LOCUS       sequence-163710-        9948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pJB38-NWMN29-30+ SarA_P1-mAmetrine-Term.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9948)
  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 9948)
  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: pRN112
FEATURES             Location/Qualifiers
     source          1..9948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2058..2074)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2058..2074)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      2083..2117
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     protein_bind    4944..4962
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    5033..5051
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             5073..5699
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(5149..5168)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(7690..7712)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    7798..7938
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7853..7872
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7953..7970)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8124..8712)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8204..8223)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8883..9674,9675..9743))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     9506..9525
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9744..9848)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9916..9934
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gaattcagtg gctacattcg aacatatcaa agcatacgat agaaaaacaa taagttatga
       61 tatgttacat gtaacaccac ctatgggtcc cttagatgta gtaaaagaaa gtacactttc
      121 agatagtgag ggttgggtag atgttaaccc aaccacatta cagcataaaa gctactctaa
      181 tgtatttgca cttggtgatg cttcaaatgt acctacttca aaaacaggcg cagctattcg
      241 taagcaagca cctatcgtcg ctaataattt attgcaagtg atgaataatc aaatgttaac
      301 gcatcattat gatggttata cttcatgccc tattgttact ggatataata ggttaatact
      361 tgcagagttt gattataata aaaatactaa agaaacaatg ccgtttaatc aggccaaaga
      421 acgtagaagt atgtatatat ttaagaaaga tttattacct aaaatgtatt ggtacggcat
      481 gctaaaagga ttaatataat aaagtacaga aaacaataaa tttttaatga aaaatctttt
      541 actataaaag attaagtatt taaatgacgt gtcagtgttg tgtttatatg tcgtgaattt
      601 ttagctctaa atagtataag attgaaaaag ttgttactgt tttaaatgat cacgatgaag
      661 tcattcaata agaatgatta tgaaaataga aacagcagta agatattttc taattgaaaa
      721 tcatctcact gctgtttttt aaaggtttat acctcatcct ctaaattatt taaaaataat
      781 taatggtatt tgagcacgtt tagcgacttt atgactgaca ttaccaattt ccatttcttg
      841 ccagatattc aaaccacgtg tactcaaaat gatagcttgg tatgtacctc caatagtaat
      901 ttcaataact ttgtctgttg aacactaaga gcaattttaa tttcataatg tgttgtaaac
      961 attttttttg attggagttt ttttctgagt taaacgatat atcgcgattg catgcctgca
     1021 ggtcgactct agaggatccg ctagcctgat atttttgact aaaccaaatg ctaacccaga
     1081 aatacaatca ctgtgtctaa tgaataattt gttttataaa cacttttttg tttacttctc
     1141 atttttaatt agttataatt aactaaataa tagagcatta aatatattta ataaaactta
     1201 tttaatgcaa aattatgact aacatatcta taataaataa agattagata tcaatatatt
     1261 atcgggcaaa tgtatcgagc aagatgcatc ggatccccgg gtaccttagg aggatgatta
     1321 tttatggttt caaaaggtga agaattattt acaggtgttg ttccaatttt agttgaatta
     1381 gatggtgatg ttaatggtca taaattttca gttcgtggtg aaggtgaagg tgatgcaaca
