LOCUS       sequence-166627-        6583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle plasmid for sarA P1-mAmetrine expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6583)
  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 6583)
  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: pRN12
FEATURES             Location/Qualifiers
     source          1..6583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1019..1035)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1019..1035)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1032..1054)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1043..1059
                     /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(1067..1073,1074..1091,1092..1097))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1112..1133
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1250..1267)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1421..2009)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1501..1520)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2477..2856)
                     /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     2803..2822
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3041..3145)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3213..3231
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3269..3291)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3392..3411
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3605..3627
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3619..3636
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3620..3636
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6311..6325
                     /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 attaatggtt tcaaaaggtg
      301 aagaattatt tacaggtgtt gttccaattt tagttgaatt agatggtgat gttaatggtc
      361 ataaattttc agttcgtggt gaaggtgaag gtgatgcaac aaatggtaaa ttaacattaa
      421 aatttatttg tacatcaggt aaattaccag ttccatggcc aacattagtt acaacattat
      481 catatggtgt tcaatgtttt gcacgttatc cagatcatat gaaacaacat gattttttta
      541 aatcagcaat gccagaaggt tatgttcaag aacgtacaat ttcatttaaa gatgatggtt
      601 catatcgtac acgtgcagaa gttaaatttg aaggtgatac attagttaat cgtattgaat
      661 taaaaggtat tgattttaaa gaagatggta atattttagg tcataaatta gaatataata
      721 tgaatgtttg ggatgcatat attacagcag ataaacaaaa aaatggtatt aaagcaaatt
      781 ttaaaattga acataatgtt gaagatggtg gtgttcaatt agcagatgca tatcaacaaa
      841 atacaccaat tggtgatggt tcagttttat taccagataa tcattattta tcatttcaat
      901 caaaattatt taaagatcca aatgaacaac gtgatcatat ggttttatta gaatttgtta
      961 cagcagcagg tattacacca ggtatggatg aattatataa ataagaattc gtaatcatgt
     1021 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     1081 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     1141 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     1201 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     1261 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     1321 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     1381 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1441 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1501 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1561 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1621 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     1681 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1741 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1801 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1861 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1921 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1981 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2041 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2101 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2161 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2221 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     2281 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     2341 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     2401 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     2461 cagttaatag ttgcgcaacg ttgttgccat tgctacaggc atcgtgggtg tcacgctcgt
     2521 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     2581 ccatgttgtg caaaaaagcg gttagctctt tcggtcctcc gatcgttgtc agaagtaagt
