LOCUS       Exported                8245 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 8245)
  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..8245
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        311..498
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     misc_feature    499..706
                     /label=HIS3''
                     /note="HIS3''"
     terminator      complement(854..1041)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     misc_feature    1052..1624
                     /label=natR
                     /note="natR"
     promoter        complement(1625..1968)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     misc_feature    2047..2770
                     /label=ADH1 prom short
     CDS             2803..4530
                     /codon_start=1
                     /label=Os TIR1 (F74G)
                     /translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV
                     GNCYAVRAGRVAARFPNVRALTVKGKPHGADFNLVPPDWGGYAGPWIEAAARGCHGLEE
                     LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV
                     EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL
                     AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP
                     LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR
                     VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL
                     CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI
                     AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG
                     CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL"
     misc_feature    4537..4788
                     /label=CYC terminator
                     /note="CYC terminator"
     misc_feature    4802..5049
                     /label=HIS3'
                     /note="HIS3'"
     rep_origin      complement(5310..5765)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5933..5951
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5991..6009)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6030..6046)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6078..6108)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(6432..7020)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7191..8051)
                     /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(8052..8156)
                     /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 cgagctcgat tacaacaggt gttgtcctct gaggacataa aatacacacc gagattcatc
      781 aactcattgc tggagttagc atatctacaa ttgggtgaaa tggggagcga tttgcaggca
      841 tttgctcggc atgccggtag aggtgtggtc aataagagcg acctcatgct atacctgaga
      901 aagcaacctg acctacagga aagagttact caagaacaag aattttcgtt ttaaaaccta
      961 agagtcactt taaaatttgt atacacttat tttttttata acttatttaa taataaaaat
     1021 cataaatcat aagaaattcg cctcgagatt aggggcaggg catgctcatg tagagcgcct
     1081 gccgctcgcc gtccgaggcg gtgccgtcgt acagggcggt gtccaggccg cagagggtga
     1141 accccatccg ccggtacgcg tggatcgccg gtgcgttgac gttggtgacc tccagccaga
     1201 ggtgcccggc gccccgctcg ccggcgaact ccgtcgcgag ccccatcaac gcgcgcccga
     1261 ccccgtgccc ccggtgctcc ggggcgacct cgatgtcctc gacggtcagc cggcggttcc
     1321 acgccgagta cgagatgacc acgaagcccg ccaggtcgcc gtcgtccccg tacgcgacga
     1381 acgtccggga gtccgggtcg ccgtcctccc cgtcgtccga ttcgtcgtcc gattcgtcgt
     1441 cggggaacac cttggtcagg ggcgggtcca ccggcacctc ccgcagggtg aagccgtccc
     1501 cggtggcggt gacgcggaag acggtgtcgg tggtgaagga cccatccagt gcctcgatgg
     1561 cctcggcgtc ccccgggaca ctggtgcggt accggtaagc cgtgtcgtca agagtggtac
     1621 ccatggttgt ttatgttcgg atgtgatgtg agaactgtat cctagcaaga ttttaaaagg
     1681 aagtatatga aagaagaacc tcagtggcaa atcctaacct tttatatttc tctacagggg
     1741 cgcggcgtgg ggacaattca acgcgtctgt gaggggagcg tttccctgct cgcaggtctg
     1801 cagcgaggag ccgtaatttt tgcttcgcgc cgtgcggcca tcaaaatgta tggatgcaaa
     1861 tgattataca tggggatgta tgggctaaat gtacgggcga cagtcacatc atgcccctga
     1921 gctgcgcacg tcaagactgt caaggagggt attctgggcc tccatgtcgc tggccgggtg
     1981 acccggcggg gacgaggcaa gctaaacaga tctggcgcgc cttaattaac ccggggatcc
     2041 gagctcatcc ttttgttgtt tccgggtgta caatatggac ttcctctttt ctggcaacca
     2101 aacccataca tcgggattcc tataatacct tcgttggtct ccctaacatg taggtggcgg
     2161 aggggagata tacaatagaa cagataccag acaagacata atgggctaaa caagactaca
     2221 ccaattacac tgcctcattg atggtggtac ataacgaact aatactgtag ccctagactt
     2281 gatagccatc atcatatcga agtttcacta ccctttttcc atttgccatc tattgaagta
     2341 ataataggcg catgcaactt cttttctttt tttttctttt ctctctcccc cgttgttgtc
     2401 tcaccatatc cgcaatgaca aaaaaatgat ggaagacact aaaggaaaaa attaacgaca
     2461 aagacagcac caacagatgt cgttgttcca gagctgatga ggggtatctc gaagcacacg
     2521 aaactttttc cttccttcat tcacgcacac tactctctaa tgagcaacgg tatacggcct
     2581 tccttccagt tacttgaatt tgaaataaaa aaaagtttgc tgtcttgcta tcaagtataa
     2641 atagacctgc aattattaat cttttgtttc ctcgtcattg ttctcgttcc ctttcttcct
     2701 tgtttctttt tctgcacaat atttcaagct ataccaagca tacaatcaac tccaagctgg
     2761 ccgctctaga agaacttagt ttcgacggat tctagaacta gtatgactta ttttcctgaa
     2821 gaagttgtag aacacatatt ttccttttta cctgctcaaa gagatagaaa caccgtctcc
     2881 ttggtctgta aggtttggta cgaaatcgaa agattgtcta gaagaggtgt ttttgttggt
     2941 aattgttacg cagttagagc tggtagagtt gctgcaagat tcccaaacgt tagagcattg
     3001 actgttaaag gtaaacctca tggtgctgat ttcaatttgg ttccacctga ttggggtggt
     3061 tatgctggtc catggattga agctgcagct agaggttgtc acggtttgga agaattgaga
     3121 atgaagagaa tggttgtttc tgatgaatca ttggaattgt tagcaagatc ttttcctaga
     3181 ttcagagctt tggttttgat ctcatgtgaa ggtttttcca ccgatggttt agcagctgtt