     1441 aatggtaaat taacattaaa atttatttgt acatcaggta aattaccagt tccatggcca
     1501 acattagtta caacattatc atatggtgtt caatgttttg cacgttatcc agatcatatg
     1561 aaacaacatg atttttttaa atcagcaatg ccagaaggtt atgttcaaga acgtacaatt
     1621 tcatttaaag atgatggttc atatcgtaca cgtgcagaag ttaaatttga aggtgataca
     1681 ttagttaatc gtattgaatt aaaaggtatt gattttaaag aagatggtaa tattttaggt
     1741 cataaattag aatataatat gaatgtttgg gatgcatata ttacagcaga taaacaaaaa
     1801 aatggtatta aagcaaattt taaaattgaa cataatgttg aagatggtgg tgttcaatta
     1861 gcagatgcat atcaacaaaa tacaccaatt ggtgatggtt cagttttatt accagataat
     1921 cattatttat catttcaatc aaaattattt aaagatccaa atgaacaacg tgatcatatg
     1981 gttttattag aatttgttac agcagcaggt attacaccag gtatggatga attatataaa
     2041 taagaattcg taatcatgtc atagctgttt cctgtgtgtt aattcagaac gctcggttgc
     2101 cgccgggcgt tttttatgca gctcgcgata tatcctgatg tatttttaat tttgcaccat
     2161 ttccaaaagg ataagtgaca taagtaaaaa ggcatcatcg ggagttatcc tatcaggaaa
     2221 accaagataa tacctaagta gaaagtgttc aatccgtgtt aaattgggaa atatcatcca
     2281 taaactttat tactcatact ataattcaat tttaacgtct tcgtccattt gggcttcaaa
     2341 ttcatcgagt agtgctcgtg cttctgcaat tgattgtgtg ttcatcaatt gatgtcgaag
     2401 ttcgctagcg cctcttatgc cacgcacata gattttaaag aatctacgca agctcttgaa
     2461 ttgtcgtatt tcatcttttt catatttgtt aaacaatgat aaatgcaatc tcaatagatc
     2521 taatagttcc ttgcttgtgt gttcgcgtgg ttctttttca aaagcgaatg gattgtggaa
     2581 aatgcctcta ccaatcatga cgccatcaat gccatatttt tctgccagtt caagtcctgt
     2641 ttttctatcg ggaatatcac cgttaattgt taacaatgta tttggtgcaa tttcgtcacg
     2701 taaattttta atagcttcga ttaattccca atgtgcatct actttactca tttctttacg
     2761 tgtacgaaga tgaatagata aattggcaat gtcttgttcg aagacgtgct tcaaccaatc
     2821 tttccattca tcgatttcat agtagccaag gcgtgtttta acacttaccg gaagcccacc
     2881 tgctttagtc gcttgaataa tttcggcagc aacgtcaggt cttaagatta agccggaacc
     2941 cttacggtac ccggggatcc tctagagtcg acctgcaggc atgcaagctc tgatttcact
     3001 ttttgcattc tacaaactgc ataactcata tgtaaatcgc tcctttttag gtggcacaaa
     3061 tgtgaggcat tttcgctctt tccggcgagg ctagttaccc ttaagttatt ggtatgactg
     3121 gttttaagcg caaaaaaagt tgctttttcg tacctattaa tgtatcgtta gaaaaccgac
     3181 tgtaaaaagt acagtcggca ttatctcata ttataaaagc cagtcattag gcctatctga
     3241 caattcctga atagagttca taaacaatcc tgcatgataa ccatcacaaa cagaatgatg
     3301 tacctgtaaa gatagcggta aatatattga attaccttta ttaatgaatt ttcctgctgt
     3361 aataatgggt agaaggtaat tactattatt attgatattt aagttaaacc cagtaaatga
     3421 agtccatgga ataatagaaa gagaaaaagc attttcaggt ataggtgttt tgggaaacaa
     3481 tttccccgaa ccattatatt tctctacatc agaaaggtat aaatcataaa actctttgaa
     3541 gtcattcttt acaggagtcc aaataccaga gaatgtttta gatacaccat caaaaattgt
     3601 ataaagtggc tctaacttat cccaataacc taactctccg tcgctattgt aaccagttct
     3661 aaaagctgta tttgagttta tcacccttgt cactaagaaa ataaatgcag ggtaaaattt
     3721 atatccttct tgttttatgt ttcggtataa aacactaata tcaatttctg tggttatact
     3781 aaaagtcgtt tgttggttca aataatgatt aaatatctct tttctcttcc aattgtctaa
     3841 atcaatttta ttaaagttca tttgatatgc ctcctaaatt tttatctaaa gtgaatttag
     3901 gaggcttact tgtctgcttt cttcattaga atcaatcctt ttttaaaagt caatattact
     3961 gtaacataaa tatatatttt aaaaatatcc cactttatcc aattttcgtt tgttgaacta
     4021 atgggtgctt tagttgaaga ataaagacca cattaaaaaa tgtggtcttt tgtgtttttt
     4081 taaaggattt gagcgtagcg aaaaatcctt ttctttctta tcttgataat aagggtaact
