LOCUS       sequence-403443-        6260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ERG11-3X Minidegron-5X FLAG-HIS3MX6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6260)
  TITLE     Plasmids for auxin-inducible degron tagging of genes in Candida 
            glabrata
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6260)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pBV58
FEATURES             Location/Qualifiers
     source          1..6260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1101..1118)
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
     CDS             1124..1327
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     CDS             1346..1549
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     CDS             1574..1777
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     CDS             1787..1852
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1856..1879
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1883..1906
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      1942..2129
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1997..2021)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2204..2547
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2548..3246
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      3250..3447
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(4039..4055)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4039..4055)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4052..4074)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4063..4079
                     /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(4087..4093,4094..4111,4112..4117))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4132..4153
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4270..4287)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4441..5029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4521..4540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5200..5991,5992..6060))
                     /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     5823..5842
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6061..6165)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6233..6251
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt accatttggc
      421 tgaaagacca gatgtccaag aagaattata ccaagaacaa atgcgcgtct tgaacaacga
      481 taccaaggaa ttgacttacg atgacctaca aaacatgcct ctattgaacc aaatgatcaa
      541 ggaaactttg agattgcacc acccattgca ctctttgttc cgtaaagtca tgagagatgt
      601 cgctattcca aacacttcct acgttgtccc aagggactac cacgttctag tctccccagg
      661 ttacactcac ttgcaagaag aattcttccc taagccaaat gaattcaaca tccaccgttg
      721 ggacggtgat gctgcttctt ccagtgctgc tggtggtgac gaagttgatt acggtttcgg
      781 tgctatctcc aagggtgttt cctctccata cttgccattc ggtggtggta gacacagatg
      841 tatcggtgaa ttgttcgctt actgtcaatt gggtgtgttg atgtccattt tcatcagaac
      901 catgaaatgg cgttacccaa ctgaaggtga aactgtccca ccatctgact tcacctccat
      961 ggtcacccta ccaactgccc ctgctaagat ctactgggaa aagagacatc aagaacaaaa
     1021 gtaccactat agaacgcggc cgccagctga agcttcgtac gctgcaggtc gacggatccc
     1081 cgggttaatt aatgccggtg ctggtgctgg tgctggtgcc ggtaaagaaa agagtgcttg
     1141 cccaaaagac cctgctaaac ctcctgccaa ggctcaagtt gtaggatggc cacctgttag
     1201 atcatacaga aagaatgtca tggtttcttg tcaaaagtct tcaggtggac cagaagctgc
     1261 agcctttgtt aaagtatcta tggatggtgc cccatatttg agaaagattg acttaaggat
     1321 gtacaagggt gctggagcag gcgccaaaga aaaatctgcc tgtccaaaag atcctgcaaa
     1381 gccacctgct aaagcacaag tcgtgggttg gcctcctgtt agaagttata gaaagaatgt
     1441 gatggtttct tgccaaaaat ctagtggcgg tcctgaagcc gcagccttcg tcaaagtatc
     1501 tatggacggc gctccttatt tgcgtaagat agacttgcgt atgtataaag gtgccggagc
     1561 tggcgcaggt gctaaggaaa aatctgcgtg tccaaaggac cctgcaaaac caccagccaa
     1621 ggcacaagtt gtaggttggc cccctgtaag atcctataga aagaatgtta tggtttcttg
     1681 ccaaaaatct tctggaggcc ctgaagcagc tgcatttgtt aaagttagta tggacggtgc
     1741 tccttacttg agaaaaatag acttgagaat gtataaaaga tctatggatt ataaagatca
     1801 tgatggtgac tacaaggacc acgacattga ttacaaggat gatgacgaca aaatggacta
     1861 caaggacgac gacgacaagt tggattataa agatgatgat gataaataag ctcagtgctg
     1921 aggcgcgcca cttctaaata agcgaatttc ttatgattta tgatttttat tattaaataa
     1981 gttataaaaa aaataagtgt atacaaattt taaagtgact cttaggtttt aaaacgaaaa
     2041 ttcttattct tgagtaactc tttcctgtag gtcaggttgc tttctcaggt atagtatgag
     2101 gtcgctctta ttgaccacac ctctaccggc agatccgcta gggataacag ggtaatatag
