LOCUS       Exported                8628 bp ds-DNA     circular SYN 01-JUL-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8628)
  AUTHORS   Bruegger Group
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Sep 29, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8628
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        311..498
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     misc_feature    499..706
                     /label=HIS3''
     terminator      complement(724..971)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(979..2007)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(2008..2351)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     misc_feature    2430..3153
                     /label=shortened ADH promoter
     CDS             3186..4913
                     /codon_start=1
                     /label=OsTIR1(F74G)
                     /translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV
                     GNCYAVRAGRVAARFPNVRALTVKGKPHGADFNLVPPDWGGYAGPWIEAAARGCHGLEE
                     LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV
                     EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL
                     AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP
                     LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR
                     VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL
                     CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI
                     AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG
                     CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL"
     misc_feature    4920..5171
                     /label=CYC terminator
                     /note="CYC terminator"
     misc_feature    5185..5432
                     /label=HIS3'
     rep_origin      complement(5693..6148)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6316..6334
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6374..6392)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    6437..6453
                     /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(6461..6491)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(6815..7403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7574..8434)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8435..8539)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
       61 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
      121 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
      181 taattccgtt ttaagagctt ggtgagcgct aggagtcact gccaggtatc gtttgaacac
      241 ggcattagtc agggaagtca taacacagtc ctttcccgca attttctttt tctattactc
      301 ttggcctcct ctagtacact ctatattttt ttatgcctcg gtaatgattt tcattttttt
      361 ttttccacct agcggatgac tctttttttt tcttagcgat tggcattatc acataatgaa
      421 ttatacatta tataaagtaa tgtgatttct tcgaagaata tactaaaaaa tgagcaggca
      481 agataaacga aggcaaagat gacagagcag aaagccctag taaagcgtat tacaaatgaa
      541 accaagattc agattgcgat ctctttaaag ggtggtcccc tagcgataga gcactcgatc
      601 ttcccagaaa aagaggcaga agcagtagca gaacaggcca cacaatcgca agtgattaac
      661 gtccacacag gtatagggtt tctggaccat atgatacatg ctctggatca tcgatgaatt
      721 cgagctcgtt aaagccttcg agcgtcccaa aaccttctca agcaaggttt tcagtataat
      781 gttacatgcg tacacgcgtc tgtacagaaa aaaaagaaaa atttgaaata taaataacgt
      841 tcttaatact aacataacta taaaaaaata aatagggacc tagacttcag gttgtctaac
      901 tccttccttt tcggttagag cggatgtggg gggagggcgt gaatgtaagc gtgacataac
      961 taattacatg actcgagatt attcctttgc cctcggacga gtgctggggc gtcggtttcc
     1021 actatcggcg agtacttcta cacagccatc ggtccagacg gccgcgcttc tgcgggcgat
     1081 ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc
     1141 ccaagctgca tcatcgaaat tgccgtcaac caagctctga tagagttggt caagaccaat
     1201 gcggagcata tacgcccgga gccgcggcga tcctgcaagc tccggatgcc tccgctcgaa
     1261 gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc cagaagaaga tgttggcgac
     1321 ctcgtattgg gaatccccga acatcgcctc gctccagtca atgaccgctg ttatgcggcc
     1381 attgtccgtc aggacattgt tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg
     1441 gcagtcctcg gcccaaagca tcagctcatc gagagcctgc gcgacggacg cactgacggt
     1501 gtcgtccatc acagtttgcc agtgatacac atggggatca gcaatcgcgc atatgaaatc
     1561 acgccatgta gtgtattgac cgattccttg cggtccgaat gggccgaacc cgctcgtctg
     1621 gctaagatcg gccgcagcga tcgcatccat ggcctccgcg accggctgca gaacagcggg
     1681 cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt gcacggcggg agatgcaata
     1741 ggtcaggctc tcgctgaatt ccccaatgtc aagcacttcc ggaatcggga gcgcggccga
     1801 tgcaaagtgc cgataaacat aacgatcttt gtagaaacca tcggcgcagc tatttacccg
     1861 caggacatat ccacgccctc ctacatcgaa gctgaaagca cgagattctt cgccctccga
     1921 gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc agaaacttct cgacagacgt
     1981 cgcggtgagt tcaggctttt tacccatggt tgtttatgtt cggatgtgat gtgagaactg
     2041 tatcctagca agattttaaa aggaagtata tgaaagaaga acctcagtgg caaatcctaa
     2101 ccttttatat ttctctacag gggcgcggcg tggggacaat tcaacgcgtc tgtgagggga
     2161 gcgtttccct gctcgcaggt ctgcagcgag gagccgtaat ttttgcttcg cgccgtgcgg
     2221 ccatcaaaat gtatggatgc aaatgattat acatggggat gtatgggcta aatgtacggg
     2281 cgacagtcac atcatgcccc tgagctgcgc acgtcaagac tgtcaaggag ggtattctgg
     2341 gcctccatgt cgctggccgg gtgacccggc ggggacgagg caagctaaac agatctggcg
     2401 cgccttaatt aacccgggga tccgagctca tccttttgtt gtttccgggt gtacaatatg
     2461 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
     2521 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
     2581 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga
     2641 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt
     2701 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct
     2761 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac
     2821 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga
     2881 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc
     2941 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt
     3001 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
     3061 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
     3121 gcatacaatc aactccaagc tggccgctct agaagaactt agtttcgacg gattctagaa