     4141 attgccggga tagactgtaa cattctcacg cataaaatcc cctttcattt tctaatgtaa
     4201 atctattacc ttattattaa ttcaattcgc tcataattaa tcctttttct tattacgcaa
     4261 aatggcccga tttaagcaca ccctttattc cgttaatgcg ccatgacagc catgataatt
     4321 actaatacta ggagaagtta ataaatacgg gtgatctaat gattcaatga ttcaaaccct
     4381 tgtgaacttc tttagaacaa aagaggttcg taacaagatt ttcttcacac tagcaatgtt
     4441 agtaattttt aaaataggga cttatatacc agctccagga gtaaatcctg cagcttttga
     4501 taatccccaa ggttctcaag gtgccactga gttattaaat acttttggtg gcggagcctt
     4561 gaaacgattt tctatttttg caatgggtat tgtaccctac atcactgcat caatcgtaat
     4621 gcaattatta caaatggata ttgtccctaa attctcagaa tgggcaaaac aaggtgaagt
     4681 aggtagaaga aagttaaata acgttactcg ttatttagca atttctttag catttatcca
     4741 atctataggt atggcattcc aatttaataa ttatctcaaa ggtgcgctga ttatcaatca
     4801 gtcaattatg agttatttat taatagcact agttttgaca gcaggaactg ctttcttaat
     4861 atggcttggt gatcaaatca ctcagttcgg tgttggtaat ggtatttcta ttatcatatt
     4921 cccatcaagc ttattttaat tatactctat caatgataga gtgtcaatat tttttttagt
     4981 ttttcatgaa ctcgatcgag gggatccaaa taaaaaacta gtttgacaaa taactctatc
     5041 aatgatagag tgtcaacaaa aaggaggaat taatgatgtc tagattagat aaaagtaaag
     5101 tgattaacag cgcattagag ctgcttaatg aggtcggaat cgaaggttta acaacccgta
     5161 aactcgccca gaagctaggt gtagagcagc ctacattgta ttggcatgta aaaaataagc
     5221 gggctttgct cgacgcctta gccattgaga tgttagatag gcaccatact cacttttgcc
     5281 ctttagaagg ggaaagctgg caagattttt tacgtaataa cgctaaaagt tttagatgtg
     5341 ctttactaag tcatcgcgat ggagcaaaag tacatttagg tacacggcct acagaaaaac
     5401 agtatgaaac tctcgaaaat caattagcct ttttatgcca acaaggtttt tcactagaga
     5461 atgcattata tgcactcagc gctgtggggc attttacttt aggttgcgta ttggaagatc
     5521 aagagcatca agtcgctaaa gaagaaaggg aaacacctac tactgatagt atgccgccat
     5581 tattacgaca agctatcgaa ttatttgatc accaaggtgc agagccagcc ttcttattcg
     5641 gccttgaatt gatcatatgc ggattagaaa aacaacttaa atgtgaaagt gggtcttaaa
     5701 agcagcataa cctttttccg tgatggtaac ttcacggtaa ccaagataac aaagaataca
     5761 agaaaatatt tacaaaaaat caatttaaca attccttaaa acatgcagga attgacgatt
     5821 taaacaatat tagctttgaa caattcttat ctcttttcaa tagctataaa ttatttaata
     5881 agtaagttaa gggatgcata aactgcatcc cttaacttgt ttttcgtgtg cctatttttt
     5941 gtgaatcgat tatgtctttt gcgcagtcgg cttaaaccag ttttcgctgg tgcgaaaaaa
     6001 gagtgtcttg tgacacctaa attcaaaatc tatcggtcag atttataccg atttgatttt
     6061 atatattctt gaataacata cgccgagtta tcacataaaa gcgggaacca atcatcaaat
     6121 ttaaacttca ttgcataatc cattaaactc ttaaattcta cgattccttg ttcatcaata
     6181 aactcaatca tttctttaat taatttatat ctatctgttg ttgttttctt taataattca
     6241 tcaacatcta caccgccata aactatcata tcttcttttt gatatttaaa tttattagga
     6301 tcgtccatgt gaagcatata tctcacaaga cctttcacac ttcctgcaat ctgcggaata
     6361 gtcgcattca attcttctgt aattattttt atctgttcat aagatttatt accctcatac
     6421 atcactagaa tatgataatg ctcttttttc atcctatctt ctgtatcagt atccctatca
     6481 tgtaatggag acactacaaa ttgaatgtgt aactctttta aatactctaa ccactcggct
     6541 tttgctgatt ctggatataa aacaaatgtc caattacgtc ctcttgaatt tttcttgttt
     6601 tcagtttctt ttattacatt ttcgctcatg atataataac ggtgctaata catttaacaa
     6661 aatttagtca tagataggca gcatgccagt gctgtctatc tttttttgtt taaaatgcac
     6721 cgtattcctc ctttgcatat ttttttatta gaataccggt tgcatctgat ttgctaatat
     6781 tatatttttc tttgattcta tttaatatct cattttcttc tgttgtaagt cttaaagtaa
     6841 cagcaacttt tttctcttct tttctatcta caaccatcac tgtacctccc aacatctgtt