     2161 atctgtttag cttgcctcgt ccccgccggg tcacccggcc agcgacatgg aggcccagaa
     2221 taccctcctt gacagtcttg acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta
     2281 catttagccc atacatcccc atgtataatc atttgcatcc atacattttg atggccgcac
     2341 ggcgcgaagc aaaaattacg gctcctcgct gcagacctgc gagcagggaa acgctcccct
     2401 cacagacgcg ttgaattgtc cccacgccgc gcccctgtag agaaatataa aaggttagga
     2461 tttgccactg aggttcttct ttcatatact tccttttaaa atcttgctag gatacagttc
     2521 tcacatcaca tccgaacata aacaaccatg gcagaaccag cccaaaaaaa gcaaaaacaa
     2581 actgttcagg agcgcaaggc gtttatctcc cgtatcacta atgaaactaa aattcaaatc
     2641 gctatttcgc tgaatggtgg ttatattcaa ataaaagatt cgattcttcc tgcaaagaag
     2701 gatgacgatg tagcttccca agctactcag tcacaggtca tcgatattca cacaggtgtt
     2761 ggctttttgg atcatatgat ccatgcgttg gcaaaacact ctggttggtc tcttattgtt
     2821 gaatgtattg gtgacctgca cattgacgat caccatacta ccgaagattg cggtatcgca
     2881 ttagggcaag cgttcaaaga agcaatgggt gctgtccgtg gtgtaaaaag attcggtact
     2941 gggttcgcac cattggatga ggcgctatca cgtgccgtag tcgatttatc tagtagacca
     3001 tttgctgtaa tcgaccttgg attgaagaga gagatgattg gtgatttatc cactgaaatg
     3061 attccacact ttttggaaag tttcgcggag gcggccagaa ttactttgca tgttgattgt
     3121 ctgagaggtt tcaacgatca ccacagaagt gagagtgcgt tcaaggcttt ggctgttgcc
     3181 ataagagaag ctatttctag caatggcacc aatgacgttc cctcaaccaa aggtgttttg
     3241 atgtgaagta ctgacaataa aaagattctt gttttcaaga acttgtcatt tgtatagttt
     3301 ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat tttttttcgc
     3361 ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat
     3421 cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg ccatccagtt
     3481 taaacacatt ttcttccaat aatcaatggt caaaacctac ttcaaacaat attgctgtct
     3541 ggctcgtata cgggatatac gctgattcat tagcctaata gtaagtcatc gtgaatatca
     3601 acatggatat ttcggcatga cttaagctgg ttgtttcgtt tagagggctt tgctgctaaa
     3661 cgaactatac gatgtcttca tccgtataac tagcaatgta tgcagtgcaa gcatatgaca
     3721 aagttggttg atcccaaaga ttgttttaaa aaaattcatt ttcatgatca aaattcaatg
     3781 cttcttatac tacatttttt tcattttcat tcatctttat tttggtgcca ttaacatata
     3841 gacttaagtt tcatttagct atttatattt attataaaat ttaaaccgtg tatacaatac
     3901 aatattattt aacaaatcga actcttgtat tattatcaaa ctttaccttg gtaagcctaa
     3961 agaaacacgt tttttgcgcc catgttgttt atatttccaa tgctgtccct gcaggcatgc
     4021 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4081 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4141 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4201 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4261 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4321 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4381 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4441 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4501 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4561 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4621 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4681 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4741 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4801 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4861 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4921 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4981 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5041 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5101 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5161 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5221 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5281 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5341 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5401 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5461 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5521 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5581 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5641 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5701 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5761 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5821 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5881 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5941 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6001 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6061 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6121 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6181 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     6241 tatcacgagg ccctttcgtc
//
