LOCUS       Exported                7987 bp DNA     circular SYN 10-SEP-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7987)
  AUTHORS   11111111
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        184..888
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             934..954
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             970..1311
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     promoter        1317..1335
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1354..1380
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      1828..2015
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for alcohol dehydrogenase 
                     1"
     CDS             complement(2132..3226)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIEATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(3227..3632)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(3674..3690)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3698..3714
                     /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(3722..3752)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3767..3788
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    complement(3843..3876)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    complement(3954..3987)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     rep_origin      complement(4227..4815)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4986..5846)
                     /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"
     promoter        complement(5847..5951)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6233..7397
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tgtctgtttg agtacgcttt caattcattt gggtgtgcac tttattatgt tacaatatgg
       61 aagggaactt tacacttctc ctatgcacat atattaatta aagtccaatg ctagtagaga
      121 aggggggtaa cacccctccg cgctcttttc cgattttttt ctaaaccgtg gaatatttcg
      181 gatatccttt tgttgtttcc gggtgtacaa tatggacttc ctcttttctg gcaaccaaac
      241 ccatacatcg ggattcctat aataccttcg ttggtctccc taacatgtag gtggcggagg
      301 ggagatatac aatagaacag ataccagaca agacataatg ggctaaacaa gactacacca
      361 attacactgc ctcattgatg gtggtacata acgaactaat actgtagccc tagacttgat
      421 agccatcatc atatcgaagt ttcactaccc tttttccatt tgccatctat tgaagtaata
      481 ataggcgcat gcaacttctt ttcttttttt ttcttttctc tctcccccgt tgttgtctca
      541 ccatatccgc aatgacaaaa aaatgatgga agacactaaa ggaaaaaatt aacgacaaag
      601 acagcaccaa cagatgtcgt tgttccagag ctgatgaggg gtatctcgaa gcacacgaaa
      661 ctttttcctt ccttcattca cgcacactac tctctaatga gcaacggtat acggccttcc
      721 ttccagttac ttgaatttga aataaaaaaa agtttgctgt cttgctatca agtataaata
      781 gacctgcaat tattaatctt ttgtttcctc gtcattgttc tcgttccctt tcttccttgt
      841 ttctttttct gcacaatatt tcaagctata ccaagcatac aatcaactcc aagctttgca
      901 aagatggata aagcggaatt aattcccgag cctccaaaaa agaagagaaa ggtcgaattg
      961 ggtaccgccg ccaattttaa tcaaagtggg aatattgctg atagctcatt gtccttcact
     1021 ttcactaaca gtagcaacgg tccgaacctc ataacaactc aaacaaattc tcaagcgctt
     1081 tcacaaccaa ttgcctcctc taacgttcat gataacttca tgaataatga aatcacggct
     1141 agtaaaattg atgatggtaa taattcaaaa ccactgtcac ctggttggac ggaccaaact
     1201 gcgtataacg cgtttggaat cactacaggg atgtttaata ccactacaat ggatgatgta
     1261 tataactatc tattcgatga tgaagatacc ccaccaaacc caaaaaaaga gatctttaat
     1321 acgactcact atagggcgag cgccgccatg gagtacccat acgacgtacc agattacgct
     1381 catatggcca tggaggccag tgaattccac ccgggtgggc atcgatacgg gatccatcga
     1441 gctcgagctg cagatgaatc gtagatactg aaaaaccccg caagttcact tcaactgtgc
     1501 atcgtgcacc atctcaattt ctttcattta tacatcgttt tgccttcttt tatgtaacta
     1561 tactcctcta agtttcaatc ttggccatgt aacctctgat ctatagaatt ttttaaatga
     1621 ctagaattaa tgcccatctt ttttttggac ctaaattctt catgaaaata tattacgagg
     1681 gcttattcag aagctttgga cttcttcgcc agaggtttgg tcaagtctcc aatcaaggtt
     1741 gtcggcttgt ctaccttgcc agaaatttac gaaaagatgg aaaagggtca aatcgttggt
     1801 agatacgttg ttgacacttc taaataagcg aatttcttat gatttatgat ttttattatt
     1861 aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa
     1921 cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag
     1981 catgaggtcg ctcttattga ccacacctct accggccggt cgaaattccc ctaccctatg
     2041 aacatattcc attttgtaat ttcgtgtcgt ttctattatg aatttcattt ataaagttta
     2101 tgtacaaata tcataaaaaa agagaatctt tttaagcaag gattttctta acttcttcgg
     2161 cgacagcatc accgacttcg gtggtactgt tggaaccacc taaatcacca gttctgatac
     2221 ctgcatccaa aaccttttta actgcatctt caatggcctt accttcttca ggcaagttca
     2281 atgacaattt caacatcatt gcagcagaca agatagtggc gatagggttg accttattct
     2341 ttggcaaatc tggagcagaa ccgtggcatg gttcgtacaa accaaatgcg gtgttcttgt
     2401 ctggcaaaga ggccaaggac gcagatggca acaaacccaa ggaacctggg ataacggagg
     2461 cttcatcgga gatgatatca ccaaacatgt tgctggtgat tataatacca tttaggtggg
     2521 ttgggttctt aactaggatc atggcggcag aatcaatcaa ttgatgttga accttcaatg
     2581 taggaaattc gttcttgatg gtttcctcca cagtttttct ccataatctt gaagaggcca
     2641 aaacattagc tttatccaag gaccaaatag gcaatggtgg ctcatgttgt agggccatga
     2701 aagcggccat tcttgtgatt ctttgcactt ctggaacggt gtattgttca ctatcccaag
     2761 cgacaccatc accatcgtct tcctttctct taccaaagta aatacctccc actaattctc
     2821 tgacaacaac gaagtcagta cctttagcaa attgtggctt gattggagat aagtctaaaa
     2881 gagagtcgga tgcaaagtta catggtctta agttggcgta caattgaagt tctttacgga
     2941 tttttagtaa accttgttca ggtctaacac tacctgtacc ccatttagga ccacccacag
     3001 cacctaacaa aacggcatca gccttcttgg aggcttccag cgcctcatct ggaagtggga