     6901 tttttcactt taacataaaa aacaaccttt taacattaaa aacccaatat ttatttattt
     6961 gtttggacaa tggacaatgg acacctaggg gggaggtcgt agtacccccc tatgttttct
     7021 cccctaaata accccaaaaa tctaagaaaa aaagacctca aaaaggtctt taattaacat
     7081 ctcaaatttc gcatttattc caatttcctt tttgcgtgtg atgcgctgcg tccattaaaa
     7141 atcctagagc tttgcaaccg aaagttaata gctgtcgcta ctactttcgc ttacgctcta
     7201 agtatatttt aaggactgtc acacgcaaaa agttttctcg gcataaaagt acctctacat
     7261 ctctaaatcg tctgtacgct gtttctcacg ctttctatcg accttctgga cattatcctg
     7321 tacaacatcc ataaactgtc ccacacgctc gaatttggaa tcattaaaga atttctcttt
     7381 aagcctatta aaccctttct caaacccagg gaaattcgcc ctcgcagcac gatataaagt
     7441 cactgtacta tcttgaaatt tctctgatac attcaactgc tcattcaaac tatcattctc
     7501 tcgctttaat ttattaacct ctttactttt ttcgtgatac ccctctttcc atgtattcac
     7561 tacttctttc aaactctctc tacgtttttt taattcttga ttttctgtgt aatagtctgt
     7621 gctcttaata ttttcgtaat catcaacaat ccgttcctgc ctcgcgcgtt tcggtgatga
     7681 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     7741 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     7801 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
     7861 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     7921 agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     7981 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     8041 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     8101 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     8161 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     8221 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     8281 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     8341 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     8401 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     8461 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     8521 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     8581 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     8641 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     8701 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     8761 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     8821 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     8881 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     8941 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     9001 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     9061 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     9121 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     9181 aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     9241 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     9301 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     9361 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     9421 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     9481 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     9541 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     9601 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     9661 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     9721 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     9781 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     9841 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     9901 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaa
//
