LOCUS       sequence-494831-        8960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of AtAFB2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8960)
  AUTHORS   Hubbe P, Sharma C, Pajonk O, Peters N, Guschtschin-Schmidt N, 
            Friemel N, Schuck S
  TITLE     An optimised system for rapid auxin-inducible protein degradation in
            budding yeast
  JOURNAL   bioRxiv 2025.09.24.678194
REFERENCE   2  (bases 1 to 8960)
  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: "bioRxiv 
            2025.09.24.678194"
COMMENT     Sequence Label: pRS405-Padh-AtAFB2
FEATURES             Location/Qualifiers
     source          1..8960
                     /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"
     CDS             complement(664..1758)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1759..2166)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2467..2922)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2604..2625)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2816..2835
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3048..3070
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3062..3079
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3063..3079
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3086..3105
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3086..3104
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3869..4573
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     primer_bind     complement(6347..6363)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      6366..6613
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6382..6400)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6655..6671)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6700..6718)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6739..6755)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6739..6755)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6752..6774)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6763..6779
                     /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(6787..6793,6794..6811,6812..6817))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6832..6853
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6970..6987)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7141..7729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7221..7240)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7900..8691,8692..8760))
                     /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     8523..8542
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8761..8865)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8933..8951
                     /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 accatatcga ctacgtcgtt aaggccgttt ctgacagagt aaaattcttg agggaacttt
      241 caccattatg ggaaatggtt caagaaggta ttgacttaaa ctccatcaaa tggtcaggtc
      301 attgagtgtt ttttatttgt tgtatttttt tttttttaga gaaaatcctc caatatataa
      361 attaggaatc atagtttcat gattttctgt tacacctaac tttttgtgtg gtgccctcct
      421 ccttgtcaat attaatgtta aagtgcaatt ctttttcctt atcacgttga gccattagta
      481 tcaatttgct tacctgtatt cctttacatc ctcctttttc tccttcttga taaatgtatg
      541 tagattgcgt atatagtttc gtctacccta tgaacatatt ccattttgta atttcgtgtc
      601 gtttctatta tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aaagagaatc
      661 tttttaagca aggattttct taacttcttc ggcgacagca tcaccgactt cggtggtact
      721 gttggaacca cctaaatcac cagttctgat acctgcatcc aaaacctttt taactgcatc
      781 ttcaatggcc ttaccttctt caggcaagtt caatgacaat ttcaacatca ttgcagcaga
      841 caagatagtg gcgatagggt tgaccttatt ctttggcaaa tctggagcag aaccgtggca
      901 tggttcgtac aaaccaaatg cggtgttctt gtctggcaaa gaggccaagg acgcagatgg
      961 caacaaaccc aaggaacctg ggataacgga ggcttcatcg gagatgatat caccaaacat
     1021 gttgctggtg attataatac catttaggtg ggttgggttc ttaactagga tcatggcggc
     1081 agaatcaatc aattgatgtt gaaccttcaa tgtagggaat tcgttcttga tggtttcctc
     1141 cacagttttt ctccataatc ttgaagaggc caaaacatta gctttatcca aggaccaaat
     1201 aggcaatggt ggctcatgtt gtagggccat gaaagcggcc attcttgtga ttctttgcac
     1261 ttctggaacg gtgtattgtt cactatccca agcgacacca tcaccatcgt cttcctttct
     1321 cttaccaaag taaatacctc ccactaattc tctgacaaca acgaagtcag tacctttagc
     1381 aaattgtggc ttgattggag ataagtctaa aagagagtcg gatgcaaagt tacatggtct
