LOCUS       sequence-209915-        8096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A nourseothricin resistance marker used to construct gene 
            disruptions in Candida glabrata. The natMX6 marker can be evicted by
            growth on media lacking methionine and containing estradiol..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8096)
  AUTHORS   Vu BG, Moye-Rowley WS
  TITLE     Construction and Use of a Recyclable Marker To Examine the Role of 
            Major Facilitator Superfamily Protein Members in Candida glabrata 
            Drug Resistance Phenotypes.
  JOURNAL   mSphere. 2018 Mar 28;3(2). pii: mSphere00099-18. doi: 
            10.1128/mSphere.00099-18. eCollection 2018 Mar-Apr.
  PUBMED    29600281
REFERENCE   2  (bases 1 to 8096)
  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: "mSphere."; 
            date: "2018-03-28"; volume: "3(2). pii: mSphere00099-18. doi"; 
            pages: " 10.1128/mSphere.00099-18. eCollection 2018 Mar-Apr"
COMMENT     Sequence Label: pBV65
FEATURES             Location/Qualifiers
     source          1..8096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    53..86
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        141..484
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      1063..1260
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             4262..5197
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAVNLWPSPLMIKRSKENSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLVTSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLLLSHIRHMSNEGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     terminator      5374..5621
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5390..5408)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    5647..5680
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(5733..5752)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5734..5752)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5839..5856)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6010..6598)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6090..6109)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6769..7560,7561..7629))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7392..7411
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7630..7734)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7802..7820
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7858..7880)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7980..7999
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(8080..8096,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(8080..8096,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acaaccctta atataacttc
       61 gtataatgta tgctatacga agttattagg tctagagatc tgtttagctt gcctcgtccc
      121 cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac agtcttgacg
      181 tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata catccccatg
      241 tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa aattacggct
      301 cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg aattgtcccc
      361 acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg ttcttctttc
      421 atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc gaacataaac
      481 aaccatgggt acaaccttag acgatacagc atacagatac agaacaagtg tccctggtga
      541 cgctgaagcc attgaagcct tagacggtag ttttactaca gacactgtat tcagagttac
      601 cgcaactggt gatggtttta ctttgagaga agttccagtc gatccacctt taacaaaagt
      661 tttccctgat gacgaatccg atgacgaaag tgatgacggt gaagatggtg acccagattc
      721 aagaacattt gttgcttatg gtgatgacgg tgatttggcc ggtttcgttg tcgtatctta
      781 ctcaggttgg aacagaagat taaccgttga agatatagaa gtcgctccag aacatagagg
      841 tcacggtgtt ggtagagcat tgatgggttt agccaccgaa tttgctagag aaagaggtgc
      901 tggtcatttg tggttagaag tcactaatgt aaacgctcct gcaatacacg cctatagaag
      961 aatgggtttc acattgtgtg gtttagatac cgctttgtac gacggtacag cctccgacgg
     1021 tgaacaagcc ttatacatgt ctatgccatg cccttaatca gtactgacaa taaaaagatt
     1081 cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa
