcctgcatga taaccatcac
     4201 aaacagaatg atgtacctgt aaagatagcg gtaaatatat tgaattacct ttattaatga
     4261 attttcctgc tgtaataatg ggtagaaggt aattactatt attattgata tttaagttaa
     4321 acccagtaaa tgaagtccat ggaataatag aaagagaaaa agcattttca ggtataggtg
     4381 ttttgggaaa caatttcccc gaaccattat atttctctac atcagaaagg tataaatcat
     4441 aaaactcttt gaagtcattc tttacaggag tccaaatacc agagaatgtt ttagatacac
     4501 catcaaaaat tgtataaagt ggctctaact tatcccaata acctaactct ccgtcgctat
     4561 tgtaaccagt tctaaaagct gtatttgagt ttatcaccct tgtcactaag aaaataaatg
     4621 cagggtaaaa tttatatcct tcttgtttta tgtttcggta taaaacacta atatcaattt
     4681 ctgtggttat actaaaagtc gtttgttggt tcaaataatg attaaatatc tcttttctct
     4741 tccaattgtc taaatcaatt ttattaaagt tcatttgata tgcctcctaa atttttatct
     4801 aaagtgaatt taggaggctt acttgtctgc tttcttcatt agaatcaatc cttttttaaa
     4861 agtcaatatt actgtaacat aaatatatat tttaaaaata tcccacttta tccaattttc
     4921 gtttgttgaa ctaatgggtg ctttagttga agaataaaag accacattaa aaaatgtggt
     4981 cttttgtgtt tttttaaagg atttgagcgt agcgaaaaat ccttttcttt cttatcttga
     5041 taataagggt aactattgcc ggcgaggcta gttaccctta agttattggt atgactggtt
     5101 ttaagcgcaa aaaaagttgc tttttcgtac ctattaatgt atcgttttaa atgactagta
     5161 aaaaacatac atagaaaggg gaaaaagcaa ctttttttat tgtcatagtt tgtgaaaact
     5221 aagttgtttt tatgtgttat aacatggaaa agtatactga gaaaaaacaa agaaatcaag
     5281 tatttcagaa atttattaaa cgtcatattg gagagaatca aatggattta gttgaagatt
     5341 gcaatacatt tctgtctttt gtagctgata aaactttaga aaaacagaaa ttatataaag
     5401 ctaattcttg taaaaatcga ttttgtcctg tctgtgcttg gagaaaagct agaaaagatg
     5461 cattgggttt atctttgatg atgcaatata ttaagcagca agagaaaaag gagtttatct
     5521 ttttaacttt gactacacct aatgtaatga gtgatgaatt agaaaatgaa ataaaacgtt
     5581 ataataattc ttttagaaaa cttataaaga gaaaaaaagt aggtagtgtt ataaagggat
     5641 atgttcgtaa gttagagatt acatataata aaaaaagaga tgattataat cctcattttc
     5701 atgtgttaat tgcagtaaat aaatcgtatt tcacagataa aagatattat attagccaac
     5761 aagaatggtt agatttatgg cgtgatgtaa cgggcatttc agaaataaca caagttcaag
     5821 ttcaaaaaat aagacaaaat aataataaag aattatatga aatggctaag tattctggta
     5881 aagatagtga ttatttaata aatcaaaaag tctttgatgc attttataaa tcacttaaag
     5941 gtaaacaggt attagtttat tcaggattat ttaaagaggc taaaaagaaa ttaaaaaatg
     6001 gggatttaga ttacttaaaa gaaattgatc caaccgaata tatctatcaa attttttata
     6061 tttggaaaca aaaagagtat ttagctagtg aactttatga cttaacagaa caagaaaaaa
     6121 gagaaattaa tcacaaaatg atagacgaaa tcgaggaaga acaataacaa aatataagtg
     6181 ctaacagctg acctcccgat aacaccatgt agttattggg aggtcagctg ttgaattatg
     6241 cacgagtatt ttaaaagtta ttgtgatgac gacgataaac gattatcaaa agtataatgt
     6301 taaaatgctt tattatacta acgttatata aacattatac tttcgttata caaattttaa
     6361 ccctgttagg aactataaaa aatcatgaaa attttaattt gcatgtaact gggcagtgtc
     6421 ttaaaaaatc gacactgaat ttgctcaaat ttttgtttgt agaattagaa tatatttatt
     6481 tggctcatat ttgcttttta aaagcttgca tgcctgcagg tcgactctag aggatcc
//