     1441 taagttggcg tacaattgaa gttctttacg gatttttagt aaaccttgtt caggtctaac
     1501 actaccggta ccccatttag gaccacccac agcacctaac aaaacggcat cagccttctt
     1561 ggaggcttcc agcgcctcat ctggaagtgg aacacctgta gcatcgatag cagcaccacc
     1621 aattaaatga ttttcgaaat cgaacttgac attggaacga acatcagaaa tagctttaag
     1681 aaccttaatg gcttcggctg tgatttcttg accaacgtgg tcacctggca aaacgacgat
     1741 cttcttaggg gcagacatta gaatggtata tccttgaaat atatatatat atattgctga
     1801 aatgtaaaag gtaagaaaag ttagaaagta agacgattgc taaccaccta ttggaaaaaa
     1861 caataggtcc ttaaataata ttgtcaactt caagtattgt gatgcaagca tttagtcatg
     1921 aacgcttctc tattctatat gaaaagccgg ttccggcgct ctcacctttc ctttttctcc
     1981 caatttttca gttgaaaaag gtatatgcgt caggcgacct ctgaaattaa caaaaaattt
     2041 ccagtcatcg aatttgattc tgtgcgatag cgcccctgtg tgttctcgtt atgttgagga
     2101 aaaaaataat ggttgctaag agattcgaac tcttgcatct tacgatacct gagtattccc
     2161 acagttaact gcggtcaaga tatttcttga atcaggcgcc ttagaccgct cggccaaaca
     2221 accaattact tgttgagaaa tagagtataa ttatcctata aatataacgt ttttgaacac
     2281 acatgaacaa ggaagtacag gacaattgat tttgaagaga atgtggattt tgatgtaatt
     2341 gttgggattc catttttaat aaggcaataa tattaggtat gtagatatac tagaagttct
     2401 cctcgaccgg tcgatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     2461 atcaggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2521 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2581 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2641 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2701 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2761 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2821 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2881 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct
     2941 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     3001 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     3061 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcgc
     3121 cgggatcgaa gaaatgatgg taaatgaaat aggaaatcaa ggagcatgaa ggcaaaagac
     3181 aaatataagg gtcgaacgaa aaataaagtg aaaagtgttg atatgatgta tttggctttg
     3241 cggcgccgaa aaaacgagtt tacgcaattg cacaatcatg ctgactctgt ggcggacccg
     3301 cgctcttgcc ggcccggcga taacgctggg cgtgaggctg tgcccggcgg agttttttgc
     3361 gcctgcattt tccaaggttt accctgcgct aaggggcgag attggagaag caataagaat
     3421 gccggttggg gttgcgatga tgacgaccac gacaactggt gtcattattt aagttgccga
     3481 aagaacctga gtgcatttgc aacatgagta tactagaaga atgagccaag acttgcgaga
     3541 cgcgagtttg ccggtggtgc gaacaataga gcgaccatga ccttgaaggt gagacgcgca
     3601 taaccgctag agtactttga agaggaaaca gcaatagggt tgctaccagt ataaatagac
     3661 aggtacatac aacactggaa atggttgtct gtttgagtac gctttcaatt catttgggtg
     3721 tgcactttat tatgttacaa tatggaaggg aactttacac ttctcctatg cacatatatt
     3781 aattaaagtc caatgctagt agagaagggg ggtaacaccc ctccgcgctc ttttccgatt
     3841 tttttctaaa ccgtggaata tttcggatat ccttttgttg tttccgggtg tacaatatgg
     3901 acttcctctt ttctggcaac caaacccata catcgggatt cctataatac cttcgttggt
     3961 ctccctaaca tgtaggtggc ggaggggaga tatacaatag aacagatacc agacaagaca
     4021 taatgggcta aacaagacta caccaattac actgcctcat tgatggtggt acataacgaa
     4081 ctaatactgt agccctagac ttgatagcca tcatcatatc gaagtttcac tacccttttt
     4141 ccatttgcca tctattgaag taataatagg cgcatgcaac ttcttttctt tttttttctt
     4201 ttctctctcc cccgttgttg tctcaccata tccgcaatga caaaaaaatg atggaagaca
     4261 ctaaaggaaa aaattaacga caaagacagc accaacagat gtcgttgttc cagagctgat
     4321 gaggggtatc tcgaagcaca cgaaactttt tccttccttc attcacgcac actactctct
     4381 aatgagcaac ggtatacggc cttccttcca gttacttgaa tttgaaataa aaaaaagttt
     4441 gctgtcttgc tatcaagtat aaatagacct gcaattatta atcttttgtt tcctcgtcat
     4501 tgttctcgtt ccctttcttc cttgtttctt tttctgcaca atatttcaag ctataccaag
     4561 catacaatca actccaagct ggccgctcta gaactagtat gaactacttt cccgatgagg
     4621 tcattgagca cgtctttgat ttcgtcacaa gccacaagga taggaacgcc attagcctgg
     4681 tctgtaagag ctggtacaag atcgagcggt attccagaca gaaggtgttc atcggcaact
     4741 gctacgccat caatccagag aggctgctgc ggagatttcc ctgtctgaag agcctgaccc
     4801 tgaagggcaa gccccacttc gccgacttta acctggtgcc tcacgagtgg ggaggattcg
     4861 tgctgccatg gatcgaggcc ctggcacggt ccagagtggg cctggaggag ctgaggctga
     4921 agcgcatggt ggtgacagac gagtctctgg agctgctgtc tcggagcttc gtgaacttca
     4981 agagcctggt gctggtgtcc tgcgagggct ttaccacaga tggcctggca agcatcgcag
