LOCUS       sequence-168588-        9924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pJB38-NWMN29-30 + SarA_P1-mKate2-Term.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9924)
  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 9924)
  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: pRN110
FEATURES             Location/Qualifiers
     source          1..9924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2034..2050)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2034..2050)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      2059..2093
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     protein_bind    4920..4938
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    5009..5027
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             5049..5675
                     /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(5125..5144)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(7666..7688)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    7774..7914
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7829..7848
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7929..7946)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8100..8688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8180..8199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8859..9650,9651..9719))
                     /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     9482..9501
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9720..9824)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9892..9910
                     /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 tttatgtcag aacttatcaa ggaaaatatg cacatgaaat tgtacatgga aggaacagta
     1381 aataatcacc actttaaatg tacctcagaa ggagaaggaa aaccatatga aggtactcaa
     1441 accatgcgta ttaaggccgt tgaaggtgga ccattgcctt ttgcctttga tattcttgct
     1501 acatctttta tgtacggatc aaagactttt atcaatcata cccaaggtat cccagatttc
     1561 tttaaacagt catttcctga aggatttaca tgggaacgtg tcacaactta tgaagatggt
     1621 ggagtattga cagcaactca agatacatct ttgcaagatg gttgtcttat ctacaatgta
     1681 aagatccgtg gagttaattt tccaagtaat ggtcctgtta tgcagaaaaa gacccttgga
     1741 tgggaagcat ctaccgaaac attatatcct gccgatggtg gattggaagg tcgtgctgat
     1801 atggcattga aacttgtcgg tggaggtcac cttatctgta atttgaagac cacataccgt
     1861 tctaaaaagc cagctaagaa tcttaagatg cctggtgttt actacgtcga tcgtcgttta
     1921 gaacgtatca aagaagcaga taaggaaact tatgttgaac agcacgaagt agccgtcgca
     1981 cgttattgtg atttgcctag taaattggga caccgttaag aattcgtaat catgtcatag
     2041 ctgtttcctg tgtgttaatt cagaacgctc ggttgccgcc gggcgttttt tatgcagctc
     2101 gcgatatatc ctgatgtatt tttaattttg caccatttcc aaaaggataa gtgacataag
     2161 taaaaaggca tcatcgggag ttatcctatc aggaaaacca agataatacc taagtagaaa
     2221 gtgttcaatc cgtgttaaat tgggaaatat catccataaa ctttattact catactataa
     2281 ttcaatttta acgtcttcgt ccatttgggc ttcaaattca tcgagtagtg ctcgtgcttc
     2341 tgcaattgat tgtgtgttca tcaattgatg tcgaagttcg ctagcgcctc ttatgccacg
     2401 cacatagatt ttaaagaatc tacgcaagct cttgaattgt cgtatttcat ctttttcata
     2461 tttgttaaac aatgataaat gcaatctcaa tagatctaat agttccttgc ttgtgtgttc
     2521 gcgtggttct ttttcaaaag cgaatggatt gtggaaaatg cctctaccaa tcatgacgcc
     2581 atcaatgcca tatttttctg ccagttcaag tcctgttttt ctatcgggaa tatcaccgtt
     2641 aattgttaac aatgtatttg gtgcaatttc gtcacgtaaa tttttaatag cttcgattaa
     2701 ttcccaatgt gcatctactt tactcatttc tttacgtgta cgaagatgaa tagataaatt
     2761 ggcaatgtct tgttcgaaga cgtgcttcaa ccaatctttc cattcatcga tttcatagta
     2821 gccaaggcgt gttttaacac ttaccggaag cccacctgct ttagtcgctt gaataatttc
     2881 ggcagcaacg tcaggtctta agattaagcc ggaaccctta cggtacccgg ggatcctcta
     2941 gagtcgacct gcaggcatgc aagctctgat ttcacttttt gcattctaca aactgcataa
     3001 ctcatatgta aatcgctcct ttttaggtgg cacaaatgtg aggcattttc gctctttccg
     3061 gcgaggctag ttacccttaa gttattggta tgactggttt taagcgcaaa aaaagttgct
     3121 ttttcgtacc tattaatgta tcgttagaaa accgactgta aaaagtacag tcggcattat
