LOCUS       sequence-368242-        8248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of reverse transactivator (CgrtTA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8248)
  AUTHORS   Schrevens S, Sanglard D
  TITLE     A novel Candida glabrata doxycycline-inducible system for in 
            vitro/in vivo use.
  JOURNAL   FEMS Yeast Res. 2022 Sep 24;22(1):foac046. doi: 
            10.1093/femsyr/foac046.
  PUBMED    36047937
REFERENCE   2  (bases 1 to 8248)
  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: "FEMS Yeast 
            Res."; date: "2022-09-24"; volume: "22(1):foac046. doi"; pages: " 
            10.1093/femsyr/foac046"
COMMENT     Sequence Label: pSSa26
FEATURES             Location/Qualifiers
     source          1..8248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..38)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(18..36)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(57..73)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(57..73)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(70..92)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    81..97
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(105..111,112..129,130..135))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    150..171
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(288..305)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(459..1047)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(539..558)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1218..2009,2010..2078))
                     /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     1841..1860
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2079..2150)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2788..2806
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2844..2866)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2966..2985
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3009..3224
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             complement(3841..4869)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             complement(3841..4869)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     rep_origin      complement(5572..6027)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5709..5730)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5921..5940
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6153..6175
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6167..6184
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6168..6184
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6194..6213
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6194..6212
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(7001..7021)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
       61 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
      121 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
      181 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
      241 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
      301 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
      361 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
      421 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
      481 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      541 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
      601 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
      661 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
      721 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      781 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
      841 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
      901 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
      961 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1021 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1081 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     1141 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     1201 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     1261 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     1321 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     1381 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     1441 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     1501 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     1561 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     1621 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     1681 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     1741 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     1801 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     1861 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     1921 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     1981 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     2041 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     2101 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tggggatctc
     2161 taatgatgca atttttgttt tgtacactaa gattctattt cggaactctt acccatgtca
     2221 ttatttactt tattttcatt attgtaattt tttaagcttt tattatgaat tatttaaata
     2281 caattctttt tgatcatgtg acagacatca ctgaaagatg tgctctgcgt tattagtcag
     2341 attgaaatat acattttcat gtagatatga tgttcttact ttaaatgttg ttccatttgc
     2401 tagcttaaat agacgatatt ataaatttaa gtattcacta aaaatctgac ccattaatgt
     2461 tagaattagt attatttttt gtgatattac atttttatat tttggatttc aaccaatgct
     2521 ctgcgatatg attcaacaga ataatatgta caagctactt taattcatat atactaatag
     2581 attcacagat ccagtacgac tgcaagaatt tacgttttgt ttgaaatggt agtagctaca
     2641 ctaaattaac aatgatcgat taatccacat ccacagtgtg taatcatcct aatgaaaaca
     2701 agaataacta caaaaatgaa tataaacttt ttgcatgtta gtctaacctg ctaagaaacc
     2761 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     2821 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     2881 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     2941 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     3001 accacctttt caattcatca tttttttttt attctttttt ttgatttcgg tttccttgaa
     3061 atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta
     3121 gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa
