LOCUS       62056_11230        9973 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9973)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        771..1476
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1522..1542
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             1558..1899
                     /codon_start=1
                     /label=GAL4 activation domain
                     /note="activation domain of the GAL4 transcriptional
                     activator"
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     promoter        1905..1923
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1942..1968
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             2113..2133
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     polyA_signal    3887..4019
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     terminator      4401..4588
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(4708..5799)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(5800..6205)
                     /label=LEU2 promoter
     promoter        complement(6295..6325)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6340..6361)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     protein_bind    6416..6449
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(6800..7388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7562..8419)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8420..8524)
                     /label=AmpR promoter
     rep_origin      8806..9970
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tgcatgcctg caggtcgaga tccgggatcg aagaaatgat ggtaaatgaa ataggaaatc
       61 aaggagcatg aaggcaaaag acaaatataa gggtcgaacg aaaaataaag tgaaaagtgt
      121 tgatatgatg tatttggctt tgcggcgccg aaaaaacgag tttacgcaat tgcacaatca
      181 tgctgactct gtggcggacc cgcgctcttg ccggcccggc gataacgctg ggcgtgaggc
      241 tgtgcccggc ggagtttttt gcgcctgcat tttccaaggt ttaccctgcg ctaaggggcg
      301 agattggaga agcaataaga atgccggttg gggttgcgat gatgacgacc acgacaactg
      361 gtgtcattat ttaagttgcc gaaagaacct gagtgcattt gcaacatgag tatactagaa
      421 gaatgagcca agacttgcga gacgcgagtt tgccggtggt gcgaacaata gagcgaccat
      481 gaccttgaag gtgagacgcg cataaccgct agagtacttt gaagaggaaa cagcaatagg
      541 gttgctacca gtataaatag acaggtacat acaacactgg aaatggttgt ctgtttgagt
      601 acgctttcaa ttcatttggg tgtgcacttt attatgttac aatatggaag ggaactttac
      661 acttctccta tgcacatata ttaattaaag tccaatgcta gtagagaagg ggggtaacac
      721 ccctccgcgc tcttttccga tttttttcta aaccgtggaa tatttcggat atccttttgt
      781 tgtttccggg tgtacaatat ggacttcctc ttttctggca accaaaccca tacatcggga
      841 ttcctataat accttcgttg gtctccctaa catgtaggtg gcggagggga gatatacaat
      901 agaacagata ccagacaaga cataatgggc taaacaagac tacaccaatt acactgcctc
      961 attgatggtg gtacataacg aactaatact gtagccctag acttgatagc catcatcata
     1021 tcgaagtttc actacccttt ttccatttgc catctattga agtaataata ggcgcatgca
     1081 acttcttttc tttttttttc ttttctctct cccccgttgt tgtctcacca tatccgcaat
     1141 gacaaaaaaa atgatggaag acactaaagg aaaaaattaa cgacaaagac agcaccaaca
     1201 gatgtcgttg ttccagagct gatgaggggt atctcgaagc acacgaaact ttttccttcc
     1261 ttcattcacg cacactactc tctaatgagc aacggtatac ggccttcctt ccagttactt
     1321 gaatttgaaa taaaaaaaag tttgctgtct tgctatcaag tataaataga cctgcaatta
     1381 ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt ctttttctgc
     1441 acaatatttc aagctatacc aagcatacaa tcaactccaa gctttgcaaa gatggataaa
     1501 gcggaattaa ttcccgagcc tccaaaaaag aagagaaagg tcgaattggg taccgccgcc
     1561 aattttaatc aaagtgggaa tattgctgat agctcattgt ccttcacttt cactaacagt
     1621 agcaacggtc cgaacctcat aacaactcaa acaaattctc aagcgctttc acaaccaatt
     1681 gcctcctcta acgttcatga taacttcatg aataatgaaa tcacggctag taaaattgat
     1741 gatggtaata attcaaaacc actgtcacct ggttggacgg accaaactgc gtataacgcg
     1801 tttggaatca ctacagggat gtttaatacc actacaatgg atgatgtata taactatcta
     1861 ttcgatgatg aagatacccc accaaaccca aaaaaagaga tctttaatac gactcactat
     1921 agggcgagcg ccgccatgga gtacccatac gacgtaccag attacgctca tatggccatg
