LOCUS       sequence-402962-        9864 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of OsTIR1(F74A)-3xMyc at NEUT5L locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9864)
  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 9864)
  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: pHLP712
FEATURES             Location/Qualifiers
     source          1..9864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    69..572
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        609..713
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(714..782,783..1574)
                     /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     complement(932..951)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1745..2333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2234..2253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2487..2504
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2621..2642
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2657..2662,2663..2680,2681..2687)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     CDS             7267..7296
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             7303..7332
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             7339..7368
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      7394..7591
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(7663..7682)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7664..7682)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7689..7706)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7689..7705)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7698..7720)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7933..7952)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8458..8479
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(8719..9393)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(9394..9674)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(9695..9714)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9814..9836
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgga tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa
      121 tttgggaatt tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta
      181 aataaagaag gtagaagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa
      241 tttcaacaaa aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag
      301 atatacattc gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt
      361 ctacacagac aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa
      421 aaggtagtat ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta
      481 ttttttcttt aatttctttt tttactttct atttttaatt tatatattta tattaaaaaa
      541 tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc acttttcggg
      601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
      961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1861 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2401 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2461 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2521 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     2581 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     2641 attaggcacc ccaggcttta cactttatgc ttccggctcc tatgttgggc cggctaggcc
     2701 tgtaattgta gtaagaatga caagtatcag ttgagaaaaa aagtaacaaa gagaaagctc
     2761 ggaggaggct ccccaaaggt tttatcacca atggtggtac cactaaccca agaacagaaa
     2821 aagcattgat attttgaaaa aatacaatac tcatctttta acccgaacct tttcttgtcc
     2881 tttaaaactt ttatctcatg tctgtccaaa gtcatattgt cttactactt tctctccttc
     2941 atcgtccttc cggccccccc ggatccggat ggtataaacg gaaacaaaaa aaagagctgg
     3001 tactactttc tttaaaatta ttttattatt tgattttatt taatagtata tattatattt
     3061 tgaacgtaga ttattttgtt gaaagttgct gtagtgccat tgattcgtaa cactaattct
     3121 gtattagtca ttcctcttgt ttgatagtat ccaaaaaaac ggctattttt ttgcaatctt
     3181 atttcctgca tattatacag ataacataat gaaagaaaaa atcttttttt ttgttcttca
     3241 atgatgattt caaccattct tttaaacatt gatcaattcc tgagcaacaa ccccatacac
