LOCUS       sequence-392397-        9939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pJB38-NWMN29-30 + SarA_P1-mCherry-Term.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9939)
  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 9939)
  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: pRN111
FEATURES             Location/Qualifiers
     source          1..9939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2049..2065)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2049..2065)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      2074..2108
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     protein_bind    4935..4953
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    5024..5042
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             5064..5690
                     /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(5140..5159)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(7681..7703)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    7789..7929
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7844..7863
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7944..7961)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8115..8703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8195..8214)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8874..9665,9666..9734))
                     /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     9497..9516
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9735..9839)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9907..9925
                     /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 aggatgtata
     1321 catatggtga gcaagggcga ggaggataat atggctatta tcaaagaatt tatgagattc
     1381 aaagttcata tggaaggttc agtcaatggt cacgaatttg aaattgaggg agaaggtgaa
     1441 ggcagaccat atgaaggtac tcaaacggcc aaacttaaag tcactaaagg aggtcctctt
     1501 ccatttgcat gggatattct tagtccacaa tttatgtatg gatctaaagc ttacgttaaa
     1561 catccagcgg acattccgga ttatcttaaa ttgtcgtttc cagaaggctt caaatgggaa
     1621 agagtaatga actttgaaga tggaggagtc gttacagtca ctcaggatag ttctcttcaa
     1681 gatggagaat ttatctacaa agtcaagtta agaggtacca attttccctc agatggtcca
     1741 gttatgcaaa agaaaactat gggatgggaa gcatccagtg agcgtatgta tcctgaagat
     1801 ggtgccctta aaggcgaaat taaacaacgc ctaaagctta aagatggtgg acattatgac
     1861 gctgaagtta aaacaactta caaagcgaag aaaccggtac aacttccagg tgcatataat
     1921 gtcaacatta aacttgatat tacaagtcat aatgaagatt atactattgt tgaacaatat
     1981 gagagagctg aaggaagaca ttctacaggc ggcatggacg agctgtacaa gtaagaattc
     2041 gtaatcatgt catagctgtt tcctgtgtgt taattcagaa cgctcggttg ccgccgggcg
     2101 ttttttatgc agctcgcgat atatcctgat gtatttttaa ttttgcacca tttccaaaag
     2161 gataagtgac ataagtaaaa aggcatcatc gggagttatc ctatcaggaa aaccaagata
     2221 atacctaagt agaaagtgtt caatccgtgt taaattggga aatatcatcc ataaacttta
     2281 ttactcatac tataattcaa ttttaacgtc ttcgtccatt tgggcttcaa attcatcgag
     2341 tagtgctcgt gcttctgcaa ttgattgtgt gttcatcaat tgatgtcgaa gttcgctagc
     2401 gcctcttatg ccacgcacat agattttaaa gaatctacgc aagctcttga attgtcgtat
     2461 ttcatctttt tcatatttgt taaacaatga taaatgcaat ctcaatagat ctaatagttc
     2521 cttgcttgtg tgttcgcgtg gttctttttc aaaagcgaat ggattgtgga aaatgcctct
     2581 accaatcatg acgccatcaa tgccatattt ttctgccagt tcaagtcctg tttttctatc
     2641 gggaatatca ccgttaattg ttaacaatgt atttggtgca atttcgtcac gtaaattttt
     2701 aatagcttcg attaattccc aatgtgcatc tactttactc atttctttac gtgtacgaag
     2761 atgaatagat aaattggcaa tgtcttgttc gaagacgtgc ttcaaccaat ctttccattc
     2821 atcgatttca tagtagccaa ggcgtgtttt aacacttacc ggaagcccac ctgctttagt
     2881 cgcttgaata atttcggcag caacgtcagg tcttaagatt aagccggaac ccttacggta
     2941 cccggggatc ctctagagtc gacctgcagg catgcaagct ctgatttcac tttttgcatt
     3001 ctacaaactg cataactcat atgtaaatcg ctccttttta ggtggcacaa atgtgaggca
     3061 ttttcgctct ttccggcgag gctagttacc cttaagttat tggtatgact ggttttaagc
     3121 gcaaaaaaag ttgctttttc gtacctatta atgtatcgtt agaaaaccga ctgtaaaaag
