LOCUS       sequence-402940-        7361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminal tagging of target proteins with 3xV5-IAA17 degron.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7361)
  AUTHORS   Milholland KL, Gregor JB, Hoda S, Piriz-Antunez S, Duenas-Santero E,
            Vu BG, Patel KP, Moye-Rowley WS, Vazquez de Aldana CR, Correa-Bordes
            J, Briggs SD, Hall MC
  TITLE     Rapid, efficient auxin-inducible protein degradation in Candida 
            pathogens.
  JOURNAL   mSphere. 2023 Aug 18:e0028323. doi: 10.1128/msphere.00283-23.
  PUBMED    37594261
REFERENCE   2  (bases 1 to 7361)
  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. 
            2023 Aug 18:e0028323. doi: 10.1128/msphere.00283-23."
COMMENT     Sequence Label: pHLP701
FEATURES             Location/Qualifiers
     source          1..7361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(53..75)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     175..194
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        218..433
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             434..1237
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1368..1823)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1505..1526)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1717..1736
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1949..1971
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1963..1980
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1964..1980
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1987..2006
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1987..2005
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2035..2051
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2035..2051
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2089..2130
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2173..2856
                     /codon_start=1
                     /gene="Arabidopsis thaliana AXR3"
                     /product="auxin-responsive IAA17 protein"
                     /label=auxin-inducible degron
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="MGSVELNLRETELCLGLPGGDTVAPVTGNKRGFSETVDLKLNLNN
                     EPANKEGSTTHDVVTFDSKEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSG
                     GPEAAAFVKVSMDGAPYLRKIDLRMYKSYDELSNALSNMFSSFTMGKHGGEEGMIDFMN
                     ERKLMDLVNSWDYVPSYEDKDGDWMLVGDVPWPMFVDTCKRLRLMKGSDAIGLAPRAME
                     KCKSRA"
     terminator      2873..3060
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2928..2952)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(5056..5072)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5080..5096)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5125..5143)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5164..5180)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5164..5180)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5177..5199)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5188..5204
                     /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(5212..5218,5219..5236,5237..5242))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5257..5278
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5395..5412)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5566..6154)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5646..5665)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6325..7116,7117..7185))
                     /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     6948..6967
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7186..7290)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(7358..7361,1..15)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
       61 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
      121 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
      181 cagagcagat tgtactgaga gtgcaccata ccaccttttc aattcatcat ttttttttta
      241 ttcttttttt tgatttcggt ttccttgaaa tttttttgat tcggtaatct ccgaacagaa
      301 ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga
      361 agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa cctgcaggaa
      421 acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc
      481 tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt
      541 ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg
      601 tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc
      661 gctaaaggca ttatccgcca agtacaattt tttactcttc gaagacagaa aatttgctga
      721 cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc
      781 agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc
      841 ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa
      901 gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa
      961 agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga
     1021 ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagacgcat tgggtcaaca
     1081 gtatagaacc gtggatgatg tggtctctac aggatctgac attattattg ttggaagagg
     1141 actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg
     1201 ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc
     1261 atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat taccctatgc
     1321 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt
     1381 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     1441 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     1501 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     1561 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     1621 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     1681 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     1741 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     1801 taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg
     1861 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     1921 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     1981 gaattgtaat acgactcact atagggcgaa ttggagctcc accgcggtgg ccgctctaga
