LOCUS       sequence-163494-        6559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle plasmid for sarA P1-mKate expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6559)
  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 6559)
  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: pRN10
FEATURES             Location/Qualifiers
     source          1..6559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(730..746)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(730..746)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(743..765)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    754..770
                     /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(778..784,785..802,803..808))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    823..844
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(961..978)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1132..1720)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1212..1231)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2188..2567)
                     /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     2514..2533
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2752..2856)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2924..2942
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2980..3002)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3103..3122
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3316..3338
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3330..3347
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3331..3347
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6022..6036
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
ORIGIN
        1 cttaggagga tgattattta tgtcagaact tatcaaggaa aatatgcaca tgaaattgta
       61 catggaagga acagtaaata atcaccactt taaatgtacc tcagaaggag aaggaaaacc
      121 atatgaaggt actcaaacca tgcgtattaa ggccgttgaa ggtggaccat tgccttttgc
      181 ctttgatatt cttgctacat cttttatgta cggatcaaag acttttatca atcataccca
      241 aggtatccca gatttcttta aacagtcatt tcctgaagga tttacatggg aacgtgtcac
      301 aacttatgaa gatggtggag tattgacagc aactcaagat acatctttgc aagatggttg
      361 tcttatctac aatgtaaaga tccgtggagt taattttcca agtaatggtc ctgttatgca
      421 gaaaaagacc cttggatggg aagcatctac cgaaacatta tatcctgccg atggtggatt
      481 ggaaggtcgt gctgatatgg cattgaaact tgtcggtgga ggtcacctta tctgtaattt
      541 gaagaccaca taccgttcta aaaagccagc taagaatctt aagatgcctg gtgtttacta
      601 cgtcgatcgt cgtttagaac gtatcaaaga agcagataag gaaacttatg ttgaacagca
      661 cgaagtagcc gtcgcacgtt attgtgattt gcctagtaaa ttgggacacc gttaagaatt
      721 cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
      781 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
      841 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
      901 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
      961 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1021 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1081 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1141 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1201 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1261 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1321 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1381 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1441 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1501 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1561 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1621 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1681 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1741 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1801 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1861 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     1921 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     1981 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2041 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2101 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2161 aagtagttcg ccagttaata gttgcgcaac gttgttgcca ttgctacagg catcgtgggt
     2221 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2281 tacatgatcc cccatgttgt gcaaaaaagc ggttagctct ttcggtcctc cgatcgttgt
     2341 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2401 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2461 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     2521 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2581 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2641 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2701 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     2761 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     2821 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     2881 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     2941 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     3001 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     3061 gtcagcgggt gttggcgggt gtcggggctg gcttaaacta tgcggcatca gagcagattg