     3181 tacagtcggc attatctcat attataaaag ccagtcatta ggcctatctg acaattcctg
     3241 aatagagttc ataaacaatc ctgcatgata accatcacaa acagaatgat gtacctgtaa
     3301 agatagcggt aaatatattg aattaccttt attaatgaat tttcctgctg taataatggg
     3361 tagaaggtaa ttactattat tattgatatt taagttaaac ccagtaaatg aagtccatgg
     3421 aataatagaa agagaaaaag cattttcagg tataggtgtt ttgggaaaca atttccccga
     3481 accattatat ttctctacat cagaaaggta taaatcataa aactctttga agtcattctt
     3541 tacaggagtc caaataccag agaatgtttt agatacacca tcaaaaattg tataaagtgg
     3601 ctctaactta tcccaataac ctaactctcc gtcgctattg taaccagttc taaaagctgt
     3661 atttgagttt atcacccttg tcactaagaa aataaatgca gggtaaaatt tatatccttc
     3721 ttgttttatg tttcggtata aaacactaat atcaatttct gtggttatac taaaagtcgt
     3781 ttgttggttc aaataatgat taaatatctc ttttctcttc caattgtcta aatcaatttt
     3841 attaaagttc atttgatatg cctcctaaat ttttatctaa agtgaattta ggaggcttac
     3901 ttgtctgctt tcttcattag aatcaatcct tttttaaaag tcaatattac tgtaacataa
     3961 atatatattt taaaaatatc ccactttatc caattttcgt ttgttgaact aatgggtgct
     4021 ttagttgaag aataaagacc acattaaaaa atgtggtctt ttgtgttttt ttaaaggatt
     4081 tgagcgtagc gaaaaatcct tttctttctt atcttgataa taagggtaac tattgccggg
     4141 atagactgta acattctcac gcataaaatc ccctttcatt ttctaatgta aatctattac
     4201 cttattatta attcaattcg ctcataatta atcctttttc ttattacgca aaatggcccg
     4261 atttaagcac accctttatt ccgttaatgc gccatgacag ccatgataat tactaatact
     4321 aggagaagtt aataaatacg ggtgatctaa tgattcaatg attcaaaccc ttgtgaactt
     4381 ctttagaaca aaagaggttc gtaacaagat tttcttcaca ctagcaatgt tagtaatttt
     4441 taaaataggg acttatatac cagctccagg agtaaatcct gcagcttttg ataatcccca
     4501 aggttctcaa ggtgccactg agttattaaa tacttttggt ggcggagcct tgaaacgatt
     4561 ttctattttt gcaatgggta ttgtacccta catcactgca tcaatcgtaa tgcaattatt
     4621 acaaatggat attgtcccta aattctcaga atgggcaaaa caaggtgaag taggtagaag
     4681 aaagttaaat aacgttactc gttatttagc aatttcttta gcatttatcc aatctatagg
     4741 tatggcattc caatttaata attatctcaa aggtgcgctg attatcaatc agtcaattat
     4801 gagttattta ttaatagcac tagttttgac agcaggaact gctttcttaa tatggcttgg
     4861 tgatcaaatc actcagttcg gtgttggtaa tggtatttct attatcatat tcccatcaag
     4921 cttattttaa ttatactcta tcaatgatag agtgtcaata ttttttttag tttttcatga
     4981 actcgatcga ggggatccaa ataaaaaact agtttgacaa ataactctat caatgataga
     5041 gtgtcaacaa aaaggaggaa ttaatgatgt ctagattaga taaaagtaaa gtgattaaca
     5101 gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt aacaacccgt aaactcgccc
     5161 agaagctagg tgtagagcag cctacattgt attggcatgt aaaaaataag cgggctttgc
     5221 tcgacgcctt agccattgag atgttagata ggcaccatac tcacttttgc cctttagaag
     5281 gggaaagctg gcaagatttt ttacgtaata acgctaaaag ttttagatgt gctttactaa
     5341 gtcatcgcga tggagcaaaa gtacatttag gtacacggcc tacagaaaaa cagtatgaaa
     5401 ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt ttcactagag aatgcattat
     5461 atgcactcag cgctgtgggg cattttactt taggttgcgt attggaagat caagagcatc
     5521 aagtcgctaa agaagaaagg gaaacaccta ctactgatag tatgccgcca ttattacgac
     5581 aagctatcga attatttgat caccaaggtg cagagccagc cttcttattc ggccttgaat
     5641 tgatcatatg cggattagaa aaacaactta aatgtgaaag tgggtcttaa aagcagcata
     5701 acctttttcc gtgatggtaa cttcacggta accaagataa caaagaatac aagaaaatat
     5761 ttacaaaaaa tcaatttaac aattccttaa aacatgcagg aattgacgat ttaaacaata
     5821 ttagctttga acaattctta tctcttttca atagctataa attatttaat aagtaagtta
     5881 agggatgcat aaactgcatc ccttaacttg tttttcgtgt gcctattttt tgtgaatcga
     5941 ttatgtcttt tgcgcagtcg gcttaaacca gttttcgctg gtgcgaaaaa agagtgtctt
     6001 gtgacaccta aattcaaaat ctatcggtca gatttatacc gatttgattt tatatattct
     6061 tgaataacat acgccgagtt atcacataaa agcgggaacc aatcatcaaa tttaaacttc
     6121 attgcataat ccattaaact cttaaattct acgattcctt gttcatcaat aaactcaatc
     6181 atttctttaa ttaatttata tctatctgtt gttgttttct ttaataattc atcaacatct
     6241 acaccgccat aaactatcat atcttctttt tgatatttaa atttattagg atcgtccatg
     6301 tgaagcatat atctcacaag acctttcaca cttcctgcaa tctgcggaat agtcgcattc