     3241 gcttcacatt gtaagttgtt gagagaattg gatttgcaag aaaacgaagt tgaagataga
     3301 ggtccaagat ggttatcttg ttttcctgat tcctgtacaa gtttggtttc tttgaacttc
     3361 gcatgtatca agggtgaagt taacgctggt agtttggaaa gattagtttc tagatcacca
     3421 aatttgagat ctttgagatt gaacagatcc gttagtgttg ataccttggc aaagatcttg
     3481 ttgagaactc ctaatttgga agatttgggt actggtaact tgacagatga tttccaaaca
     3541 gaatcatact tcaagttgac ctccgctttg gaaaagtgta agatgttgag atctttgtca
     3601 ggtttttggg atgcaagtcc agtttgtttg tcttttatat atcctttgtg tgctcaattg
     3661 actggtttga atttgtctta cgcaccaact ttggatgcta gtgatttgac aaagatgatc
     3721 tctagatgtg ttaagttgca aagattgtgg gttttggatt gtatctcaga taaaggtttg
     3781 caagttgttg cttcttcttg taaggatttg caagaattga gagtttttcc aagtgatttc
     3841 tatgttgcag gttactctgc tgttacagaa gaaggtttag ttgcagtttc attgggttgt
     3901 cctaagttga actccttgtt gtacttctgt catcaaatga ccaacgcagc tttggttact
     3961 gttgctaaaa attgtccaaa cttcactaga ttcagattgt gtatcttgga accaggtaaa
     4021 cctgatgttg ttacatcaca acctttagat gaaggttttg gtgcaatcgt tagagagtgt
     4081 aagggtttgc aaagattgtc catcagtggt ttgttgacag ataaggtttt tatgtacatc
     4141 ggtaaatacg ctaagcaatt ggaaatgttg tcaatcgcat tcgctggtga ctccgataag
     4201 ggtatgatgc atgttatgaa cggttgtaag aacttaagaa agttggaaat cagagattct
     4261 ccatttggtg acgcagcttt gttgggtaat ttcgcaagat atgaaacaat gagatctttg
     4321 tggatgtcca gttgtaacgt taccttgaaa ggttgtcaag ttttagcttc aaagatgcca
     4381 atgttgaacg ttgaagttat taacgaaaga gatggttcca atgaaatgga agaaaaccat
     4441 ggtgacttgc ctaaagttga aaagttatac gtctacagaa caaccgcagg tgccagagat
     4501 gatgctccta acttcgtcaa gattttatga ctcgagtcat gtaattagtt atgtcacgct
     4561 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
     4621 aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat
     4681 ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa
     4741 ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcggccgg tcgacctgca
     4801 gcgtacgcag ttgtcgaact tggtttgcaa agggagaaag taggagatct ctcttgcgag
     4861 atgatcccgc attttcttga aagctttgca gaggctagca gaattaccct ccacgttgat
     4921 tgtctgcgag gcaagaatga tcatcaccgt agtgagagtg cgttcaaggc tcttgcggtt
     4981 gccataagag aagccacctc gcccaatggt accaacgatg ttccctccac caaaggtgtt
     5041 cttatgtagt gacaccgatt atttaaagct gcagcatacg atatatatac atgtgtatat
     5101 atgtatacct atgaatgtca gtaagtatgt atacgaacag tatgatactg aagatgacaa
     5161 ggtaatgcat cattctatac gtgtcattct gaacgaggcg cgctttcctt ttttcttttt
     5221 gctttttctt tttttttctc ttgaactcga cggatcatat gcggtgtgaa ataccgcaca
     5281 gatgcgtaag gagaaaatac cgcatcagga aattgtaaac gttaatattt tgttaaaatt
     5341 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     5401 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     5461 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     5521 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     5581 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     5641 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     5701 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     5761 cgcgtcgcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5821 ctcttcgcta ttacgccagc tggcgaaggg gggatgtgct gcaaggcgat taagttgggt
     5881 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga
     5941 ctcactatag ggcgaattgg aggggggggc ccggtaccca gcttttgttc cctttagtga
     6001 gggttaattc cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     6061 ccgctcacaa ttccacacaa cataggagcc ggaagcataa agtgtaaagc ctggggtgcc
     6121 taatgagtga ggtaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6181 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6241 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6301 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6361 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6421 ttgctggcgt ttttccatag gctcggcccc cctgacgagc atcacaaaaa tcgacgctca
     6481 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgttccc ccctggaagc
     6541 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6601 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     6661 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6721 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6781 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6841 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     6901 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6961 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7021 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7081 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7141 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7201 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7261 ctgcccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7321 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7381 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7441 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7501 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7561 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gaaaaaaagc ggttagctcc
     7621 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7681 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7741 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7801 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7861 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7921 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7981 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8041 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8101 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8161 tttccccgaa aagtgccacc tgacgtctta ttatcatgac attaacctat aaaaataggc
     8221 gtatcacgag gccctttcgt ctcgc
//
