LOCUS       sequence-190867-        6018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  S. cerevisiae expression of F-box protein AFB2 under control of the 
            ADH1 promoter, TRP1 marker (for use with the AID degron system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6018)
  TITLE     Unpublished Ulrich lab AID plasmids
REFERENCE   2  (bases 1 to 6018)
  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"
COMMENT     Sequence Label: YIp204-PADH1-AFB2
FEATURES             Location/Qualifiers
     source          1..6018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..704
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     protein_bind    719..743
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2478..2502)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2539..2555)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2539..2555)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2552..2574)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2563..2579
                     /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(2587..2593,2594..2611,2612..2617))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2632..2653
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2770..2787)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2941..3529)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3021..3040)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3700..4491,4492..4560))
                     /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     4323..4342
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4561..4665)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4733..4751
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4753..4854
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             4855..5529
                     /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"
     primer_bind     complement(5648..5670)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5770..5789
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5983..6005
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5997..6014
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5998..6014
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tccttttgtt gtttccgggt gtacaatatg gacttcctct tttctggcaa ccaaacccat
       61 acatcgggat tcctataata ccttcgttgg tctccctaac atgtaggtgg cggaggggag
      121 atatacaata gaacagatac cagacaagac ataatgggct aaacaagact acaccaatta
      181 cactgcctca ttgatggtgg tacataacga actaatactg tagccctaga cttgatagcc
      241 atcatcatat cgaagtttca ctaccctttt tccatttgcc atctattgaa gtaataatag
      301 gcgcatgcaa cttcttttct ttttttttct tttctctctc ccccgttgtt gtctcaccat
      361 atccgcaatg acaaaaaaat gatggaagac actaaaggaa aaaattaacg acaaagacag
      421 caccaacaga tgtcgttgtt ccagagctga tgaggggtat ctcgaagcac acgaaacttt
      481 ttccttcctt cattcacgca cactactctc taatgagcaa cggtatacgg ccttccttcc
      541 agttacttga atttgaaata aaaaaaagtt tgctgtcttg ctatcaagta taaatagacc
      601 tgcaattatt aatcttttgt ttcctcgtca ttgttctcgt tccctttctt ccttgtttct
      661 ttttctgcac aatatttcaa gctataccaa gcatacaatc aactccggat cccccatcac
      721 aagtttgtac aaaaaagcag gcttcaaaat gaattatttc ccagatgaag taatagagca
      781 tgtattcgac tttgtaacat ctcacaaaga caggaatgct atatctcttg tatgcaaatc
      841 atggtacaag attgaaagat acagtaggca aaaggttttc attggaaact gttatgccat
      901 taatccagag aggttgcttc ggagattccc atgtctaaag tctttgactt tgaaaggaaa
      961 acctcatttt gcggatttca atttggttcc tcatgaatgg ggaggttttg tgctaccttg
     1021 gattgaggct ttggctagaa gccgtgtagg acttgaagag cttaggttga agaggatggt
     1081 tgttactgat gagagtcttg agctgctttc tcgttctttt gtcaatttta agtctttggt
     1141 ccttgttagc tgtgaaggtt ttaccactga tggtcttgcc tctattgccg ctaattgcag
     1201 gcatcttcgg gatcttgatt tgcaagagaa tgaaatcgat gatcatagag gtcaatggtt
     1261 aagttgtttc ccagacactt gcacgactct tgtcacgcta aactttgcgt gcctcgaagg
     1321 agaaactaat ctggttgctc tagagaggct tgttgctagg tctccaaacc taaagagtct
     1381 gaagctaaat cgtgcagtac cgttagatgc actcgcaagg ttaatggcgt gtgcgccgca
     1441 gatagttgac ttaggagtag ggtcttatga gaatgaccca gattccgagt cttacttgaa
     1501 actcatggct gtcataaaga aatgcacctc gttgaggagt ttgtcgggtt ttctagaggc
     1561 tgctcctcac tgtctctcag ctttccaccc aatatgtcat aacctcacct ccttgaatct
     1621 tagttacgca gctgagattc atggtagcca ccttattaag cttattcagc attgcaagaa
     1681 acttcagcgg ttatggattt tggatagtat aggtgacaaa gggcttgaag ttgtagcttc
     1741 tacatgtaaa gagttacaag agcttagggt ttttccatct gatttactcg gtggaggcaa
     1801 cacagctgtg accgaagaag gtctagttgc catctcggca ggctgcccta agctccactc
