LOCUS       sequence-494868-        9265 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 9265)
  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 9265)
  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: pNH605-Padh-AtAFB2(F74A)
FEATURES             Location/Qualifiers
     source          1..9265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        276..683
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     promoter        2509..3213
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     primer_bind     complement(4980..4996)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4981..4997)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4999..5246
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5015..5033)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5812..5831)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5813..5831)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5838..5855)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5838..5854)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5847..5869)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5995..6450
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6082..6101)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6292..6313
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6806..6825)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6925..6947
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6985..7003)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7088..7192
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7193..7261,7262..8053)
                     /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(7411..7430)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8224..8812
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8713..8732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8966..8983
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9100..9121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9136..9141,9142..9159,9160..9166)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9174..9190
                     /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)."
     primer_bind     9179..9201
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9198..9214
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9198..9214
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        9235..9253
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggccggccat ttaaatgttt aaacggccac gctggccgag gagaacttct agtatatcta
       61 catacctaat attattgcct tattaaaaat ggaatcccaa cgattacatc aaaatccaca
      121 ttctcttcaa aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa aacgttatat
      181 ttataggata attatactct atttctcaac aagtaattgg ttgtttggcc gagcggtcta
      241 aggcgcctga ttcaagaaat atcttgaccg cagttaactg tgggaatact caggtatcgt
      301 aagatgcaag agttcgaatc tcttagcaac cattattttt ttcctcaaca taacgagaac
      361 acacaggggc gctatcgcac agaatcaaat tcgatgactg gaaatttttt gttaatttca
      421 gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga
      481 gagcgccgga accggctttt catatagaat agagaagcgt tcatgactaa atgcttgcat
      541 cacaatactt gaagttgaca atattattta aggacctatt gttttttcca ataggtggtt
      601 agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata tatatatata
      661 tatttcaagg atataccatt ctaatgtctg cccctaagtt accaatagat agcggtttaa
      721 tataaggttt caacaagaaa tattttgtga aaatcaaaag ttaatagttt cactgtcgaa
      781 aatatgaaag tttaagtttt cagttcatcc cattgtagtt atcaaatttc tgtgtttccc
      841 gtatatgtgt gctctgtatt catcgctatc ttgaaactgg ttatgatcgt tgcgcatgtg
      901 acctgtatcg ctcaaaagag cccggcaccg gtaccgggca aaagccccta gcaagctaaa
      961 atgattcaac tttcgtagta atttgttcaa ccggtcacag taaagtcaat gactaacttt
     1021 tgaatcgagg atgtatttcc aattgaaagc aatgtatata taaaccattg tgttgaagct
     1081 tgtcttacag ttttcttgta tcctcccatt gttcatatag tacttaaagt accataatcc
     1141 ttaaagaaca aacttgaaag agaatacact atacacaact aataaataaa gaataatcat
     1201 ggctgtgacc aagacaattg tagttctacc aggtgaccat gttggtcaag aaatcactga
     1261 agaggccatt aaagttttaa atgctattca ggaatgtcgt ccagacaagg tcaatttcaa
     1321 gtttgatcat catttgatcg gtggtgctgc aattgatgcc actggtgttc cattaccaga
     1381 cgaagctttg gaggcctcta agaaagctga tgccgtgctt ctgggtgctg ttggtggtcc
     1441 aaaatggggt actggcgctg tcagaccaga acaaggtctt ttgaaaatcc gtaaggagtt
     1501 gcaattgtat gctaatctaa gaccatgtaa ttttgcatct gattccttac tagatctatc
     1561 gccattgaag cctgaaattg caagaggtac agatttcgtc gttgttagag aactagtggg