     1141 tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag
     1201 ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg
     1261 ctgtcgattc gatactaacg ccgccatcca gttttttata ttatttggtc taacatacca
     1321 gttacaatta gtattacaat ggtttaccaa gtacaaatta tgtaatatgg ttataattgg
     1381 cttatatgta tggaggtagg gaactgcagg ttattgttct gttaaactac cagtaatgca
     1441 agacggctaa gtcatatgac tgccactttg tcaatcctga agaaaatgac aatagaaagt
     1501 gatataatac acgtgatatg cgtaggatgg gacccaaaga tagccacatc acatgatctt
     1561 caaaaacccc gtcaaaaagg aataaatcga aggaaaaaaa atgccacgtg actttgatag
     1621 tcaaaatcag gttatattac tatacgatcc cccacacata acctttacca cacaccgcac
     1681 gggctggact tatcctatta gaaagcggtg cagccagggc caagaaaacg cgaacgacgc
     1741 agaaaaaacg tcaagcaaaa aaactgtggt gttatttaaa gcatacaatt tcctgcctcc
     1801 ctttaatgct acgggggatc tagcaaatgg gaaaatatca catgactttg gctagaaggg
     1861 gtgggaaaac agatatttta gtcacattgt ttgatttcac gtactacacg atacactaca
     1921 ctacacaata cactgcaata cactacactg tatggttccc tccggccctg gatatgctag
     1981 cgaaggatcc caagcccatc gaggaaatca ttcaagagca tctgcaggtc tcaaatacac
     2041 taagtccaaa cacaaacagc actgagctaa tccaagacac caattgcatg ccttctcaat
     2101 tagaacagtt tattacacag cttataactg tgcatcttgc cattcttttc gagatagcca
     2161 tttgcatttg attaacatgc ttagctcgtt ctcaagccac agtaaaatgg attgcctttt
     2221 gagtttccat cgttgtataa ataggcactc attcctatag ccttgctctt tggtcttgtc
     2281 tctgtaatgg taatagctgt aagtcaggat ataacactcc agaaaagaaa cacctaacaa
     2341 tatgtccaat ttactgaccg gtatgttcga ctactatgga tagacaagac tataactcct
     2401 ataatatagg gcgtgtggtc gctgataaac aatttcttct acacacaaca aggagttacc
     2461 cagtgttgac tagagtatgg ccataatcta gtcaaatact ctagagatga ctctcgctat
     2521 ttcattcaat tgcattaata ctaacaattt ctattttaaa attttaaact actttctact
     2581 gcattgaact tcattttggt tttttctacg gtttttcttt tgatctcctt ttcttatctg
     2641 tccctgcgtg tgtgaataac cgctgcgata ctcaattgct ttcataacca ttttcacagg
     2701 atcaattgct gagactatcc tctcacctct ctatcgtttc cccctttgcc acattagcct
     2761 atggcacgta catttttttt tcaccctgtg cgaagcgaca attttttttt tgggaaaatc
     2821 atgataggcg cgtgcgtggg tttctaaggg tgacggcgat tatgagttag gagtggtggg
     2881 cattgggatc atacatataa tgtaaaagtg gcaacggttg atgaacaggc acaactacct
     2941 gcacagatgc tactatggcc tagggctttg cgtttctcta cttattgaga tgcttttgtt
     3001 ttaggtgaga aatcgaaagc catggctaca aaataggatc tacctctaca acatatgtag
     3061 tccacttcca tcagttattg gccggtaatg gtgttcacac attgcaatta ggtaaatgaa
     3121 agtcttcccc cagcagaagg taagatcaac attaggagaa gaccaaagta aagaagacca
     3181 aatgattata gttcagggat aaagaagtgg ttttttttta aaaaccagta tatgtacagt
     3241 acaccaaaat ttgcctgcat taccggtcga tgcaacgagt gatgaggttc gcaagaacct
     3301 gatggacatg ttcagggatc gccaggcgtt ttctgagcat acctggaaaa tgcttctgtc
     3361 cgtttgccgg tcgtgggcgg catggtgcaa gttgaataac cggaaatggt ttcccgcaga
     3421 acctgaagat gttcgcgatt atcttctata tcttcaggcg cgcggtctgg cagtaaaaac
     3481 tatccagcaa catttgggcc agctaaacat gcttcatcgt cggtccgggc tgccacgacc
     3541 aagtgacagc aatgctgttt cactggttat gcggcggatc cgaaaagaaa acgttgatgc
     3601 cggtgaacgt gcaaaacagg ctctagcgtt cgaacgcact gatttcgacc aggttcgttc
     3661 actcatggaa aatagcgatc gctgccagga tatacgtaat ctggcatttc tggggattgc
     3721 ttataacacc ctgttacgta tagccgaaat tgccaggatc agggttaaag atatctcacg
     3781 tactgacggt gggagaatgt taatccatat tggcagaacg aaaacgctgg ttagcaccgc
     3841 aggtgtagag aaggcactta gcctgggggt aactaaactg gtcgagcgat ggatttccgt
     3901 ctctggtgta gctgatgatc cgaataacta cctgttttgc cgggtcagaa aaaatggtgt
     3961 tgccgcgcca tctgccacca gccagctatc aactcgcgcc ctggaaggga tttttgaagc
     4021 agctcatcga ctgatttacg gcgctgagga tgactctggt cagaggtacc tggcctggtc
     4081 tggacacagt gcccgtgtcg gagccgcgcg agatatggcc cgcgctggag tttcaatacc
     4141 ggagatcatg caagctggtg gctggaccaa tgtaaatatt gtcatgaact atatccgtaa
     4201 cctggatagt gaaacagggg caatggtggt cctgctggaa gatggcgatc tcgagccatc
     4261 cgctggagac atgagagctg tcaacctttg gccaagcccg ctcatgatca aacgctctaa
     4321 ggagaacagc ctggccttgt ccctgacggc cgaccagatg gtcagtgcct tgttggatgc
