LOCUS       sequence-163697-        9902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pJB38-NWMN29-30 + SarA_P1-DsRed-Term.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9902)
  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 9902)
  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: pTH101
FEATURES             Location/Qualifiers
     source          1..9902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1320..1997
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MDNTEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEPSTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(1450..1468)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(1500..1520)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1718..1737
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1912..1935
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2012..2028)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2012..2028)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      2037..2071
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     protein_bind    4898..4916
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    4987..5005
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             5027..5653
                     /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(5103..5122)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(7644..7666)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    7752..7892
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7807..7826
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7907..7924)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8078..8666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8158..8177)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8837..9628,9629..9697))
                     /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     9460..9479
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9698..9802)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9870..9888
                     /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 gtaccaggag gaaaaacata
     1321 tggacaacac cgaggacgtc atcaaggagt tcatgcgctt caaggtgcgc atggagggct
     1381 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     1441 cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc
     1501 tgtcccccca gttccagtac ggctccaagg tgtacgtgaa gcaccccgcc gacatccccg
     1561 actacaagaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     1621 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggctgc ttcatctaca
     1681 aggtgaagtt catcggcgtg aacttcccct ccgacggccc cgtaatgcag aagaagacta
     1741 tgggctggga gccctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga
     1801 tccacaaggc cctgaagctg aaggacggcg gccactacct ggtggagttc aagtccatct
     1861 acatggccaa gaagcccgtg cagctgcccg gctactacta cgtggactcc aagctggaca
     1921 tcacctccca caacgaggac tacaccatcg tggagcagta cgagcgcacc gagggccgcc
     1981 accacctgtt cctgtaggaa ttcgtaatca tgtcatagct gtttcctgtg tgttaattca
     2041 gaacgctcgg ttgccgccgg gcgtttttta tgcagctcgc gatatatcct gatgtatttt
     2101 taattttgca ccatttccaa aaggataagt gacataagta aaaaggcatc atcgggagtt
     2161 atcctatcag gaaaaccaag ataataccta agtagaaagt gttcaatccg tgttaaattg
     2221 ggaaatatca tccataaact ttattactca tactataatt caattttaac gtcttcgtcc
     2281 atttgggctt caaattcatc gagtagtgct cgtgcttctg caattgattg tgtgttcatc
     2341 aattgatgtc gaagttcgct agcgcctctt atgccacgca catagatttt aaagaatcta
     2401 cgcaagctct tgaattgtcg tatttcatct ttttcatatt tgttaaacaa tgataaatgc
     2461 aatctcaata gatctaatag ttccttgctt gtgtgttcgc gtggttcttt ttcaaaagcg
     2521 aatggattgt ggaaaatgcc tctaccaatc atgacgccat caatgccata tttttctgcc
     2581 agttcaagtc ctgtttttct atcgggaata tcaccgttaa ttgttaacaa tgtatttggt
     2641 gcaatttcgt cacgtaaatt tttaatagct tcgattaatt cccaatgtgc atctacttta
     2701 ctcatttctt tacgtgtacg aagatgaata gataaattgg caatgtcttg ttcgaagacg
     2761 tgcttcaacc aatctttcca ttcatcgatt tcatagtagc caaggcgtgt tttaacactt
     2821 accggaagcc cacctgcttt agtcgcttga ataatttcgg cagcaacgtc aggtcttaag
     2881 attaagccgg aacccttacg gtacccgggg atcctctaga gtcgacctgc aggcatgcaa