     3121 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     3181 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     3241 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     3301 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagcttctgt
     3361 aggtttttag gcataaaact atatgattta cccctaaatc tttaaaatgc cccttaaaat
     3421 tcaaaataaa ggcatttaaa atttaaatat ttcttgtgat aaagtttgtt aaaaaggagt
     3481 ggttttatga ctgttatgtg gttatcgatt ataggtatgt ggttttgtat tggaatggca
     3541 ttttttgcta tcaaggttat taaaaataaa aattagacca cgcatttatg ccgagaaaat
     3601 ttattgtgcg ttgagaagaa cccttaacta aacttgcaga cgaatgtcgg catagcgtga
     3661 gctattaagc cgaccattcg acaagttttg ggattgttaa gggttccgag gctcaacgtc
     3721 aataaagcaa ttggaataaa gaagcgaaaa aggagaagtc ggttcagaaa aagaaggata
     3781 tggatctgga gctgtaatat aaaaaccttc ttcaactaac ggggcaggtt agtgacatta
     3841 gaaaaccgac tgtaaaaagt acagtcggca ttatctcata ttataaaagc cagtcattag
     3901 gcctatctga caattcctga atagagttca taaacaatcc tgcatgataa ccatcacaaa
     3961 cagaatgatg tacctgtaaa gatagcggta aatatattga attaccttta ttaatgaatt
     4021 ttcctgctgt aataatgggt agaaggtaat tactattatt attgatattt aagttaaacc
     4081 cagtaaatga agtccatgga ataatagaaa gagaaaaagc attttcaggt ataggtgttt
     4141 tgggaaacaa tttccccgaa ccattatatt tctctacatc agaaaggtat aaatcataaa
     4201 actctttgaa gtcattcttt acaggagtcc aaataccaga gaatgtttta gatacaccat
     4261 caaaaattgt ataaagtggc tctaacttat cccaataacc taactctccg tcgctattgt
     4321 aaccagttct aaaagctgta tttgagttta tcacccttgt cactaagaaa ataaatgcag
     4381 ggtaaaattt atatccttct tgttttatgt ttcggtataa aacactaata tcaatttctg
     4441 tggttatact aaaagtcgtt tgttggttca aataatgatt aaatatctct tttctcttcc
     4501 aattgtctaa atcaatttta ttaaagttca tttgatatgc ctcctaaatt tttatctaaa
     4561 gtgaatttag gaggcttact tgtctgcttt cttcattaga atcaatcctt ttttaaaagt
     4621 caatattact gtaacataaa tatatatttt aaaaatatcc cactttatcc aattttcgtt
     4681 tgttgaacta atgggtgctt tagttgaaga ataaaagacc acattaaaaa atgtggtctt
     4741 ttgtgttttt ttaaaggatt tgagcgtagc gaaaaatcct tttctttctt atcttgataa
     4801 taagggtaac tattgccggc gaggctagtt acccttaagt tattggtatg actggtttta
     4861 agcgcaaaaa aagttgcttt ttcgtaccta ttaatgtatc gttttaaatg actagtaaaa
     4921 aacatacata gaaaggggaa aaagcaactt tttttattgt catagtttgt gaaaactaag
     4981 ttgtttttat gtgttataac atggaaaagt atactgagaa aaaacaaaga aatcaagtat
     5041 ttcagaaatt tattaaacgt catattggag agaatcaaat ggatttagtt gaagattgca
     5101 atacatttct gtcttttgta gctgataaaa ctttagaaaa acagaaatta tataaagcta
     5161 attcttgtaa aaatcgattt tgtcctgtct gtgcttggag aaaagctaga aaagatgcat
     5221 tgggtttatc tttgatgatg caatatatta agcagcaaga gaaaaaggag tttatctttt
     5281 taactttgac tacacctaat gtaatgagtg atgaattaga aaatgaaata aaacgttata
     5341 ataattcttt tagaaaactt ataaagagaa aaaaagtagg tagtgttata aagggatatg
     5401 ttcgtaagtt agagattaca tataataaaa aaagagatga ttataatcct cattttcatg
     5461 tgttaattgc agtaaataaa tcgtatttca cagataaaag atattatatt agccaacaag
     5521 aatggttaga tttatggcgt gatgtaacgg gcatttcaga aataacacaa gttcaagttc
     5581 aaaaaataag acaaaataat aataaagaat tatatgaaat ggctaagtat tctggtaaag
     5641 atagtgatta tttaataaat caaaaagtct ttgatgcatt ttataaatca cttaaaggta
     5701 aacaggtatt agtttattca ggattattta aagaggctaa aaagaaatta aaaaatgggg
     5761 atttagatta cttaaaagaa attgatccaa ccgaatatat ctatcaaatt ttttatattt
     5821 ggaaacaaaa agagtattta gctagtgaac tttatgactt aacagaacaa gaaaaaagag
     5881 aaattaatca caaaatgata gacgaaatcg aggaagaaca ataacaaaat ataagtgcta
     5941 acagctgacc tcccgataac accatgtagt tattgggagg tcagctgttg aattatgcac
     6001 gagtatttta aaagttattg tgatgacgac gataaacgat tatcaaaagt ataatgttaa
     6061 aatgctttat tatactaacg ttatataaac attatacttt cgttatacaa attttaaccc
     6121 tgttaggaac tataaaaaat catgaaaatt ttaatttgca tgtaactggg cagtgtctta
     6181 aaaaatcgac actgaatttg ctcaaatttt tgtttgtaga attagaatat atttatttgg
     6241 ctcatatttg ctttttaaaa gcttgcatgc ctgcaggtcg actctagagg atccgctagc
     6301 ctgatatttt tgactaaacc aaatgctaac ccagaaatac aatcactgtg tctaatgaat
     6361 aatttgtttt ataaacactt ttttgtttac ttctcatttt taattagtta taattaacta
     6421 aataatagag cattaaatat atttaataaa acttatttaa tgcaaaatta tgactaacat
     6481 atctataata aataaagatt agatatcaat atattatcgg gcaaatgtat cgagcaagat
     6541 gcatcggatc cccgggtac
//