     3181 ctagtatgac ttattttcct gaagaagttg tagaacacat attttccttt ttacctgctc
     3241 aaagagatag aaacaccgtc tccttggtct gtaaggtttg gtacgaaatc gaaagattgt
     3301 ctagaagagg tgtttttgtt ggtaattgtt acgcagttag agctggtaga gttgctgcaa
     3361 gattcccaaa cgttagagca ttgactgtta aaggtaaacc tcatggtgct gatttcaatt
     3421 tggttccacc tgattggggt ggttatgctg gtccatggat tgaagctgca gctagaggtt
     3481 gtcacggttt ggaagaattg agaatgaaga gaatggttgt ttctgatgaa tcattggaat
     3541 tgttagcaag atcttttcct agattcagag ctttggtttt gatctcatgt gaaggttttt
     3601 ccaccgatgg tttagcagct gttgcttcac attgtaagtt gttgagagaa ttggatttgc
     3661 aagaaaacga agttgaagat agaggtccaa gatggttatc ttgttttcct gattcctgta
     3721 caagtttggt ttctttgaac ttcgcatgta tcaagggtga agttaacgct ggtagtttgg
     3781 aaagattagt ttctagatca ccaaatttga gatctttgag attgaacaga tccgttagtg
     3841 ttgatacctt ggcaaagatc ttgttgagaa ctcctaattt ggaagatttg ggtactggta
     3901 acttgacaga tgatttccaa acagaatcat acttcaagtt gacctccgct ttggaaaagt
     3961 gtaagatgtt gagatctttg tcaggttttt gggatgcaag tccagtttgt ttgtctttta
     4021 tatatccttt gtgtgctcaa ttgactggtt tgaatttgtc ttacgcacca actttggatg
     4081 ctagtgattt gacaaagatg atctctagat gtgttaagtt gcaaagattg tgggttttgg
     4141 attgtatctc agataaaggt ttgcaagttg ttgcttcttc ttgtaaggat ttgcaagaat
     4201 tgagagtttt tccaagtgat ttctatgttg caggttactc tgctgttaca gaagaaggtt
     4261 tagttgcagt ttcattgggt tgtcctaagt tgaactcctt gttgtacttc tgtcatcaaa
     4321 tgaccaacgc agctttggtt actgttgcta aaaattgtcc aaacttcact agattcagat
     4381 tgtgtatctt ggaaccaggt aaacctgatg ttgttacatc acaaccttta gatgaaggtt
     4441 ttggtgcaat cgttagagag tgtaagggtt tgcaaagatt gtccatcagt ggtttgttga
     4501 cagataaggt ttttatgtac atcggtaaat acgctaagca attggaaatg ttgtcaatcg
     4561 cattcgctgg tgactccgat aagggtatga tgcatgttat gaacggttgt aagaacttaa
     4621 gaaagttgga aatcagagat tctccatttg gtgacgcagc tttgttgggt aatttcgcaa
     4681 gatatgaaac aatgagatct ttgtggatgt ccagttgtaa cgttaccttg aaaggttgtc
     4741 aagttttagc ttcaaagatg ccaatgttga acgttgaagt tattaacgaa agagatggtt
     4801 ccaatgaaat ggaagaaaac catggtgact tgcctaaagt tgaaaagtta tacgtctaca
     4861 gaacaaccgc aggtgccaga gatgatgctc ctaacttcgt caagatttta tgactcgagt
     4921 catgtaatta gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga
     4981 aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt
     5041 agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta
     5101 cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt
     5161 aatttgcggc cggtcgacct gcagcgtacg cagttgtcga acttggtttg caaagggaga
     5221 aagtaggaga tctctcttgc gagatgatcc cgcattttct tgaaagcttt gcagaggcta
     5281 gcagaattac cctccacgtt gattgtctgc gaggcaagaa tgatcatcac cgtagtgaga
     5341 gtgcgttcaa ggctcttgcg gttgccataa gagaagccac ctcgcccaat ggtaccaacg
     5401 atgttccctc caccaaaggt gttcttatgt agtgacaccg attatttaaa gctgcagcat
     5461 acgatatata tacatgtgta tatatgtata cctatgaatg tcagtaagta tgtatacgaa
     5521 cagtatgata ctgaagatga caaggtaatg catcattcta tacgtgtcat tctgaacgag
     5581 gcgcgctttc cttttttctt tttgcttttt cttttttttt ctcttgaact cgacggatca
     5641 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta
     5701 aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     5761 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     5821 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     5881 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     5941 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     6001 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     6061 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     6121 gcgcttaatg cgccgctaca gggcgcgtcg cgccattcgc cattcaggct gcgcaactgt
     6181 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa ggggggatgt
     6241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     6301 acggccagtg aattgtaata cgactcacta tagggcgaat tggagggggg ggcccggtac
     6361 ccagcttttg ttccctttag tgagggttaa ttccgagctt ggcgtaatca tggtcatagc
     6421 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatagga gccggaagca
     6481 taaagtgtaa agcctggggt gcctaatgag tgaggtaact cacattaatt gcgttgcgct
     6541 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6601 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6661 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6721 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6781 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctcggc ccccctgacg
     6841 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6901 accaggcgtt cccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6961 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     7021 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7081 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7141 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7201 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     7261 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7321 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7381 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7441 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7501 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7561 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7621 ttcgttcatc catagttgcc tgactgcccg tcgtgtagat aactacgata cgggagggct
     7681 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7741 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7801 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7861 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7921 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7981 tgtgaaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8041 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8101 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8161 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8221 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8281 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8341 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8401 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8461 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8521 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc ttattatcat
     8581 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgc
//