     3181 cccaactgca cagaacaaaa acctgcagga aacgaagata aatcatgtcg aaagctacat
     3241 ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc
     3301 acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg
     3361 agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcgaat
     3421 tcctgcagcc cctttacctc tatatcgtgt tcgtgcgtat ttcttcctct ataatatgaa
     3481 aagaaaggca gtagtcttga ttttctattc ttccaaatat tacaagaata cccagaacca
     3541 caagaagcag aagaaactaa ctgctttata tacttaattt cccgccacaa gctcatcgcc
     3601 cagaagtttt tttcagtttt tcacctgaca atctggcagc tcgctgaaaa attttcacct
     3661 ggggttgcct gctacgtaaa gtgatcacgt ttatgaatgg cacattatat agtggcccag
     3721 acggtctgat ggtagttcat gtagttaaga ataaatactc tgtttgtgtg atttctatgt
     3781 gtacgttata tatataaata ttgttgatgg tggtagctgt gggttgtgtt cgggggatct
     3841 ttattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg cgagtacttc
     3901 tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac gcccgacagt
     3961 cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa
     4021 attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca tatacgcccg
     4081 gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg tctgctgctc
     4141 catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt gggaatcccc
     4201 gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg tcaggacatt
     4261 gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct cggcccaaag
     4321 catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg
     4381 ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg
     4441 accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat cggccgcagc
     4501 gatcgcatcc atggcctccg cgaccggctg cagaacagcg ggcagttcgg tttcaggcag
     4561 gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa
     4621 ttccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac
     4681 ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat atccacgccc
     4741 tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca tcaggtcgga
     4801 gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga gttcaggctt
     4861 tttacccata gatccactag atgttttata tttgttgtaa aaagtagata attacttcct
     4921 tgatgatctg taaaaaagag aaaaagaaag catctaagaa cttgaaaaac tacgaattag
     4981 aaaagaccaa atatgtattt cttgcattga ccaatttatg caagtttata tatatgtaaa
     5041 tgtaagtttc acgaggttct actaaactaa accaccccct tggttagaag aaaagagtgt
     5101 gtgagaacag gctgttgttg tcacacgatt cggacaattc tgtttgaaag ggagagagta
     5161 acagtacgat cgaacgaact ttgctctgga gatcacagtg ggcatcatag catgtggtac
     5221 taaacccttt cccgccattc cagaaccttc gattgcttgt tacaaaacct gtgagccgtc
     5281 gctaggacct tgttgtgtga cgaaattgga agctgcaatc aataggaaga caggaagtcg
     5341 agcgtgtctg ggttttttca gttttgttct ttttgcaaac aaatcacgag cgacggtaat
     5401 ttctttctcg ataagaggcc acgtgcttta tgggggggat cgaagttcct attctctaga
     5461 atgcatgtat actaaactca caaattagag cttcaattta attatatcag ttattaccct
     5521 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa
     5581 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     5641 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     5701 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     5761 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     5821 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     5881 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     5941 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     6001 cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga
     6061 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     6121 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     6181 cagtgagcgc gcgtaatacg actcactata gggcgaattg ggtaccttta cctctatatc
     6241 gtgttcgtgc gtatttcttc ctctataata tgaaaagaaa ggcagtagtc ttgattttct
     6301 attcttccaa atattacaag aatacccaga accacaagaa gcagaagaaa ctaactgctt
     6361 tatatactta atttcccgcc acaagctcat cgcccagaag tttttttcag tttttcacct
     6421 gacaatctgg cagctcgctg aaaaattttc acctggggtt gcctgctacg taaagtgatc
     6481 acgtttatga atggcacatt atatagtggc ccagacggtc tgatggtagt tcatgtagtt
     6541 aagaataaat actctgtttg tgtgatttct atgtgtacgt tatatatata aatattgttg
     6601 atggtggtag ctgtgggttg tgttctcgac cgggggatcc ttattccttc tttggatttg
     6661 gtggagtatc ttcatcatcg aacaagtaat tataaacatc atccatagta gtagtattaa
     6721 acataccagt agtaataccg aaagcattat aagcagtttg atcagtccaa cctggagaca
     6781 atggcttaga attgttacca tcatcaatct tagaagcagt aatttcgttg ttcataaaat
     6841 tgtcatgaac attagaagaa gcaattggtt gagacaaagc ttgagagtta gtttgagtag
     6901 tgatcaagtt tggaccatta gaagagttag tgaaagtgaa agacaaagaa gagtcagcga
     6961 tgttaccaga ttgattaaaa ttagcagctg tacccaattc aacctttctc ttcttttttg
     7021 gaggctctgg gatcaactca gccttatcag aagcttccaa agaaccagat tcgcatttca
     7081 attgcttctc caaaccacag ataatcaatt ccaaaccaaa caaaaaagct ggctctgcac
     7141 cctgtctgtc gaacaattct atagcctgtt tcaacaatgg aggcatagag tctgttgtag
     7201 gtgtttctct ctcttctttt gcaacctgat gctcttgctc ttccaaaacg caacccaaag
     7261 taaaatgacc aacagcagac aaagcataca atgcattttc caaagagaaa ccctgttgac
     7321 acaaaaaagc caattgattt tccaatgttt catactgttt ttctgtaggt cttgtaccca
     7381 aatgaacctt agcaccatct ctgtgagaca acaatgcaca tctgtaagac ttagcgttgt
     7441 ttctcaaaaa gtcttgccaa gactcaggct ccaatggaca agagtgtgtg tgatgtctat
     7501 ccaacatttc tattggcaat gcatccaaca atgctctttt atttttaacg tgccagtaca
     7561 atgtaggctg ctcaacaccc aatttttgtg ccaattttct agtagtcaaa ccttctatac
     7621 caacaccgtt caacaactcc aaagcaccat taattatttt agatttgtcc aatctagaca
     7681 ttttactagt tctagatgtt ttatatttgt tgtaaaaagt agataattac ttccttgatg
     7741 atctgtaaaa aagagaaaaa gaaagcatct aagaacttga aaaactacga attagaaaag
     7801 accaaatatg tatttcttgc attgaccaat ttatgcaagt ttatatatat gtaaatgtaa
     7861 gtttcacgag gttctactaa actaaaccac ccccttggtt agaagaaaag agtgtgtgag
     7921 aacaggctgt tgttgtcaca cgattcggac aattctgttt gaaagggaga gagtaacagt
     7981 acgatcgaac gaactttgct ctggagatca cagtgggcat catagcatgt ggtactaaac
     8041 cctttcccgc cattccagaa ccttcgattg cttgttacaa aacctgtgag ccgtcgctag
     8101 gaccttgttg tgtgacgaaa ttggaagctg caatcaatag gaagacagga agtcgagcgt
     8161 gtctgggttt tttcagtttt gttctttttg caaacaaatc acgagcgacg gtaatttctt
     8221 tctcgataag aggccacgtg ctttatgg
//
