LOCUS       Exported                9272 bp ds-DNA     circular SYN 20-MAY-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9272)
  AUTHORS   Sebastian Schuck
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Sep 29, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9272
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    36..696
                     /label=S.cerevisiae LEU2 upstream sequence
                     /note="S.cerevisiae LEU2 upstream sequence"
     misc_feature    697..2496
                     /label=C.glabrata LEU2 gene
                     /note="C.glabrata LEU2 gene"
     promoter        2509..3213
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             3239..4963
                     /codon_start=1
                     /label=AtAFB2
                     /note="AFB2"
                     /translation="MNYFPDEVIEHVFDFVTSHKDRNAISLVCKSWYKIERYSRQKVFI
                     GNCYAINPERLLRRFPCLKSLTLKGKPHFADFNLVPHEWGGFVLPWIEALARSRVGLEE
                     LRLKRMVVTDESLELLSRSFVNFKSLVLVSCEGFTTDGLASIAANCRHLRDLDLQENEI
                     DDHRGQWLSCFPDTCTTLVTLNFACLEGETNLVALERLVARSPNLKSLKLNRAVPLDAL
                     ARLMACAPQIVDLGVGSYENDPDSESYLKLMAVIKKCTSLRSLSGFLEAAPHCLSAFHP
                     ICHNLTSLNLSYAAEIHGSHLIKLIQHCKKLQRLWILDSIGDKGLEVVASTCKELQELR
                     VFPSDLLGGGNTAVTEEGLVAISAGCPKLHSILYFCQQMTNAALVTVAKNCPNFIRFRL
                     CILEPNKPDHVTSQPLDEGFGAIVKACKSLRRLSLSGLLTDQVFLYIGMYANQLEMLSI
                     AFAGDTDKGMLYVLNGCKKMKKLEIRDSPFGDTALLADVSKYETMRSLWMSSCEVTLSG
                     CKRLAEKAPWLNVEIINENDNNRMEENGHEGRQKVDKLYLYRTVVGTRMDAPPFVWIL"
     primer_bind     complement(4980..4996)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4999..5246
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             5251..5774
                     /label=S.cerevisiae LEU2 downstream sequence
                     /note="S.cerevisiae LEU2 downstream sequence"
     promoter        complement(5820..5838)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5845..5861)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6002..6457
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        7095..7199
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7200..8060
                     /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"
     rep_origin      8231..8819
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    9107..9128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9143..9173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9181..9197
                     /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     9205..9221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9242..9260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggccggccat ttaaatgttt aaacggccac gctggccgag gagaacttct agtatatcta
       61 catacctaat attattgcct tattaaaaat ggaatcccaa caattacatc 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 gccccacttc 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 tcttgaacca tttcccataa tggtgaaagt tccctcaaga attttactct gtcagaaacg
     5761 gccttaacga cgtaggccgg ccatttaaat gtttaaacgg ccacgctggc ccaattcgcc
     5821 ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     5881 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     5941 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     6001 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     6061 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     6241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6301 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     6361 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     6421 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     6481 cgcatctgtg cggtatttca caccgcatag ggtaataact gatataatta aattgaagct
     6541 ctaatttgtg agtttagtat taggatctct acccttggcg aaaagtcctc tgccaacaat
     6601 gatgatatct gatccaccac ttacaacttc gtcgacggtt ctgtactgct gacccaatat
     6661 gcgtatatat accaatctaa gtctgtgctc cttccttcgt tcttccttct gttcggagat
     6721 taccgaatca aaaaaatttc aaggaaaccg aaatcaaaaa aaagaataaa aaaaaaatga
     6781 tgaattgaaa aggtggtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     6841 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     6901 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     6961 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     7021 aatgtcatga taataatggt ttcttagacg tcggcgcgcc gaggacgtca ggtggcactt
     7081 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     7141 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     7201 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     7261 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     7321 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     7381 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     7441 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     7501 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     7561 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     7621 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     7681 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     7741 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     7801 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     7861 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     7921 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     7981 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     8041 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     8101 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     8161 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     8221 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     8281 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     8341 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     8401 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     8461 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     8521 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     8581 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     8641 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     8701 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     8761 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     8821 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     8881 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     8941 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     9001 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     9061 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     9121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcctatg ttgtgtggaa
     9181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg
     9241 gaattaaccc tcactaaagg gaacaaaagc tg
//