     6361 aattcttctg taattatttt tatctgttca taagatttat taccctcata catcactaga
     6421 atatgataat gctctttttt catcctatct tctgtatcag tatccctatc atgtaatgga
     6481 gacactacaa attgaatgtg taactctttt aaatactcta accactcggc ttttgctgat
     6541 tctggatata aaacaaatgt ccaattacgt cctcttgaat ttttcttgtt ttcagtttct
     6601 tttattacat tttcgctcat gatataataa cggtgctaat acatttaaca aaatttagtc
     6661 atagataggc agcatgccag tgctgtctat ctttttttgt ttaaaatgca ccgtattcct
     6721 cctttgcata tttttttatt agaataccgg ttgcatctga tttgctaata ttatattttt
     6781 ctttgattct atttaatatc tcattttctt ctgttgtaag tcttaaagta acagcaactt
     6841 ttttctcttc ttttctatct acaaccatca ctgtacctcc caacatctgt ttttttcact
     6901 ttaacataaa aaacaacctt ttaacattaa aaacccaata tttatttatt tgtttggaca
     6961 atggacaatg gacacctagg ggggaggtcg tagtaccccc ctatgttttc tcccctaaat
     7021 aaccccaaaa atctaagaaa aaaagacctc aaaaaggtct ttaattaaca tctcaaattt
     7081 cgcatttatt ccaatttcct ttttgcgtgt gatgcgctgc gtccattaaa aatcctagag
     7141 ctttgcaacc gaaagttaat agctgtcgct actactttcg cttacgctct aagtatattt
     7201 taaggactgt cacacgcaaa aagttttctc ggcataaaag tacctctaca tctctaaatc
     7261 gtctgtacgc tgtttctcac gctttctatc gaccttctgg acattatcct gtacaacatc
     7321 cataaactgt cccacacgct cgaatttgga atcattaaag aatttctctt taagcctatt
     7381 aaaccctttc tcaaacccag ggaaattcgc cctcgcagca cgatataaag tcactgtact
     7441 atcttgaaat ttctctgata cattcaactg ctcattcaaa ctatcattct ctcgctttaa
     7501 tttattaacc tctttacttt tttcgtgata cccctctttc catgtattca ctacttcttt
     7561 caaactctct ctacgttttt ttaattcttg attttctgtg taatagtctg tgctcttaat
     7621 attttcgtaa tcatcaacaa tccgttcctg cctcgcgcgt ttcggtgatg acggtgaaaa
     7681 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     7741 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     7801 ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt
     7861 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     7921 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7981 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     8041 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     8101 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     8161 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     8221 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     8281 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     8341 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     8401 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     8461 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     8521 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     8581 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     8641 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     8701 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     8761 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     8821 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     8881 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     8941 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     9001 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     9061 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     9121 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     9181 gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     9241 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     9301 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     9361 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     9421 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     9481 ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     9541 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     9601 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     9661 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     9721 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     9781 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     9841 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     9901 tataaaaata ggcgtatcac gaggcccttt cgtcttcaa
//