     3181 ctcatattat aaaagccagt cattaggcct atctgacaat tcctgaatag agttcataaa
     3241 caatcctgca tgataaccat cacaaacaga atgatgtacc tgtaaagata gcggtaaata
     3301 tattgaatta cctttattaa tgaattttcc tgctgtaata atgggtagaa ggtaattact
     3361 attattattg atatttaagt taaacccagt aaatgaagtc catggaataa tagaaagaga
     3421 aaaagcattt tcaggtatag gtgttttggg aaacaatttc cccgaaccat tatatttctc
     3481 tacatcagaa aggtataaat cataaaactc tttgaagtca ttctttacag gagtccaaat
     3541 accagagaat gttttagata caccatcaaa aattgtataa agtggctcta acttatccca
     3601 ataacctaac tctccgtcgc tattgtaacc agttctaaaa gctgtatttg agtttatcac
     3661 ccttgtcact aagaaaataa atgcagggta aaatttatat ccttcttgtt ttatgtttcg
     3721 gtataaaaca ctaatatcaa tttctgtggt tatactaaaa gtcgtttgtt ggttcaaata
     3781 atgattaaat atctcttttc tcttccaatt gtctaaatca attttattaa agttcatttg
     3841 atatgcctcc taaattttta tctaaagtga atttaggagg cttacttgtc tgctttcttc
     3901 attagaatca atcctttttt aaaagtcaat attactgtaa cataaatata tattttaaaa
     3961 atatcccact ttatccaatt ttcgtttgtt gaactaatgg gtgctttagt tgaagaataa
     4021 agaccacatt aaaaaatgtg gtcttttgtg tttttttaaa ggatttgagc gtagcgaaaa
     4081 atccttttct ttcttatctt gataataagg gtaactattg ccgggataga ctgtaacatt
     4141 ctcacgcata aaatcccctt tcattttcta atgtaaatct attaccttat tattaattca
     4201 attcgctcat aattaatcct ttttcttatt acgcaaaatg gcccgattta agcacaccct
     4261 ttattccgtt aatgcgccat gacagccatg ataattacta atactaggag aagttaataa
     4321 atacgggtga tctaatgatt caatgattca aacccttgtg aacttcttta gaacaaaaga
     4381 ggttcgtaac aagattttct tcacactagc aatgttagta atttttaaaa tagggactta
     4441 tataccagct ccaggagtaa atcctgcagc ttttgataat ccccaaggtt ctcaaggtgc
     4501 cactgagtta ttaaatactt ttggtggcgg agccttgaaa cgattttcta tttttgcaat
     4561 gggtattgta ccctacatca ctgcatcaat cgtaatgcaa ttattacaaa tggatattgt
     4621 ccctaaattc tcagaatggg caaaacaagg tgaagtaggt agaagaaagt taaataacgt
     4681 tactcgttat ttagcaattt ctttagcatt tatccaatct ataggtatgg cattccaatt
     4741 taataattat ctcaaaggtg cgctgattat caatcagtca attatgagtt atttattaat
     4801 agcactagtt ttgacagcag gaactgcttt cttaatatgg cttggtgatc aaatcactca
     4861 gttcggtgtt ggtaatggta tttctattat catattccca tcaagcttat tttaattata
     4921 ctctatcaat gatagagtgt caatattttt tttagttttt catgaactcg atcgagggga
     4981 tccaaataaa aaactagttt gacaaataac tctatcaatg atagagtgtc aacaaaaagg
     5041 aggaattaat gatgtctaga ttagataaaa gtaaagtgat taacagcgca ttagagctgc
     5101 ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag
     5161 agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac gccttagcca
     5221 ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa agctggcaag
     5281 attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat cgcgatggag
     5341 caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc gaaaatcaat
     5401 tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca ctcagcgctg
     5461 tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag
     5521 aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct atcgaattat
     5581 ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc atatgcggat
     5641 tagaaaaaca acttaaatgt gaaagtgggt cttaaaagca gcataacctt tttccgtgat
     5701 ggtaacttca cggtaaccaa gataacaaag aatacaagaa aatatttaca aaaaatcaat
     5761 ttaacaattc cttaaaacat gcaggaattg acgatttaaa caatattagc tttgaacaat
     5821 tcttatctct tttcaatagc tataaattat ttaataagta agttaaggga tgcataaact
     5881 gcatccctta acttgttttt cgtgtgccta ttttttgtga atcgattatg tcttttgcgc
     5941 agtcggctta aaccagtttt cgctggtgcg aaaaaagagt gtcttgtgac acctaaattc
     6001 aaaatctatc ggtcagattt ataccgattt gattttatat attcttgaat aacatacgcc
     6061 gagttatcac ataaaagcgg gaaccaatca tcaaatttaa acttcattgc ataatccatt
     6121 aaactcttaa attctacgat tccttgttca tcaataaact caatcatttc tttaattaat
     6181 ttatatctat ctgttgttgt tttctttaat aattcatcaa catctacacc gccataaact
     6241 atcatatctt ctttttgata tttaaattta ttaggatcgt ccatgtgaag catatatctc
     6301 acaagacctt tcacacttcc tgcaatctgc ggaatagtcg cattcaattc ttctgtaatt
