LOCUS       sequence-163705-        9945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pJB38-NWMN29-30 + SarA_P1-YFP-Term.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9945)
  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 9945)
  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: pTH103
FEATURES             Location/Qualifiers
     source          1..9945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1354..1382)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1977..1997
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2055..2071)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2055..2071)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      2080..2114
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     protein_bind    4941..4959
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    5030..5048
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             5070..5696
                     /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(5146..5165)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(7687..7709)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    7795..7935
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7850..7869
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7950..7967)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8121..8709)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8201..8220)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8880..9671,9672..9740))
                     /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     9503..9522
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9741..9845)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9913..9931
                     /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 tttatgagta aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat
     1381 ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac
     1441 ggaaaactta cccttaaact tatttgcact actggaaaac tacctgttcc atggccaaca
     1501 cttgtcacta ctcttggata tggtcttcaa tgctttgcga gatacccaga tcatatgaaa
     1561 cagcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt
     1621 ttcaaagatg acgggaacta caagacacgt gctgaagtca agtttgaagg tgataccctt
     1681 gttaatagaa tcgagttaaa aggtattgat tttaaagaag atggaaacat tcttggacac
     1741 aaattggaat acaactataa ctcacacaat gtatacatca cagcagacaa acaaaagaat
     1801 ggaatcaaag ctaacttcaa aattagacac aacattgaag atggaggtgt tcaactagca
     1861 gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat
     1921 tacctgtcct atcaatctgc cctttcgaaa gatcccaacg aaaagagaga ccacatggtc
     1981 cttcttgagt ttgtaacagc tgctgggatt acacttggca tggacgagct gtacaagtaa
     2041 gaattcgtaa tcatgtcata gctgtttcct gtgtgttaat tcagaacgct cggttgccgc
     2101 cgggcgtttt ttatgcagct cgcgatatat cctgatgtat ttttaatttt gcaccatttc
     2161 caaaaggata agtgacataa gtaaaaaggc atcatcggga gttatcctat caggaaaacc
     2221 aagataatac ctaagtagaa agtgttcaat ccgtgttaaa ttgggaaata tcatccataa
     2281 actttattac tcatactata attcaatttt aacgtcttcg tccatttggg cttcaaattc
     2341 atcgagtagt gctcgtgctt ctgcaattga ttgtgtgttc atcaattgat gtcgaagttc
     2401 gctagcgcct cttatgccac gcacatagat tttaaagaat ctacgcaagc tcttgaattg
     2461 tcgtatttca tctttttcat atttgttaaa caatgataaa tgcaatctca atagatctaa
     2521 tagttccttg cttgtgtgtt cgcgtggttc tttttcaaaa gcgaatggat tgtggaaaat
     2581 gcctctacca atcatgacgc catcaatgcc atatttttct gccagttcaa gtcctgtttt
     2641 tctatcggga atatcaccgt taattgttaa caatgtattt ggtgcaattt cgtcacgtaa
     2701 atttttaata gcttcgatta attcccaatg tgcatctact ttactcattt ctttacgtgt
     2761 acgaagatga atagataaat tggcaatgtc ttgttcgaag acgtgcttca accaatcttt
     2821 ccattcatcg atttcatagt agccaaggcg tgttttaaca cttaccggaa gcccacctgc
     2881 tttagtcgct tgaataattt cggcagcaac gtcaggtctt aagattaagc cggaaccctt
     2941 acggtacccg gggatcctct agagtcgacc tgcaggcatg caagctctga tttcactttt
     3001 tgcattctac aaactgcata actcatatgt aaatcgctcc tttttaggtg gcacaaatgt
     3061 gaggcatttt cgctctttcc ggcgaggcta gttaccctta agttattggt atgactggtt