     3301 actggtttat atatcgcccc ttttacagtt gaagaaagaa atagaaatag aaatagcaaa
     3361 caaaagatat gacagtcaac actaagacct atagtgagag agcagaaact catgcctcac
     3421 cagtagcaca acgattattt cgattaatgg aactgaagaa aaccaattta tgtgcatcaa
     3481 ttgatgttga taccactaag gaattccttg aattaattga taaattgggt ccttatgtat
     3541 gcttaatcaa gactcatatt gatataatca atgatttttc ctatgaatcc actattgaac
     3601 cattattaga actttcacgt aaacatcaat ttatgatttt tgaagataga aaatttgctg
     3661 atattggtaa taccgtgaag aaacaatata ttggtggagt ttataaaatt agtagttggg
     3721 cagatattac taatgctcat ggtgtcactg ggaatggagt agttgaagga ttaaaacagg
     3781 gagctaaaga aaccaccacc aaccaagagc caagagggtt attgatgtta gctgaattat
     3841 catcagtggg atcattagca tatggagaat attctcaaaa aactgttgaa attgctaaat
     3901 ccgataagga atttgttgtt ggatttattg cccaacgtga tatgggtgga caagaagaag
     3961 gatttgattg gcttattatg acacctggag ttggattaga tgataaaggt gatggattag
     4021 gacaacaata tagaactgtt gatgaagttg ttagcactgg aactgatatt atcattgttg
     4081 gtagaggatt gtttggtaaa ggaagagatc cagatattga aggtaaaagg tatagagatg
     4141 ctggttggaa tgcttatttg aaaaaaactg gccaattata aatgtgaagg gggagatttt
     4201 cactttatta gatttgtata tatgtataat aaataaataa ataagttaaa taaataatta
     4261 aataagggtg gtaattatta ctatttacaa tcaaaggtgg tccttctagt cgagcgtcaa
     4321 aactagagaa taataaagaa aacgatcttt tcaaaaagaa aaaacctttt agttttcctt
     4381 tgttgttgtt gtgggtgtgt gctatttata ttatatagtt tactcataat accataaaat
     4441 attcggtttg attaggttat tttaataagc taatttgttt ctaatcgtgt aatttatgct
     4501 gtgtatatta agtagtgtgt gcactgccca aaaatgtttg ttgtttatag tcggttaaag
     4561 agaaaaaaga aaaaaagatc catacacaca cgttaattag ttgttcaacg taatacactc
     4621 atattttgtt cttatttgct ttcggtcgct gttctcacca agatttattg ccaacgaaac
     4681 aatttttttt tatatatttt cagatttttc tttttttcct ttcctttcct tttctaattt
     4741 tcactcctgg ttttctttct ttcttagaaa cattatctcg atattaatat taaaaaaata
     4801 taatcattca aaatggacgg tggtatgttt tagtttagct tcaattctaa ttgattgatt
     4861 aatcagttga ttggtttcaa tatgacaaat gggtagggtg ggaaaacttc attttcaatt
     4921 cagatcaaac ttttttgttg tcgacataat atttctcgtt tgggatgtta ctgtcacatt
     4981 aataatacac acacatcagc ttataatttt gaaagtaatt tatcagatat gttgtgacga
     5041 tcagtggaaa tggctaactt caatgtatct gttcttcccc tttttcaaag ttcacgtttt
     5101 ttgattgatt gattgatctg tcggcagtgg tttcaaaacc attcggtgag taatcctatc
     5161 aatcaatgtt acgacaaaag gctcaatatt caaaattgca atgttttatg ttttcctacg
     5221 tgtacttgtg caaggcaatt gattcaacat tgcttttggt gtttgacgag tttctagttt
     5281 ggacttgtgt tgttatctgg gctatacaga tttcccggct cactatgaat tttttttttc
     5341 gacgctcagt gcacacaact ataaacaaca caaacacaaa cacagcaaga aaaaaaaaaa
     5401 acgaacattg aattgaaacc aagccaactg aaaaattcct tatttaaatg actgtcatac
     5461 taacccattt ttatagaaga agttgctgct ttagttatcg ataacggttc tcatatgaca
     5521 tacttccctg aagaagtggt agagcatatt ttctcctttt tacctgccca aagagatcga
     5581 aataccgtct cacttgtatg caaagtttgg tatgagattg aacgattatc aagacgaggg
     5641 gtctttgtag gtaattgtta tgcagtgcgt gccggtcgag tcgctgctag attccctaac
     5701 gtgcgtgccc ttaccgtcaa gggtaagcct cacgctgcag attttaattt agtgccacct
     5761 gattggggtg gatatgccgg tccatggatc gaagccgccg ctcgtggttg tcacggattg
     5821 gaagaattac gaatgaagcg aatggtcgtt tccgacgaat ccttagaatt gttagcaaga
     5881 agttttccac gattcagagc attagtatta atatcatgcg agggtttttc aactgacgga
     5941 ttagccgccg tagcctcaca ttgcaagtta ttgagagaat tggacttaca agaaaatgag
     6001 gtcgaggacc gtggacctcg atggttgtca tgctttcctg attcctgcac atcacttgtg
     6061 tccttgaatt tcgcatgtat caagggggaa gttaacgctg ggtccttgga aagattggtg
     6121 tcacgttcac ctaatttacg ttctttgaga ttgaaccgtt ccgtatccgt tgatacttta
     6181 gccaaaattt tgcttagaac tcctaatttg gaggatcttg gaactggaaa tttgactgat
     6241 gacttccaaa cagagtctta ttttaagtta acatccgctt tggaaaagtg taagatgtta
     6301 cgaagtttat ccggtttttg ggacgcatct ccagtgtgct tgtcatttat ttatcctttg
     6361 tgcgcccagt taactgggtt aaatttatct tatgccccta ccttggatgc ttcagatctt
     6421 actaaaatga taagtcgatg tgtcaaatta cagcgattat gggtcttaga ctgtatctct
     6481 gacaaggggt tacaagtagt agcctctagt tgcaaagatc ttcaagaatt aagagtgttt