     6361 atttttatct gttcataaga tttattaccc tcatacatca ctagaatatg ataatgctct
     6421 tttttcatcc tatcttctgt atcagtatcc ctatcatgta atggagacac tacaaattga
     6481 atgtgtaact cttttaaata ctctaaccac tcggcttttg ctgattctgg atataaaaca
     6541 aatgtccaat tacgtcctct tgaatttttc ttgttttcag tttcttttat tacattttcg
     6601 ctcatgatat aataacggtg ctaatacatt taacaaaatt tagtcataga taggcagcat
     6661 gccagtgctg tctatctttt tttgtttaaa atgcaccgta ttcctccttt gcatattttt
     6721 ttattagaat accggttgca tctgatttgc taatattata tttttctttg attctattta
     6781 atatctcatt ttcttctgtt gtaagtctta aagtaacagc aacttttttc tcttcttttc
     6841 tatctacaac catcactgta cctcccaaca tctgtttttt tcactttaac ataaaaaaca
     6901 accttttaac attaaaaacc caatatttat ttatttgttt ggacaatgga caatggacac
     6961 ctagggggga ggtcgtagta cccccctatg ttttctcccc taaataaccc caaaaatcta
     7021 agaaaaaaag acctcaaaaa ggtctttaat taacatctca aatttcgcat ttattccaat
     7081 ttcctttttg cgtgtgatgc gctgcgtcca ttaaaaatcc tagagctttg caaccgaaag
     7141 ttaatagctg tcgctactac tttcgcttac gctctaagta tattttaagg actgtcacac
     7201 gcaaaaagtt ttctcggcat aaaagtacct ctacatctct aaatcgtctg tacgctgttt
     7261 ctcacgcttt ctatcgacct tctggacatt atcctgtaca acatccataa actgtcccac
     7321 acgctcgaat ttggaatcat taaagaattt ctctttaagc ctattaaacc ctttctcaaa
     7381 cccagggaaa ttcgccctcg cagcacgata taaagtcact gtactatctt gaaatttctc
     7441 tgatacattc aactgctcat tcaaactatc attctctcgc tttaatttat taacctcttt
     7501 acttttttcg tgatacccct ctttccatgt attcactact tctttcaaac tctctctacg
     7561 tttttttaat tcttgatttt ctgtgtaata gtctgtgctc ttaatatttt cgtaatcatc
     7621 aacaatccgt tcctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     7681 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     7741 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     7801 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     7861 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct
     7921 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     7981 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     8041 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     8101 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     8161 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     8221 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     8281 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     8341 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     8401 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     8461 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     8521 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     8581 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     8641 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     8701 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     8761 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     8821 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     8881 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     8941 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     9001 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     9061 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     9121 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc
     9181 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     9241 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     9301 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     9361 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     9421 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg
     9481 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     9541 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     9601 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     9661 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     9721 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     9781 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     9841 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     9901 atcacgaggc cctttcgtct tcaa
//