     2941 gctctgattt cactttttgc attctacaaa ctgcataact catatgtaaa tcgctccttt
     3001 ttaggtggca caaatgtgag gcattttcgc tctttccggc gaggctagtt acccttaagt
     3061 tattggtatg actggtttta agcgcaaaaa aagttgcttt ttcgtaccta ttaatgtatc
     3121 gttagaaaac cgactgtaaa aagtacagtc ggcattatct catattataa aagccagtca
     3181 ttaggcctat ctgacaattc ctgaatagag ttcataaaca atcctgcatg ataaccatca
     3241 caaacagaat gatgtacctg taaagatagc ggtaaatata ttgaattacc tttattaatg
     3301 aattttcctg ctgtaataat gggtagaagg taattactat tattattgat atttaagtta
     3361 aacccagtaa atgaagtcca tggaataata gaaagagaaa aagcattttc aggtataggt
     3421 gttttgggaa acaatttccc cgaaccatta tatttctcta catcagaaag gtataaatca
     3481 taaaactctt tgaagtcatt ctttacagga gtccaaatac cagagaatgt tttagataca
     3541 ccatcaaaaa ttgtataaag tggctctaac ttatcccaat aacctaactc tccgtcgcta
     3601 ttgtaaccag ttctaaaagc tgtatttgag tttatcaccc ttgtcactaa gaaaataaat
     3661 gcagggtaaa atttatatcc ttcttgtttt atgtttcggt ataaaacact aatatcaatt
     3721 tctgtggtta tactaaaagt cgtttgttgg ttcaaataat gattaaatat ctcttttctc
     3781 ttccaattgt ctaaatcaat tttattaaag ttcatttgat atgcctccta aatttttatc
     3841 taaagtgaat ttaggaggct tacttgtctg ctttcttcat tagaatcaat ccttttttaa
     3901 aagtcaatat tactgtaaca taaatatata ttttaaaaat atcccacttt atccaatttt
     3961 cgtttgttga actaatgggt gctttagttg aagaataaag accacattaa aaaatgtggt
     4021 cttttgtgtt tttttaaagg atttgagcgt agcgaaaaat ccttttcttt cttatcttga
     4081 taataagggt aactattgcc gggatagact gtaacattct cacgcataaa atcccctttc
     4141 attttctaat gtaaatctat taccttatta ttaattcaat tcgctcataa ttaatccttt
     4201 ttcttattac gcaaaatggc ccgatttaag cacacccttt attccgttaa tgcgccatga
     4261 cagccatgat aattactaat actaggagaa gttaataaat acgggtgatc taatgattca
     4321 atgattcaaa cccttgtgaa cttctttaga acaaaagagg ttcgtaacaa gattttcttc
     4381 acactagcaa tgttagtaat ttttaaaata gggacttata taccagctcc aggagtaaat
     4441 cctgcagctt ttgataatcc ccaaggttct caaggtgcca ctgagttatt aaatactttt
     4501 ggtggcggag ccttgaaacg attttctatt tttgcaatgg gtattgtacc ctacatcact
     4561 gcatcaatcg taatgcaatt attacaaatg gatattgtcc ctaaattctc agaatgggca
     4621 aaacaaggtg aagtaggtag aagaaagtta aataacgtta ctcgttattt agcaatttct
     4681 ttagcattta tccaatctat aggtatggca ttccaattta ataattatct caaaggtgcg
     4741 ctgattatca atcagtcaat tatgagttat ttattaatag cactagtttt gacagcagga
     4801 actgctttct taatatggct tggtgatcaa atcactcagt tcggtgttgg taatggtatt
     4861 tctattatca tattcccatc aagcttattt taattatact ctatcaatga tagagtgtca
     4921 atattttttt tagtttttca tgaactcgat cgaggggatc caaataaaaa actagtttga
     4981 caaataactc tatcaatgat agagtgtcaa caaaaaggag gaattaatga tgtctagatt
     5041 agataaaagt aaagtgatta acagcgcatt agagctgctt aatgaggtcg gaatcgaagg
     5101 tttaacaacc cgtaaactcg cccagaagct aggtgtagag cagcctacat tgtattggca
     5161 tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt gagatgttag ataggcacca
     5221 tactcacttt tgccctttag aaggggaaag ctggcaagat tttttacgta ataacgctaa
     5281 aagttttaga tgtgctttac taagtcatcg cgatggagca aaagtacatt taggtacacg
     5341 gcctacagaa aaacagtatg aaactctcga aaatcaatta gcctttttat gccaacaagg
     5401 tttttcacta gagaatgcat tatatgcact cagcgctgtg gggcatttta ctttaggttg
     5461 cgtattggaa gatcaagagc atcaagtcgc taaagaagaa agggaaacac ctactactga
     5521 tagtatgccg ccattattac gacaagctat cgaattattt gatcaccaag gtgcagagcc
     5581 agccttctta ttcggccttg aattgatcat atgcggatta gaaaaacaac ttaaatgtga
     5641 aagtgggtct taaaagcagc ataacctttt tccgtgatgg taacttcacg gtaaccaaga
     5701 taacaaagaa tacaagaaaa tatttacaaa aaatcaattt aacaattcct taaaacatgc
     5761 aggaattgac gatttaaaca atattagctt tgaacaattc ttatctcttt tcaatagcta
     5821 taaattattt aataagtaag ttaagggatg cataaactgc atcccttaac ttgtttttcg
     5881 tgtgcctatt ttttgtgaat cgattatgtc ttttgcgcag tcggcttaaa ccagttttcg
     5941 ctggtgcgaa aaaagagtgt cttgtgacac ctaaattcaa aatctatcgg tcagatttat
     6001 accgatttga ttttatatat tcttgaataa catacgccga gttatcacat aaaagcggga
     6061 accaatcatc aaatttaaac ttcattgcat aatccattaa actcttaaat tctacgattc
     6121 cttgttcatc aataaactca atcatttctt taattaattt atatctatct gttgttgttt
     6181 tctttaataa ttcatcaaca tctacaccgc cataaactat catatcttct ttttgatatt
