LOCUS       sequence-166624-        6574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle plasmid for sarA P1-mCherry expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6574)
  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 6574)
  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: pRN11
FEATURES             Location/Qualifiers
     source          1..6574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1010..1026)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1010..1026)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1023..1045)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1034..1050
                     /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(1058..1064,1065..1082,1083..1088))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1103..1124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1241..1258)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1412..2000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1492..1511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2468..2847)
                     /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     2794..2813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3032..3136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3204..3222
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3260..3282)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3383..3402
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3596..3618
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3610..3627
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3611..3627
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6302..6316
                     /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 gaggatgtat acatatggtg agcaagggcg
      301 aggaggataa tatggctatt atcaaagaat ttatgagatt caaagttcat atggaaggtt
      361 cagtcaatgg tcacgaattt gaaattgagg gagaaggtga aggcagacca tatgaaggta
      421 ctcaaacggc caaacttaaa gtcactaaag gaggtcctct tccatttgca tgggatattc
      481 ttagtccaca atttatgtat gggtctaaag cttacgttaa acatccagcg gacattccgg
      541 attatcttaa attgtcgttt ccagaaggct tcaaatggga aagagtaatg aactttgaag
      601 atggaggagt cgttacagtc actcaggata gttctcttca agatggagaa tttatctaca
      661 aagtcaagtt aagaggtacc aattttccct cagatggtcc agttatgcaa aagaaaacta
      721 tgggatggga agcatccagt gagcgtatgt atcctgaaga tggtgccctt aaaggcgaaa
      781 ttaaacaacg cctaaagctt aaagatggtg gacattatga cgctgaagtt aaaacaactt
      841 acaaagcgaa gaaaccggta caacttccag gtgcatataa tgtcaacatt aaacttgata
      901 ttacaagtca taatgaagat tatactattg ttgaacaata tgagagagct gaaggaagac
      961 attctacagg cggcatggac gagctgtaca agtaagaatt cgtaatcatg tcatagctgt
     1021 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     1081 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     1141 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     1201 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1261 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1321 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1381 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1441 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1501 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1561 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1621 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1681 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1741 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1801 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     1861 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1921 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1981 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2041 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2101 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2161 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2221 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2281 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     2341 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     2401 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2461 gttgcgcaac gttgttgcca ttgctacagg catcgtgggt gtcacgctcg tcgtttggta