     2041 actagtggat ccggtaaacc aattcctaac cctttacttg gtttggacgg taagccaatc
     2101 cctaaccctt tattggggtt ggattccact cctattccta atccattgtt aggtttggat
     2161 tctaccgatc ctatgggatc tgtggaatta aaccttcgtg agactgaatt atgcttaggt
     2221 ttgcctggtg gggatactgt ggccccagtt acaggtaata aacgtggttt ctccgaaacc
     2281 gtggatttaa agttgaacct taataacgag cctgctaaca aggagggaag tacaacacat
     2341 gatgtcgtta cttttgattc caaagaaaaa tccgcttgtc ctaaggaccc tgcaaaacca
     2401 cctgctaagg cacaggtggt gggatggcca cctgttcgtt catatcgaaa gaacgtgatg
     2461 gtatcttgcc agaagagttc cggtgggcca gaagctgccg ctttcgttaa ggtcagtatg
     2521 gatggtgccc catatcttcg taaaattgac cttagaatgt ataaatcata tgacgaatta
     2581 tctaatgcct tgagtaatat gttttcatct ttcactatgg gaaagcacgg aggggaagag
     2641 gggatgatcg attttatgaa tgagagaaag cttatggact tggtcaacag ttgggactac
     2701 gtcccttctt acgaggacaa agacggagac tggatgttgg tgggtgatgt gccttggcca
     2761 atgttcgttg atacatgcaa gcgtcttaga ttaatgaaag gatcagatgc tattggtctt
     2821 gctcctagag caatggagaa gtgtaagagt agagccgcag atccttaacg aggcgaattt
     2881 cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt
     2941 ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta
     3001 ggtcaggttg ctttctcagg tatagcatga ggtcgctctt attgaccaca cctctaccgg
     3061 catgccgagc aaatgcctgc aaatcgctcc ccatttcacc caattgtaga tatgctaact
     3121 ccagcaatga gttgatgaat ctcggtgtgt attttatgtc ctcagaggac aacacctgtt
     3181 gtaatcgtgt cgagcgtcaa aactagagaa taataaagaa aacgatcttt tcaaaaagaa
     3241 aaaacctttt agttttcctt tgttgttgtt gtgggtgtgt gctatttata ttatatagtt
     3301 tactcataat accataaaat attcggtttg attaggttat tttaataagc taatttgttt
     3361 ctaatcgtgt aatttatgct gtgtatatta agtagtgtgt gcactgccca aaaatgtttg
     3421 ttgtttatag tcggttaaag agaaaaaaga aaaaaagatc catacacaca cgttaattag
     3481 ttgttcaacg taatacactc atattttgtt cttatttgct ttcggtcgct gttctcacca
     3541 agatttattg ccaacgaaac aatttttttt tatatatttt cagatttttc tttttttcct
     3601 ttcctttcct tttctaattt tcactcctgg ttttctttct ttcttagaaa cattatctcg
     3661 atattaatat taaaaaaata taatcattca aaatggacgg tggtatgttt tagtttagct
     3721 tcaattctaa ttgattgatt aatcagttga ttggtttcaa tatgacaaat gggtagggtg
     3781 ggaaaacttc attttcaatt cagatcaaac ttttttgttg tcgacataat atttctcgtt
     3841 tgggatgtta ctgtcacatt aataatacac acacatcagc ttataatttt gaaagtaatt
     3901 tatcagatat gttgtgacga tcagtggaaa tggctaactt caatgtatct gttcttcccc
     3961 tttttcaaag ttcacgtttt ttgattgatt gattgatctg tcggcagtgg tttcaaaacc
     4021 attcggtgag taatcctatc aatcaatgtt acgacaaaag gctcaatatt caaaattgca
     4081 atgttttatg ttttcctacg tgtacttgtg caaggcaatt gattcaacat tgcttttggt
     4141 gtttgacgag tttctagttt ggacttgtgt tgttatctgg gctatacaga tttcccggct
     4201 cactatgaat tttttttttc gacgctcagt gcacacaact ataaacaaca caaacacaaa
     4261 cacagcaaga aaaaaaaaaa acgaacattg aattgaaacc aagccaactg aaaaattcct
     4321 tatttaaatg actgtcatac taacccattt ttatagaaga agttgctgct ttagttatcg
     4381 ataacggttc tcatatgaaa atttcggtga tccctgagca ggtggcggaa acattggatg
     4441 ctgagaacca tttcattgtt cgtgaagtgt tcgatgtgca cctatccgac caaggctttg
     4501 aactatctac cagaagtgtg agcccctacc ggaaggatta catctcggat gatgactctg
     4561 atgaagactc tgcttgctat ggcgcattca tcgaccaaga gcttgtcggg aagattgaac
     4621 tcaactcaac atggaacgat ctagcctcta tcgaacacat tgttgtgtcg cacacgcacc
     4681 gaggcaaagg agtcgcgcac agtctcatcg aatttgcgaa aaagtgggca ctaagcagac
     4741 agctccttgg catacgatta gagacacaaa cgaacaatgt acctgcctgc aatttgtacg
     4801 caaaatgtgg ctttactctc ggcggcattg acctcttcac gtataaaact agacctcaag
     4861 tctcgaacga aacagcgatg tactggtact ggttctcggg agcacaggat gacgcctaac
     4921 atatgtgaag tgtgaagggg gagattttca ctttattaga tttgtatata tgtataataa
     4981 ataaataaat aagttaaata aataattaga taagggtggt aattattact atttacaatc
     5041 aaaggtggtc ctgcacgata ccgtcgacct cgagggtatc gataccgtcg acctcgaggg
     5101 ggggcccggt accagctttt gttcccttta gtgagggtta atttcgagct tggcgtaatc
     5161 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5221 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5281 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5341 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5401 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5461 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5521 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5581 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5641 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5701 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5761 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5821 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5881 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5941 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6001 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6061 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6121 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6181 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6241 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6301 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6361 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6421 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6481 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6541 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6601 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6661 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6721 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6781 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6841 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6901 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6961 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7021 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7081 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7141 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7201 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7261 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7321 ctaagaaacc attattatca tgacattaac ctataaaaat a
//