     3061 cacctgtagc ttcgatagca gcaccaccaa ttaaatgatt ttcgaaatcg aacttgacat
     3121 tggaacgaac atcagaaata gctttaagaa ccttaatggc ttcggctgtg atttcttgac
     3181 caacgtggtc acctggcaaa acgacgatct tcttaggggc agacattaga atggtatatc
     3241 cttgaaatat atatatatat tgctgaaatg taaaaggtaa gaaaagttag aaagtaagac
     3301 gattgctaac cacctattgg aaaaaacaat aggtccttaa ataatattgt caacttcaag
     3361 tattgtgatg caagcattta gtcatgaacg cttctctatt ctatatgaaa agccggttcc
     3421 ggcgctctca cctttccttt ttctcccaat ttttcagttg aaaaaggtat atgcgtcagg
     3481 cgacctctga aattaacaaa aaatttccag tcatcgaatt tgattctgtg cgatagcgcc
     3541 cctgtgtgtt ctcgttatgt tgaggaaaaa aataatggtt gctaagagat tcgaactctt
     3601 gcatcttacg atacctgagt attcccacag ttgggggatc tcgactctag ctagaggatc
     3661 aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3721 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3781 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3841 tgataacttc gtataatgta tgctatacga agttattagg tctgaagagg agtttacgtc
     3901 cagccaagct agcttggctg caggtcgagc ggccgcgatc cggaaccctt aatataactt
     3961 cgtataatgt atgctatacg aagttatcag ctgcattaat gaatcggcca acgcgcgggg
     4021 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4081 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4141 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4201 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4261 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4321 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4381 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4441 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4501 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4561 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4621 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4681 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4741 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4801 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4861 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4921 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4981 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5041 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5101 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5161 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5221 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5281 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5341 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5401 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5461 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5521 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5581 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5641 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5701 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5761 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5821 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5881 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5941 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     6001 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     6061 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     6121 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     6181 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca taacgcattt
     6241 aagcataaac acgcactatg ccgttcttct catgtatata tatatacagg caacacgcag
     6301 atataggtgc gacgtgaaca gtgagctgta tgtgcgcagc tcgcgttgca ttttcggaag
     6361 cgctcgtttt cggaaacgct ttgaagttcc tattccgaag ttcctattct ctagctagaa
     6421 agtataggaa cttcagagcg cttttgaaaa ccaaaagcgc tctgaagacg cactttcaaa
     6481 aaaccaaaaa cgcaccggac tgtaacgagc tactaaaata ttgcgaatac cgcttccaca
     6541 aacattgctc aaaagtatct ctttgctata tatctctgtg ctatatccct atataaccta
     6601 cccatccacc tttcgctcct tgaacttgca tctaaactcg acctctacat tttttatgtt
     6661 tatctctagt attactcttt agacaaaaaa attgtagtaa gaactattca tagagtgaat
     6721 cgaaaacaat acgaaaatgt aaacatttcc tatacgtagt atatagagac aaaatagaag
     6781 aaaccgttca taattttctg accaatgaag aatcatcaac gctatcactt tctgttcaca
     6841 aagtatgcgc aatccacatc ggtatagaat ataatcgggg atgcctttat cttgaaaaaa
     6901 tgcacccgca gcttcgctag taatcagtaa acgcgggaag tggagtcagg ctttttttat
     6961 ggaagagaaa atagacacca aagtagcctt cttctaacct taacggacct acagtgcaaa
     7021 aagttatcaa gagactgcat tatagagcgc acaaaggaga aaaaaagtaa tctaagatgc
     7081 tttgttagaa aaatagcgct ctcgggatgc atttttgtag aacaaaaaag aagtatagat
     7141 tctttgttgg taaaatagcg ctctcgcgtt gcatttctgt tctgtaaaaa tgcagctcag
     7201 attctttgtt tgaaaaatta gcgctctcgc gttgcatttt tgttttacaa aaatgaagca
     7261 cagattcttc gttggtaaaa tagcgctttc gcgttgcatt tctgttctgt aaaaatgcag
     7321 ctcagattct ttgtttgaaa aattagcgct ctcgcgttgc atttttgttc tacaaaatga
     7381 agcacagatg cttcgttgct tgcatgcctg caggtcgaga tccgggatcg aagaaatgat
     7441 ggtaaatgaa ataggaaatc aaggagcatg aaggcaaaag acaaatataa gggtcgaacg
     7501 aaaaataaag tgaaaagtgt tgatatgatg tatttggctt tgcggcgccg aaaaaacgag
     7561 tttacgcaat tgcacaatca tgctgactct gtggcggacc cgcgctcttg ccggcccggc
     7621 gataacgctg ggcgtgaggc tgtgcccggc ggagtttttt gcgcctgcat tttccaaggt
     7681 ttaccctgcg ctaaggggcg agattggaga agcaataaga atgccggttg gggttgcgat
     7741 gatgacgacc acgacaactg gtgtcattat ttaagttgcc gaaagaacct gagtgcattt
     7801 gcaacatgag tatactagaa gaatgagcca agacttgcga gacgcgagtt tgccggtggt
     7861 gcgaacaata gagcgaccat gaccttgaag gtgagacgcg cataaccgct agagtacttt
     7921 gaagaggaaa cagcaatagg gttgctacca gtataaatag acaggtacat acaacactgg
     7981 aaatggt
//