     6541 cctagtgact tctacgtggc aggttattca gcagtaacag aggagggatt ggtcgctgta
     6601 tcattaggtt gcccaaagtt gaattcattg ttatacttct gccaccaaat gactaatgcc
     6661 gcattggtca ctgttgccaa gaattgccct aactttactc gttttcgttt gtgtatattg
     6721 gagccaggta agccagacgt tgtcactagt caaccattgg atgagggatt cggtgcaatt
     6781 gtaagagaat gtaaggggtt gcaacgtttg agtatcagtg ggttattaac agacaaggtt
     6841 ttcatgtata ttggtaaata cgccaagcag cttgaaatgt taagtatagc attcgctgga
     6901 gatagtgata agggtatgat gcacgtaatg aatgggtgta aaaatcttcg aaaattagaa
     6961 atccgagatt caccttttgg agacgctgcc cttcttggga atttcgctcg atacgaaacc
     7021 atgagatccc tttggatgtc ctcatgcaac gtgacattga agggatgcca agttcttgcc
     7081 tccaagatgc ctatgttaaa cgtagaggtt attaatgaga gagatgggtc caacgagatg
     7141 gaagaaaacc acggagattt gccaaaagtt gagaagcttt atgtataccg taccactgca
     7201 ggggctcgtg atgatgctcc aaattttgtc aagatattgg ttgatagtgg tagtgcagct
     7261 agtggggagc aaaaattgat ttctgaagaa gatttaaatg gtgagcaaaa gttgatatca
     7321 gaggaagact tgaatggtga acagaaatta atctccgaag aagacttaaa tggttcatcc
     7381 taatcgagtc agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata
     7441 gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt
     7501 tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt
     7561 caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc
     7621 agtgtcgacg tttaaacgag ctcgaattcg gtacccaatt cgccctatag tgagtcgtat
     7681 tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     7741 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     7801 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg
     7861 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     7921 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggtaaac
     7981 aagtggtatt caagcacaat tcagttttct ggatctctaa taattgcaat tgcaattgct
     8041 tcacatactc tgtttataat tcagtttctt tctcttgatg ttactgatat attaagttaa
     8101 taaataccaa taactttttt tcaaaaatct tctattttcc tttcaattga ttatttaaac
     8161 acgaatcgtt aataagctgt gattgctttt tctttttctt tttcttttct ttgttcaagt
     8221 ttgaaaaaag taaaaaaaaa ggtttctctt gttcgatgta gaattcagtg ggttaaggcc
     8281 tagccggccg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     8341 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     8401 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     8461 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     8521 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     8581 acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
     8641 gcatctgtgc ggtatttcac accgcataga tcggcaagtg cacaaacaat acttaaataa
     8701 atactactca gtaataacct atttcttagc atttttgacg aaatttgcta ttttgttaga
     8761 gtcttttaca ccatttgtct ccacacctcc gcttacatca acaccaataa cgccatttaa
     8821 tctaagcgca tcaccaacat tttctggcgt cagtccacca gctaacataa aatgtaagct
     8881 ttcggggctc tcttgccttc caacccagtc agaaatcgag ttccaatcca aaagttcacc
     8941 tgtcccacct gcttctgaat caaacaaggg aataaacgaa tgaggtttct gtgaagctgc
     9001 actgagtagt atgttgcagt cttttggaaa tacgagtctt ttaataactg gcaaaccgag
     9061 gaactcttgg tattcttgcc acgactcatc tccatgcagt tggacgatat caatgccgta
     9121 atcattgacc agagccaaaa catcctcctt aggttgatta cgaaacacgc caaccaagta
     9181 tttcggagtg cctgaactat ttttatatgc ttttacaaga cttgaaattt tccttgcaat
     9241 aaccgggtca attgttctct ttctattggg cacacatata atacccagca agtcagcatc
     9301 ggaatctaga gcacattctg cggcctctgt gctctgcaag ccgcaaactt tcaccaatgg
     9361 accagaacta cctgtgaaat taataacaga catactccaa gctgcctttg tgtgcttaat
     9421 cacgtatact cacgtgctca atagtcacca atgccctccc tcttggccct ctccttttct
     9481 tttttcgacc gaattaattc ttaatcggca aaaaaagaaa agctccggat caagattgta
     9541 cgtaaggtga caagctattt ttcaataaag aatatcttcc actactgcca tctggcgtca
     9601 taactgcaaa gtacacatat attacgatgc tgttctatta aatgcttcct atattatata
     9661 tatagtaatg tcgtgatcta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     9721 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     9781 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     9841 caccgtcatc accgaaacgc gcga
//