     6241 taaatttatt aggatcgtcc atgtgaagca tatatctcac aagacctttc acacttcctg
     6301 caatctgcgg aatagtcgca ttcaattctt ctgtaattat ttttatctgt tcataagatt
     6361 tattaccctc atacatcact agaatatgat aatgctcttt tttcatccta tcttctgtat
     6421 cagtatccct atcatgtaat ggagacacta caaattgaat gtgtaactct tttaaatact
     6481 ctaaccactc ggcttttgct gattctggat ataaaacaaa tgtccaatta cgtcctcttg
     6541 aatttttctt gttttcagtt tcttttatta cattttcgct catgatataa taacggtgct
     6601 aatacattta acaaaattta gtcatagata ggcagcatgc cagtgctgtc tatctttttt
     6661 tgtttaaaat gcaccgtatt cctcctttgc atattttttt attagaatac cggttgcatc
     6721 tgatttgcta atattatatt tttctttgat tctatttaat atctcatttt cttctgttgt
     6781 aagtcttaaa gtaacagcaa cttttttctc ttcttttcta tctacaacca tcactgtacc
     6841 tcccaacatc tgtttttttc actttaacat aaaaaacaac cttttaacat taaaaaccca
     6901 atatttattt atttgtttgg acaatggaca atggacacct aggggggagg tcgtagtacc
     6961 cccctatgtt ttctccccta aataacccca aaaatctaag aaaaaaagac ctcaaaaagg
     7021 tctttaatta acatctcaaa tttcgcattt attccaattt cctttttgcg tgtgatgcgc
     7081 tgcgtccatt aaaaatccta gagctttgca accgaaagtt aatagctgtc gctactactt
     7141 tcgcttacgc tctaagtata ttttaaggac tgtcacacgc aaaaagtttt ctcggcataa
     7201 aagtacctct acatctctaa atcgtctgta cgctgtttct cacgctttct atcgaccttc
     7261 tggacattat cctgtacaac atccataaac tgtcccacac gctcgaattt ggaatcatta
     7321 aagaatttct ctttaagcct attaaaccct ttctcaaacc cagggaaatt cgccctcgca
     7381 gcacgatata aagtcactgt actatcttga aatttctctg atacattcaa ctgctcattc
     7441 aaactatcat tctctcgctt taatttatta acctctttac ttttttcgtg atacccctct
     7501 ttccatgtat tcactacttc tttcaaactc tctctacgtt tttttaattc ttgattttct
     7561 gtgtaatagt ctgtgctctt aatattttcg taatcatcaa caatccgttc ctgcctcgcg
     7621 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     7681 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     7741 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
     7801 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     7861 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     7921 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     7981 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     8041 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     8101 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     8161 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     8221 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     8281 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     8341 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     8401 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     8461 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     8521 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     8581 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     8641 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     8701 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     8761 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     8821 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     8881 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     8941 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     9001 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     9061 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     9121 ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt
     9181 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     9241 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     9301 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     9361 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     9421 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
     9481 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     9541 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     9601 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     9661 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     9721 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     9781 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     9841 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     9901 aa
//