     4381 tgagcccccc atactctatt ccgagtatga tcctaccaga cccttcagtg aagcttcgat
     4441 gatgggctta ctgaccaacc tggcagacag ggagctggtt cacatgatca actgggcgaa
     4501 gagggtgcca ggctttgtgg atttgaccct ccatgatcag gtccaccttc tagaatgtgc
     4561 ctggctagag atcctgatga ttggtctcgt ctggcgctcc atggaacacc cggggaagct
     4621 cctgtttgct cctaacttgc tcctggacag gaatcaaggt aaatgtgtgg aaggcatggt
     4681 ggagatcttc gacatgctgc tggttacatc atctcggttc cgcatgatga atctgcaggg
     4741 agaggagttt gtgtgcctca aatctattat tttgcttaat tctggagtat acacatttct
     4801 gtccagcacc ctgaagtctc tcgaggagaa ggaccatatc caccgagtcc tggacaagat
     4861 cacagacact ttgatccacc tgatggccaa ggcaggcctg accctgcagc agcagcacca
     4921 gcggctagcc cagctcctcc tcctcctctc ccacatcagg cacatgagta acgaaggcat
     4981 ggagcatctg tacagcatga agtgcaagaa cgtggtaccc ctctatgacc tgctgctgga
     5041 gatgctagac gcccaccgcc tacatgcgcc cactagtcgt ggaggggcat ccgtggagga
     5101 gacggaccaa agccacttgg ccactgcggg ctctacttca tcgcattcct tgcaaaagta
     5161 ttacatcacg ggggaggcag agggtttccc tgccacagtc tgagtagaat gcctgatatt
     5221 gactcaatat ccgttgcgtt tcctgtcaaa agtatgcata gtgctgaaca tttcgtgatg
     5281 aatgccaccg aggaagaagc acggcgcggt tttgctaaag tgatgtctga gtttggcgaa
     5341 ctcttgggta aggttggaat tgtcgacctc gagtcatgta attagttatg tcacgcttac
     5401 attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag
     5461 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     5521 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     5581 tgcttgagaa ggttttggga cgctcgaagg ctttaatttg caaacgagct ctcgagaacc
     5641 cttaatataa cttcgtataa tgtatgctat acgaagttat taggtgatat cagatccact
     5701 agtggcctat gcggccgcgg atctgccggt ctccctatag tgagtcgtat taatttcgat
     5761 aagccaggtt aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5821 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5881 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5941 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6001 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6061 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6121 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6181 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6241 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6301 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6361 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6421 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6481 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6541 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6601 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6661 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6721 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6781 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6841 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6901 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6961 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7021 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7081 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7141 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7201 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7261 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7321 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7381 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7441 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7501 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7561 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7621 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7681 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7741 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     7801 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     7861 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     7921 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     7981 ggcatcagag cagattgtac tgagagtgca ccatatggac atattgtcgt tagaacgcgg
     8041 ctacaattaa tacataacct tatgtatcat acacatacga tttaggtgac actata
//