     3121 ttaagcgcaa aaaaagttgc tttttcgtac ctattaatgt atcgttagaa aaccgactgt
     3181 aaaaagtaca gtcggcatta tctcatatta taaaagccag tcattaggcc tatctgacaa
     3241 ttcctgaata gagttcataa acaatcctgc atgataacca tcacaaacag aatgatgtac
     3301 ctgtaaagat agcggtaaat atattgaatt acctttatta atgaattttc ctgctgtaat
     3361 aatgggtaga aggtaattac tattattatt gatatttaag ttaaacccag taaatgaagt
     3421 ccatggaata atagaaagag aaaaagcatt ttcaggtata ggtgttttgg gaaacaattt
     3481 ccccgaacca ttatatttct ctacatcaga aaggtataaa tcataaaact ctttgaagtc
     3541 attctttaca ggagtccaaa taccagagaa tgttttagat acaccatcaa aaattgtata
     3601 aagtggctct aacttatccc aataacctaa ctctccgtcg ctattgtaac cagttctaaa
     3661 agctgtattt gagtttatca cccttgtcac taagaaaata aatgcagggt aaaatttata
     3721 tccttcttgt tttatgtttc ggtataaaac actaatatca atttctgtgg ttatactaaa
     3781 agtcgtttgt tggttcaaat aatgattaaa tatctctttt ctcttccaat tgtctaaatc
     3841 aattttatta aagttcattt gatatgcctc ctaaattttt atctaaagtg aatttaggag
     3901 gcttacttgt ctgctttctt cattagaatc aatccttttt taaaagtcaa tattactgta
     3961 acataaatat atattttaaa aatatcccac tttatccaat tttcgtttgt tgaactaatg
     4021 ggtgctttag ttgaagaata aagaccacat taaaaaatgt ggtcttttgt gtttttttaa
     4081 aggatttgag cgtagcgaaa aatccttttc tttcttatct tgataataag ggtaactatt
     4141 gccgggatag actgtaacat tctcacgcat aaaatcccct ttcattttct aatgtaaatc
     4201 tattacctta ttattaattc aattcgctca taattaatcc tttttcttat tacgcaaaat
     4261 ggcccgattt aagcacaccc tttattccgt taatgcgcca tgacagccat gataattact
     4321 aatactagga gaagttaata aatacgggtg atctaatgat tcaatgattc aaacccttgt
     4381 gaacttcttt agaacaaaag aggttcgtaa caagattttc ttcacactag caatgttagt
     4441 aatttttaaa atagggactt atataccagc tccaggagta aatcctgcag cttttgataa
     4501 tccccaaggt tctcaaggtg ccactgagtt attaaatact tttggtggcg gagccttgaa
     4561 acgattttct atttttgcaa tgggtattgt accctacatc actgcatcaa tcgtaatgca
     4621 attattacaa atggatattg tccctaaatt ctcagaatgg gcaaaacaag gtgaagtagg
     4681 tagaagaaag ttaaataacg ttactcgtta tttagcaatt tctttagcat ttatccaatc
     4741 tataggtatg gcattccaat ttaataatta tctcaaaggt gcgctgatta tcaatcagtc
     4801 aattatgagt tatttattaa tagcactagt tttgacagca ggaactgctt tcttaatatg
     4861 gcttggtgat caaatcactc agttcggtgt tggtaatggt atttctatta tcatattccc
     4921 atcaagctta ttttaattat actctatcaa tgatagagtg tcaatatttt ttttagtttt
     4981 tcatgaactc gatcgagggg atccaaataa aaaactagtt tgacaaataa ctctatcaat
     5041 gatagagtgt caacaaaaag gaggaattaa tgatgtctag attagataaa agtaaagtga
     5101 ttaacagcgc attagagctg cttaatgagg tcggaatcga aggtttaaca acccgtaaac
     5161 tcgcccagaa gctaggtgta gagcagccta cattgtattg gcatgtaaaa aataagcggg
     5221 ctttgctcga cgccttagcc attgagatgt tagataggca ccatactcac ttttgccctt
     5281 tagaagggga aagctggcaa gattttttac gtaataacgc taaaagtttt agatgtgctt
     5341 tactaagtca tcgcgatgga gcaaaagtac atttaggtac acggcctaca gaaaaacagt
     5401 atgaaactct cgaaaatcaa ttagcctttt tatgccaaca aggtttttca ctagagaatg
     5461 cattatatgc actcagcgct gtggggcatt ttactttagg ttgcgtattg gaagatcaag
     5521 agcatcaagt cgctaaagaa gaaagggaaa cacctactac tgatagtatg ccgccattat
     5581 tacgacaagc tatcgaatta tttgatcacc aaggtgcaga gccagccttc ttattcggcc
     5641 ttgaattgat catatgcgga ttagaaaaac aacttaaatg tgaaagtggg tcttaaaagc
     5701 agcataacct ttttccgtga tggtaacttc acggtaacca agataacaaa gaatacaaga
     5761 aaatatttac aaaaaatcaa tttaacaatt ccttaaaaca tgcaggaatt gacgatttaa
     5821 acaatattag ctttgaacaa ttcttatctc ttttcaatag ctataaatta tttaataagt
     5881 aagttaaggg atgcataaac tgcatccctt aacttgtttt tcgtgtgcct attttttgtg
     5941 aatcgattat gtcttttgcg cagtcggctt aaaccagttt tcgctggtgc gaaaaaagag
     6001 tgtcttgtga cacctaaatt caaaatctat cggtcagatt tataccgatt tgattttata
     6061 tattcttgaa taacatacgc cgagttatca cataaaagcg ggaaccaatc atcaaattta
     6121 aacttcattg cataatccat taaactctta aattctacga ttccttgttc atcaataaac
     6181 tcaatcattt ctttaattaa tttatatcta tctgttgttg ttttctttaa taattcatca
     6241 acatctacac cgccataaac tatcatatct tctttttgat atttaaattt attaggatcg
     6301 tccatgtgaa gcatatatct cacaagacct ttcacacttc ctgcaatctg cggaatagtc