     2641 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     2701 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     2761 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     2821 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     2881 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     2941 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3001 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     3061 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     3121 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     3181 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     3241 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     3301 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     3361 ttggcgggtg tcggggctgg cttaaactat gcggcatcag agcagattgt actgagagtg
     3421 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     3481 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     3541 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     3601 ttttcccagt cacgacgttg taaaacgacg gccagtgcca agcttctgta ggtttttagg
     3661 cataaaacta tatgatttac ccctaaatct ttaaaatgcc ccttaaaatt caaaataaag
     3721 gcatttaaaa tttaaatatt tcttgtgata aagtttgtta aaaaggagtg gttttatgac
     3781 tgttatgtgg ttatcgatta taggtatgtg gttttgtatt ggaatggcat tttttgctat
     3841 caaggttatt aaaaataaaa attagaccac gcatttatgc cgagaaaatt tattgtgcgt
     3901 tgagaagaac ccttaactaa acttgcagac gaatgtcggc atagcgtgag ctattaagcc
     3961 gaccattcga caagttttgg gattgttaag ggttccgagg ctcaacgtca ataaagcaat
     4021 tggaataaag aagcgaaaaa ggagaagtcg gttcagaaaa agaaggatat ggatctggag
     4081 ctgtaatata aaaaccttct tcaactaacg gggcaggtta gtgacattag aaaaccgact
     4141 gtaaaaagta cagtcggcat tatctcatat tataaaagcc agtcattagg cctatctgac
     4201 aattcctgaa tagagttcat aaacaatcct gcatgataac catcacaaac agaatgatgt
     4261 acctgtaaag atagcggtaa atatattgaa ttacctttat taatgaattt tcctgctgta
     4321 ataatgggta gaaggtaatt actattatta ttgatattta agttaaaccc agtaaatgaa
     4381 gtccatggaa taatagaaag agaaaaagca ttttcaggta taggtgtttt gggaaacaat
     4441 ttccccgaac cattatattt ctctacatca gaaaggtata aatcataaaa ctctttgaag
     4501 tcattcttta caggagtcca aataccagag aatgttttag atacaccatc aaaaattgta
     4561 taaagtggct ctaacttatc ccaataacct aactctccgt cgctattgta accagttcta
     4621 aaagctgtat ttgagtttat cacccttgtc actaagaaaa taaatgcagg gtaaaattta
     4681 tatccttctt gttttatgtt tcggtataaa acactaatat caatttctgt ggttatacta
     4741 aaagtcgttt gttggttcaa ataatgatta aatatctctt ttctcttcca attgtctaaa
     4801 tcaattttat taaagttcat ttgatatgcc tcctaaattt ttatctaaag tgaatttagg
     4861 aggcttactt gtctgctttc ttcattagaa tcaatccttt tttaaaagtc aatattactg
     4921 taacataaat atatatttta aaaatatccc actttatcca attttcgttt gttgaactaa
     4981 tgggtgcttt agttgaagaa taaaagacca cattaaaaaa tgtggtcttt tgtgtttttt
     5041 taaaggattt gagcgtagcg aaaaatcctt ttctttctta tcttgataat aagggtaact
     5101 attgccggcg aggctagtta cccttaagtt attggtatga ctggttttaa gcgcaaaaaa
     5161 agttgctttt tcgtacctat taatgtatcg ttttaaatga ctagtaaaaa acatacatag
     5221 aaaggggaaa aagcaacttt ttttattgtc atagtttgtg aaaactaagt tgtttttatg
     5281 tgttataaca tggaaaagta tactgagaaa aaacaaagaa atcaagtatt tcagaaattt
     5341 attaaacgtc atattggaga gaatcaaatg gatttagttg aagattgcaa tacatttctg
     5401 tcttttgtag ctgataaaac tttagaaaaa cagaaattat ataaagctaa ttcttgtaaa
     5461 aatcgatttt gtcctgtctg tgcttggaga aaagctagaa aagatgcatt gggtttatct
     5521 ttgatgatgc aatatattaa gcagcaagag aaaaaggagt ttatcttttt aactttgact
     5581 acacctaatg taatgagtga tgaattagaa aatgaaataa aacgttataa taattctttt
     5641 agaaaactta taaagagaaa aaaagtaggt agtgttataa agggatatgt tcgtaagtta
     5701 gagattacat ataataaaaa aagagatgat tataatcctc attttcatgt gttaattgca
     5761 gtaaataaat cgtatttcac agataaaaga tattatatta gccaacaaga atggttagat
     5821 ttatggcgtg atgtaacggg catttcagaa ataacacaag ttcaagttca aaaaataaga
     5881 caaaataata ataaagaatt atatgaaatg gctaagtatt ctggtaaaga tagtgattat
     5941 ttaataaatc aaaaagtctt tgatgcattt tataaatcac ttaaaggtaa acaggtatta
     6001 gtttattcag gattatttaa agaggctaaa aagaaattaa aaaatgggga tttagattac
     6061 ttaaaagaaa ttgatccaac cgaatatatc tatcaaattt tttatatttg gaaacaaaaa
     6121 gagtatttag ctagtgaact ttatgactta acagaacaag aaaaaagaga aattaatcac
     6181 aaaatgatag acgaaatcga ggaagaacaa taacaaaata taagtgctaa cagctgacct
     6241 cccgataaca ccatgtagtt attgggaggt cagctgttga attatgcacg agtattttaa
     6301 aagttattgt gatgacgacg ataaacgatt atcaaaagta taatgttaaa atgctttatt
     6361 atactaacgt tatataaaca ttatactttc gttatacaaa ttttaaccct gttaggaact
     6421 ataaaaaatc atgaaaattt taatttgcat gtaactgggc agtgtcttaa aaaatcgaca
     6481 ctgaatttgc tcaaattttt gtttgtagaa ttagaatata tttatttggc tcatatttgc
     6541 tttttaaaag cttgcatgcc tgcaggtcga ctctagagga tcc
//