     1861 tatactctac ttctgccaac aaatgacaaa cgcagctctc gtaaccgttg ccaagaactg
     1921 tccaaatttc atccgtttcc gactctgcat cctcgagcca aacaaacccg atcacgtcac
     1981 atctcaacct ctagacgaag gctttggagc aatcgtcaaa gcctgcaaga gcctgagaag
     2041 gctttctctc tcaggtctcc ttacagacca agtcttcctc tacatcggaa tgtacgcgaa
     2101 tcagctcgag atgctctcca tagcctttgc aggagataca gacaaaggca tgctatatgt
     2161 gttgaatggt tgcaaaaaga tgaagaaact agagataagg gatagtccgt ttggggacac
     2221 ggcgcttctt gctgatgtga gcaagtatga aacaatgcga tccctttgga tgtcttcatg
     2281 tgaagtcaca ctcagtggat gcaaaaggct cgcagagaaa gcgccatggc tcaatgtaga
     2341 gatcataaac gagaatgata ataaccggat ggaagaaaac ggacacgagg ggaggcagaa
     2401 agtggataag ttgtatctgt accggactgt ggttgggaca agaatggatg cgccgccatt
     2461 tgtgtggatt ctctaacacc cagctttctt gtacaaagtg gtgatgggct gcaggcatgc
     2521 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2581 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2641 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2701 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2761 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2821 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2881 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2941 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3001 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3061 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3121 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3181 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3241 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3301 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3361 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3421 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3481 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3541 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3601 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3661 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3721 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3781 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3841 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3901 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3961 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4021 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4081 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4141 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4201 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4261 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4321 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4381 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4441 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4501 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4561 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4621 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4681 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     4741 tatcacgagg ccaattcggt cgaaaaaaga aaaggagagg gccaagaggg agggcattgg
     4801 tgactattga gcacgtgagt atacgtgatt aagcacacaa aggcagcttg gagtatgtct
     4861 gttattaatt tcacaggtag ttctggtcca ttggtgaaag tttgcggctt gcagagcaca
     4921 gaggccgcag aatgtgcact agattccgat gctgacttgc tgggtattat atgtgtgccc
     4981 aatagaaaga gaacaattga cccggttatt gcaaggaaaa tttcaagtct tgtaaaagca
     5041 tataaaaata gttcaggcac tccgaaatac ttggttggcg tgtttcgtaa tcaacctaag
     5101 gaggatgttt tggctctggt caatgattac ggcattgata tcgtccaact gcatggagat
     5161 gagtcgtggc aagaatacca agagttcctc ggtttgccag ttattaaaag actcgtattt
     5221 ccaaaagact gcaacatact actcagtgca gcttcacaga aacctcattc gtttattccc
     5281 ttgtttgatt cagaagcagg tgggacaggt gaacttttgg attggaactc gatttctgac
     5341 tgggttggaa ggcaagagag ccccgagagc ttacatttta tgttagctgg tggactgacg
     5401 ccagaaaatg ttggtgatgc gcttagatta aatggcgtta ttggtgttga tgtaagcgga
     5461 ggtgtggaga caaatggtgt aaaagactct aacaaaatag caaatttcgt caaaaatgct
     5521 aagaaatagg ttattactga gtagtattta tttaagtatt gtttgtgcac ttgcctgcaa
     5581 gccttttgaa aagcaagcat aaaagatcgc cctttcgtct cgcgcgtttc ggtgatgacg
     5641 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     5701 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     5761 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     5821 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     5881 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     5941 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     6001 aaacgacggc cagtgaat
//