     1621 tggtatttat ttcggggaga gaaaagaaga tgaaggtgat ggtgtcgcct gggatagcga
     1681 aaagtattct gtgcccgaag ttcaaagaat cactagaatg gctgctttca tggctctgca
     1741 acacaaccca cctttaccta tttggtctct agataaagcc aacgttctag catcatcaag
     1801 attatggaga aaaacagtgg aagagacgat taagaatgaa ttcccacaat tgactgttca
     1861 acatcaattg attgattccg ctgctatgat cttggttaag aacccaactc atctaaatgg
     1921 tattattatc acaaacaata tgtttggtga tattatctcc gatgaagcct ccgtgatccc
     1981 aggttcgtta ggattacttc catctgcctc cttggcatct cttccagata agaatacggc
     2041 atttggttta tatgagccat gtcatggctc tgctccagac ttaccaaagg gaaaggttaa
     2101 ccccgtagca acaatcctat ctgctgctat gatgttgaag ttgtctctgg acctgttcga
     2161 agaaggtgaa attatagaac aagcagtcaa gaaagtgttg gattctggta tcagaacagc
     2221 ggatctaaag ggtaccaact ctactactga ggtcggtgat gcagtagcga aagctgtcag
     2281 ggaactatta gcttagactt agtagatgta cagttgcata cataagtctg aacagaaaga
     2341 agtaaatatc atatctagtt cggtttagaa atcagtgact atattggttt tgaccagatc
     2401 gcaatccttg caagcattta gtaatgacta tcaaaccggt gaatcatctg ataaggctat
     2461 attagcttgt gcattcgcat gtatcgggaa acgaacggta ccgggcccat ccttttgttg
     2521 tttccgggtg tacaatatgg acttcctctt ttctggcaac caaacccata catcgggatt
     2581 cctataatac cttcgttggt ctccctaaca tgtaggtggc ggaggggaga tatacaatag
     2641 aacagatacc agacaagaca taatgggcta aacaagacta caccaattac actgcctcat
     2701 tgatggtggt acataacgaa ctaatactgt agccctagac ttgatagcca tcatcatatc
     2761 gaagtttcac tacccttttt ccatttgcca tctattgaag taataatagg cgcatgcaac
     2821 ttcttttctt tttttttctt ttctctctcc cccgttgttg tctcaccata tccgcaatga
     2881 caaaaaaatg atggaagaca ctaaaggaaa aaattaacga caaagacagc accaacagat
     2941 gtcgttgttc cagagctgat gaggggtatc tcgaagcaca cgaaactttt tccttccttc
     3001 attcacgcac actactctct aatgagcaac ggtatacggc cttccttcca gttacttgaa
     3061 tttgaaataa aaaaaagttt gctgtcttgc tatcaagtat aaatagacct gcaattatta
     3121 atcttttgtt tcctcgtcat tgttctcgtt ccctttcttc cttgtttctt tttctgcaca
     3181 atatttcaag ctataccaag catacaatca actccaagct ggccgctcta gaactagtat
     3241 gaactacttt cccgatgagg tcattgagca cgtctttgat ttcgtcacaa gccacaagga
     3301 taggaacgcc attagcctgg tctgtaagag ctggtacaag atcgagcggt attccagaca
     3361 gaaggtgttc atcggcaact gctacgccat caatccagag aggctgctgc ggagatttcc
     3421 ctgtctgaag agcctgaccc tgaagggcaa gccccacgcc gccgacttta acctggtgcc
     3481 tcacgagtgg ggaggattcg tgctgccatg gatcgaggcc ctggcacggt ccagagtggg
     3541 cctggaggag ctgaggctga agcgcatggt ggtgacagac gagtctctgg agctgctgtc
     3601 tcggagcttc gtgaacttca agagcctggt gctggtgtcc tgcgagggct ttaccacaga
     3661 tggcctggca agcatcgcag caaactgtag gcacctgcgg gacctggacc tgcaggagaa
     3721 tgagatcgac gatcacagag gccagtggct gtcctgcttc cccgatacct gtaccacact
     3781 ggtgacactg aactttgcct gcctggaggg cgagacaaat ctggtggccc tggagaggct
     3841 ggtggcacgc tctcccaacc tgaagagcct gaagctgaat agggcagtgc ctctggacgc
     3901 actggcaaga ctgatggcat gcgcaccaca gatcgtggat ctgggcgtgg gctcctacga
     3961 gaacgaccct gattccgagt cttatctgaa gctgatggcc gtgatcaaga agtgtaccag
     4021 cctgcgcagc ctgtccggct tcctggaggc agcacctcac tgcctgtccg cctttcaccc
     4081 aatctgtcac aacctgacat ccctgaatct gtcttacgcc gccgagatcc acggctccca
     4141 cctgatcaag ctgatccagc actgcaagaa gctgcagagg ctgtggatcc tggactccat
     4201 cggcgataag ggcctggagg tggtggcctc tacctgtaag gagctgcagg agctgcgcgt
     4261 gttcccatct gacctgctgg gaggaggaaa caccgcagtg acagaggagg gcctggtggc
     4321 aatctctgcc ggatgcccca agctgcacag catcctgtat ttttgtcagc agatgaccaa
     4381 cgccgccctg gtgacagtgg ccaagaactg ccccaatttc atcaggtttc gcctgtgcat
     4441 cctggagccc aataagcctg accacgtgac atctcagcct ctggatgagg gcttcggcgc
     4501 catcgtgaag gcctgcaaga gcctgaggcg cctgtctctg agcggcctgc tgaccgacca
     4561 ggtgttcctg tacatcggca tgtatgccaa ccagctggag atgctgtcta tcgcctttgc
     4621 cggcgacaca gataagggca tgctgtacgt gctgaatggc tgtaagaaga tgaagaagct
     4681 ggagatcaga gacagccctt ttggcgatac cgccctgctg gcagacgtga gcaagtatga
     4741 gacaatgcgg agcctgtgga tgagctcctg cgaggtgacc ctgagcggct gtaagagact
     4801 ggccgagaag gccccatggc tgaacgtgga gatcatcaac gagaatgaca acaataggat
     4861 ggaggagaat ggccacgagg gccgccagaa ggtggataag ctgtatctgt ataggaccgt
     4921 cgtcgggact cggatggatg cccccccctt tgtctggatt ctgtgagcgg caagcttatc
     4981 gataccgtcg acctcgagtc atgtaattag ttatgtcacg cttacattca cgccctcccc
     5041 ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt
     5101 atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     5161 ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt
     5221 tgggacgctc gaaggcttta atttgctggc caccgaagtc ggtgatgctg tcgccgaaga
     5281 agttaagaaa atccttgctt aaaaagattc tcttttttta tgatatttgt acataaactt
     5341 tataaatgaa attcataata gaaacgacac gaaattacaa aatggaatat gttcataggg
     5401 tagacgaaac tatatacgca atctacatac atttatcaag aaggagaaaa aggaggatgt
     5461 aaaggaatac aggtaagcaa attgatacta atggctcaac gtgataagga aaaagaattg
     5521 cactttaaca ttaatattga caaggaggag ggcaccacac aaaaagttag gtgtaacaga
     5581 aaatcatgaa actatgattc ctaatttata tattggagga ttttctctaa aaaaaaaaaa
     5641 atacaacaaa taaaaaacac tcaatgacct gaccatttga tggagtttaa gtcaatacct
     5701 ccttgaacca tttcccataa tggtgaaagt tccctcaaga attttactct gtcagaaacg
     5761 gccttaacga cgtaggccgg ccatttaaat gtttaaacgg ccacgctggc cccctatagt
     5821 gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     5881 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     5941 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc
     6001 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     6061 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     6121 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     6181 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     6241 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     6301 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     6361 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     6421 attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     6481 gtgcggtatt tcacaccgca tagggtaata actgatataa ttaaattgaa gctctaattt
     6541 gtgagtttag tattaggatc tctacccttg gcgaaaagtc ctctgccaac aatgatgata
     6601 tctgatccac cacttacaac ttcgtcgacg gttctgtact gctgacccaa tatgcgtata
     6661 tataccaatc taagtctgtg ctccttcctt cgttcttcct tctgttcgga gattaccgaa
     6721 tcaaaaaaat ttcaaggaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg
     6781 aaaaggtggt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     6841 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     6901 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     6961 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     7021 tgataataat ggtttcttag acgtcggcgc gccgaggacg tcaggtggca cttttcgggg
     7081 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     7141 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     7201 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     7261 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     7321 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     7381 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     7441 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     7501 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     7561 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     7621 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     7681 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     7741 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     7801 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     7861 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     7921 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     7981 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     8041 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     8101 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     8161 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     8221 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     8281 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     8341 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     8401 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     8461 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     8521 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     8581 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     8641 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     8701 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     8761 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     8821 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     8881 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     8941 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     9001 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     9061 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     9121 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     9181 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tcggaattaa
     9241 ccctcactaa agggaacaaa agctg
//