     5041 caaactgtag gcacctgcgg gacctggacc tgcaggagaa tgagatcgac gatcacagag
     5101 gccagtggct gtcctgcttc cccgatacct gtaccacact ggtgacactg aactttgcct
     5161 gcctggaggg cgagacaaat ctggtggccc tggagaggct ggtggcacgc tctcccaacc
     5221 tgaagagcct gaagctgaat agggcagtgc ctctggacgc actggcaaga ctgatggcat
     5281 gcgcaccaca gatcgtggat ctgggcgtgg gctcctacga gaacgaccct gattccgagt
     5341 cttatctgaa gctgatggcc gtgatcaaga agtgtaccag cctgcgcagc ctgtccggct
     5401 tcctggaggc agcacctcac tgcctgtccg cctttcaccc aatctgtcac aacctgacat
     5461 ccctgaatct gtcttacgcc gccgagatcc acggctccca cctgatcaag ctgatccagc
     5521 actgcaagaa gctgcagagg ctgtggatcc tggactccat cggcgataag ggcctggagg
     5581 tggtggcctc tacctgtaag gagctgcagg agctgcgcgt gttcccatct gacctgctgg
     5641 gaggaggaaa caccgcagtg acagaggagg gcctggtggc aatctctgcc ggatgcccca
     5701 agctgcacag catcctgtat ttttgtcagc agatgaccaa cgccgccctg gtgacagtgg
     5761 ccaagaactg ccccaatttc atcaggtttc gcctgtgcat cctggagccc aataagcctg
     5821 accacgtgac atctcagcct ctggatgagg gcttcggcgc catcgtgaag gcctgcaaga
     5881 gcctgaggcg cctgtctctg agcggcctgc tgaccgacca ggtgttcctg tacatcggca
     5941 tgtatgccaa ccagctggag atgctgtcta tcgcctttgc cggcgacaca gataagggca
     6001 tgctgtacgt gctgaatggc tgtaagaaga tgaagaagct ggagatcaga gacagccctt
     6061 ttggcgatac cgccctgctg gcagacgtga gcaagtatga gacaatgcgg agcctgtgga
     6121 tgagctcctg cgaggtgacc ctgagcggct gtaagagact ggccgagaag gccccatggc
     6181 tgaacgtgga gatcatcaac gagaatgaca acaataggat ggaggagaat ggccacgagg
     6241 gccgccagaa ggtggataag ctgtatctgt ataggaccgt cgtcgggact cggatggatg
     6301 cccccccctt tgtctggatt ctgtaagaat tcgatatcaa gcttatcgat accgtcgacc
     6361 tcgagtcatg taattagtta tgtcacgctt acattcacgc cctcccccca catccgctct
     6421 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt tttttatagt
     6481 tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacagacg
     6541 cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg gacgctcgaa
     6601 ggctttaatt tgcggccggt accccgggct gcaggaattc gatatcaagc ttatcgatac
     6661 cgtcgacctc gagggggggc ccggtaccag cttttgttcc ctttagtgag ggttaatttc
     6721 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6781 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6841 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6901 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6961 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     7021 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7081 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7141 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7201 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7261 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7321 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7381 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7441 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7501 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7561 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     7621 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7681 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7741 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7801 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7861 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7921 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7981 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     8041 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     8101 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8161 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     8221 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8281 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8341 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8401 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8461 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8521 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8581 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8641 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8701 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8761 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8821 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8881 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     8941 tatcacgagg ccctttcgtc
//