     1981 gaggccagtg aattcggaac tgatgaatgg gagcagtggt ggaatgcctt taatgaggaa
     2041 aacctgtttt gctcagaaga aatgccatct agtgatgatg aggctactgc tgactctcaa
     2101 cattctactc ctccaaaaaa gaagagaaag gtagaagacc ccaaggactt tccttcagaa
     2161 ttgctaagtt ttttgagtca tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt
     2221 tacaccacaa aggaaaaagc tgcactgcta tacaagaaaa ttatggaaaa atattctgta
     2281 acctttataa gtaggcataa cagttataat cataacatac tgttttttct tactccacac
     2341 aggcatagag tgtctgctat taataactat gctcaaaaat tgtgtacctt tagcttttta
     2401 atttgtaaag gggttaataa ggaatatttg atgtatagtg ccttgactag agatccattt
     2461 tctgttattg aggaaagttt gccaggtggg ttaaaggagc atgattttaa tccagaagaa
     2521 gcagaggaaa ctaaacaagt gtcctggaag cttgtaacag agtatgcaat ggaaacaaaa
     2581 tgtgatgatg tgttgttatt gcttgggatg tacttggaat ttcagtacag ttttgaaatg
     2641 tgtttaaaat gtattaaaaa agaacagccc agccactata agtaccatga aaagcattat
     2701 gcaaatgctg ctatatttgc tgacagcaaa aaccaaaaaa ccatatgcca acaggctgtt
     2761 gatactgttt tagctaaaaa gcgggttgat agcctacaat taactagaga acaaatgtta
     2821 acaaacagat ttaatgatct tttggatagg atggatataa tgtttggttc tacaggctct
     2881 gctgacatag aagaatggat ggctggagtt gcttggctac actgtttgtt gcccaaaatg
     2941 gattcagtgg tgtatgactt tttaaaatgc atggtgtaca acattcctaa aaaaagatac
     3001 tggctgttta aaggaccaat tgatagtggt aaaactacat tagcagctgc tttgcttgaa
     3061 ttatgtgggg ggaaagcttt aaatgttaat ttgcccttgg acaggctgaa ctttgagcta
     3121 ggagtagcta ttgaccagtt tttagtagtt tttgaggatg taaagggcac tggaggggag
     3181 tccagagatt tgccttcagg tcagggaatt aataacctgg acaatttaag ggattatttg
     3241 gatggcagtg ttaaggtaaa cttagaaaag aaacacctaa ataaaagaac tcaaatattt
     3301 ccccctggaa tagtcaccat gaatgagtac agtgtgccta aaacactgca ggccagattt
     3361 gtaaaacaaa tagattttag gcccaaagat tatttaaagc attgcctgga acgcagtgag
     3421 tttttgttag aaaagagaat aattcaaagt ggcattgctt tgcttcttat gttaatttgg
     3481 tacagacctg tggctgagtt tgctcaaagt attcagagca gaattgtgga gtggaaagag
     3541 agattggaca aagagtttag tttgtcagtg tatcaaaaaa tgaagtttaa tgtggctatg
     3601 ggaattggag ttttagattg gctaagaaac agtgatgatg atgatgaaga cagccaggaa
     3661 aatgctgata aaaatgaaga tggtggggag aagaacatgg aagactcagg gcatgaaaca
     3721 ggcattgatt cacagtccca aggctcattt caggcccctc agtcctcaca gtctgttcat
     3781 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3841 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3901 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3961 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctcgag
     4021 ctgcagatga atcgtagata ctgaaaaacc ccgcaagttc acttcaactg tgcatcgtgc
     4081 accatctcaa tttctttcat ttatacatcg ttttgccttc ttttatgtaa ctatactcct
     4141 ctaagtttca atcttggcca tgtaacctct gatctataga attttttaaa tgactagaat
     4201 taatgcccat cttttttttg gacctaaatt cttcatgaaa atatattacg agggcttatt
     4261 cagaagcttt ggacttcttc gccagaggtt tggtcaagtc tccaatcaag gttgtcggct
     4321 tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg tcaaatcgtt ggtagatacg
     4381 ttgttgacac ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag
     4441 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
     4501 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg
     4561 tcgctcttat tgaccacacc tctaccggcc ggtcgaaatt cccctaccct atgaacatat
     4621 tccattttgt aatttcgtgt cgtttctatt atgaatttca tttataaagt ttatgtacaa
     4681 atatcataaa aaaagagaat ctttttaagc aaggattttc ttaacttctt cggcgacagc
     4741 atcaccgact tcggtggtac tgttggaacc acctaaatca ccagttctga tacctgcatc
     4801 caaaaccttt ttaactgcat cttcaatggc cttaccttct tcaggcaagt tcaatgacaa
     4861 tttcaacatc attgcagcag acaagatagt ggcgataggg ttgaccttat tctttggcaa
     4921 atctggagca gaaccgtggc atggttcgta caaaccaaat gcggtgttct tgtctggcaa
     4981 agaggccaag gacgcagatg gcaacaaacc caaggaacct gggataacgg aggcttcatc
     5041 ggagatgata tcaccaaaca tgttgctggt gattataata ccatttaggt gggttgggtt
     5101 cttaactagg atcatggcgg cagaatcaat caattgatgt tgaaccttca atgtaggaaa
     5161 ttcgttcttg atggtttcct ccacagtttt tctccataat cttgaagagg ccaaaacatt