     2521 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2581 gcaaaaaagc ggttagctct ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     2641 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     2701 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     2761 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     2821 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     2881 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     2941 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3001 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3061 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3121 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     3181 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     3241 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     3301 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     3361 gtcggggctg gcttaaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     3421 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     3481 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     3541 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     3601 tcacgacgtt gtaaaacgac ggccagtgcc aagcttctgt aggtttttag gcataaaact
     3661 atatgattta cccctaaatc tttaaaatgc cccttaaaat tcaaaataaa ggcatttaaa
     3721 atttaaatat ttcttgtgat aaagtttgtt aaaaaggagt ggttttatga ctgttatgtg
     3781 gttatcgatt ataggtatgt ggttttgtat tggaatggca ttttttgcta tcaaggttat
     3841 taaaaataaa aattagacca cgcatttatg ccgagaaaat ttattgtgcg ttgagaagaa
     3901 cccttaacta aacttgcaga cgaatgtcgg catagcgtga gctattaagc cgaccattcg
     3961 acaagttttg ggattgttaa gggttccgag gctcaacgtc aataaagcaa ttggaataaa
     4021 gaagcgaaaa aggagaagtc ggttcagaaa aagaaggata tggatctgga gctgtaatat
     4081 aaaaaccttc ttcaactaac ggggcaggtt agtgacatta gaaaaccgac tgtaaaaagt
     4141 acagtcggca ttatctcata ttataaaagc cagtcattag gcctatctga caattcctga
     4201 atagagttca taaacaatcc tgcatgataa ccatcacaaa cagaatgatg tacctgtaaa
     4261 gatagcggta aatatattga attaccttta ttaatgaatt ttcctgctgt aataatgggt
     4321 agaaggtaat tactattatt attgatattt aagttaaacc cagtaaatga agtccatgga
     4381 ataatagaaa gagaaaaagc attttcaggt ataggtgttt tgggaaacaa tttccccgaa
     4441 ccattatatt tctctacatc agaaaggtat aaatcataaa actctttgaa gtcattcttt
     4501 acaggagtcc aaataccaga gaatgtttta gatacaccat caaaaattgt ataaagtggc
     4561 tctaacttat cccaataacc taactctccg tcgctattgt aaccagttct aaaagctgta
     4621 tttgagttta tcacccttgt cactaagaaa ataaatgcag ggtaaaattt atatccttct
     4681 tgttttatgt ttcggtataa aacactaata tcaatttctg tggttatact aaaagtcgtt
     4741 tgttggttca aataatgatt aaatatctct tttctcttcc aattgtctaa atcaatttta
     4801 ttaaagttca tttgatatgc ctcctaaatt tttatctaaa gtgaatttag gaggcttact
     4861 tgtctgcttt cttcattaga atcaatcctt ttttaaaagt caatattact gtaacataaa
     4921 tatatatttt aaaaatatcc cactttatcc aattttcgtt tgttgaacta atgggtgctt
     4981 tagttgaaga ataaaagacc acattaaaaa atgtggtctt ttgtgttttt ttaaaggatt
     5041 tgagcgtagc gaaaaatcct tttctttctt atcttgataa taagggtaac tattgccggc
     5101 gaggctagtt acccttaagt tattggtatg actggtttta agcgcaaaaa aagttgcttt
     5161 ttcgtaccta ttaatgtatc gttttaaatg actagtaaaa aacatacata gaaaggggaa
     5221 aaagcaactt tttttattgt catagtttgt gaaaactaag ttgtttttat gtgttataac
     5281 atggaaaagt atactgagaa aaaacaaaga aatcaagtat ttcagaaatt tattaaacgt
     5341 catattggag agaatcaaat ggatttagtt gaagattgca atacatttct gtcttttgta
     5401 gctgataaaa ctttagaaaa acagaaatta tataaagcta attcttgtaa aaatcgattt
     5461 tgtcctgtct gtgcttggag aaaagctaga aaagatgcat tgggtttatc tttgatgatg
     5521 caatatatta agcagcaaga gaaaaaggag tttatctttt taactttgac tacacctaat
     5581 gtaatgagtg atgaattaga aaatgaaata aaacgttata ataattcttt tagaaaactt
     5641 ataaagagaa aaaaagtagg tagtgttata aagggatatg ttcgtaagtt agagattaca
     5701 tataataaaa aaagagatga ttataatcct cattttcatg tgttaattgc agtaaataaa
     5761 tcgtatttca cagataaaag atattatatt agccaacaag aatggttaga tttatggcgt
     5821 gatgtaacgg gcatttcaga aataacacaa gttcaagttc aaaaaataag acaaaataat
     5881 aataaagaat tatatgaaat ggctaagtat tctggtaaag atagtgatta tttaataaat
     5941 caaaaagtct ttgatgcatt ttataaatca cttaaaggta aacaggtatt agtttattca
     6001 ggattattta aagaggctaa aaagaaatta aaaaatgggg atttagatta cttaaaagaa
     6061 attgatccaa ccgaatatat ctatcaaatt ttttatattt ggaaacaaaa agagtattta
     6121 gctagtgaac tttatgactt aacagaacaa gaaaaaagag aaattaatca caaaatgata
     6181 gacgaaatcg aggaagaaca ataacaaaat ataagtgcta acagctgacc tcccgataac
     6241 accatgtagt tattgggagg tcagctgttg aattatgcac gagtatttta aaagttattg
     6301 tgatgacgac gataaacgat tatcaaaagt ataatgttaa aatgctttat tatactaacg
     6361 ttatataaac attatacttt cgttatacaa attttaaccc tgttaggaac tataaaaaat
     6421 catgaaaatt ttaatttgca tgtaactggg cagtgtctta aaaaatcgac actgaatttg
     6481 ctcaaatttt tgtttgtaga attagaatat atttatttgg ctcatatttg ctttttaaaa
     6541 gcttgcatgc ctgcaggtcg actctagagg atcc
//