     6361 gcattcaatt cttctgtaat tatttttatc tgttcataag atttattacc ctcatacatc
     6421 actagaatat gataatgctc ttttttcatc ctatcttctg tatcagtatc cctatcatgt
     6481 aatggagaca ctacaaattg aatgtgtaac tcttttaaat actctaacca ctcggctttt
     6541 gctgattctg gatataaaac aaatgtccaa ttacgtcctc ttgaattttt cttgttttca
     6601 gtttctttta ttacattttc gctcatgata taataacggt gctaatacat ttaacaaaat
     6661 ttagtcatag ataggcagca tgccagtgct gtctatcttt ttttgtttaa aatgcaccgt
     6721 attcctcctt tgcatatttt tttattagaa taccggttgc atctgatttg ctaatattat
     6781 atttttcttt gattctattt aatatctcat tttcttctgt tgtaagtctt aaagtaacag
     6841 caactttttt ctcttctttt ctatctacaa ccatcactgt acctcccaac atctgttttt
     6901 ttcactttaa cataaaaaac aaccttttaa cattaaaaac ccaatattta tttatttgtt
     6961 tggacaatgg acaatggaca cctagggggg aggtcgtagt acccccctat gttttctccc
     7021 ctaaataacc ccaaaaatct aagaaaaaaa gacctcaaaa aggtctttaa ttaacatctc
     7081 aaatttcgca tttattccaa tttccttttt gcgtgtgatg cgctgcgtcc attaaaaatc
     7141 ctagagcttt gcaaccgaaa gttaatagct gtcgctacta ctttcgctta cgctctaagt
     7201 atattttaag gactgtcaca cgcaaaaagt tttctcggca taaaagtacc tctacatctc
     7261 taaatcgtct gtacgctgtt tctcacgctt tctatcgacc ttctggacat tatcctgtac
     7321 aacatccata aactgtccca cacgctcgaa tttggaatca ttaaagaatt tctctttaag
     7381 cctattaaac cctttctcaa acccagggaa attcgccctc gcagcacgat ataaagtcac
     7441 tgtactatct tgaaatttct ctgatacatt caactgctca ttcaaactat cattctctcg
     7501 ctttaattta ttaacctctt tacttttttc gtgatacccc tctttccatg tattcactac
     7561 ttctttcaaa ctctctctac gtttttttaa ttcttgattt tctgtgtaat agtctgtgct
     7621 cttaatattt tcgtaatcat caacaatccg ttcctgcctc gcgcgtttcg gtgatgacgg
     7681 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     7741 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     7801 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
     7861 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     7921 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     7981 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     8041 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     8101 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     8161 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     8221 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     8281 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     8341 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     8401 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     8461 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     8521 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     8581 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     8641 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     8701 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     8761 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     8821 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     8881 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     8941 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     9001 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     9061 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     9121 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     9181 gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     9241 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     9301 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     9361 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     9421 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     9481 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     9541 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     9601 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     9661 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     9721 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     9781 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     9841 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     9901 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaa
//