     5221 agctttatcc aaggaccaaa taggcaatgg tggctcatgt tgtagggcca tgaaagcggc
     5281 cattcttgtg attctttgca cttctggaac ggtgtattgt tcactatccc aagcgacacc
     5341 atcaccatcg tcttcctttc tcttaccaaa gtaaatacct cccactaatt ctctgacaac
     5401 aacgaagtca gtacctttag caaattgtgg cttgattgga gataagtcta aaagagagtc
     5461 ggatgcaaag ttacatggtc ttaagttggc gtacaattga agttctttac ggatttttag
     5521 taaaccttgt tcaggtctaa cactacctgt accccattta ggaccaccca cagcacctaa
     5581 caaaacggca tcaaccttct tggaggcttc cagcgcctca tctggaagtg ggacacctgt
     5641 agcgtcgata gcagcaccac caattaaatg attttcgaaa tcgaacttga cattggaacg
     5701 aacatcagaa atagctttaa gaaccttaat ggcttcggct gtgatttctt gaccaacgtg
     5761 gtcacctggc aaaacgacga tcttcttagg ggcagacatt agaatggtat atccttgaaa
     5821 tatatatata tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct
     5881 aaccacctat tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg
     5941 atgcaagcat ttagtcatga acgcttctct attctatatg aaaagccggt tccggcgctc
     6001 tcacctttcc tttttctccc aatttttcag ttgaaaaagg tatatgcgtc aggcgacctc
     6061 tgaaattaac aaaaaatttc cagtcatcga atttgattct gtgcgatagc gcccctgtgt
     6121 gttctcgtta tgttgaggaa aaaaataatg gttgctaaga gattcgaact cttgcatctt
     6181 acgatacctg agtattccca cagttgggga tctcgactct agctagagga tcaattcgta
     6241 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6301 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6361 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgataac
     6421 ttcgtataat gtatgctata cgaagttatt aggtctgaag aggagtttac gtccagccaa
     6481 gctagcttgg ctgcaggtcg agcggccgcg atccggaacc cttaatataa cttcgtataa
     6541 tgtatgctat acgaagttat cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6601 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6661 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6721 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6781 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6841 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6901 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6961 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7021 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7081 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7141 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7201 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     7261 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7321 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7381 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7441 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7501 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7561 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7621 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7681 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7741 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7801 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7861 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7921 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7981 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8041 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8101 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8161 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8221 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8281 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8341 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8401 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8461 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8521 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     8581 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     8641 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     8701 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     8761 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataacgca tttaagcata
     8821 aacacgcact atgccgttct tctcatgtat atatatatac aggcaacacg cagatatagg
     8881 tgcgacgtga acagtgagct gtatgtgcgc agctcgcgtt gcattttcgg aagcgctcgt
     8941 tttcggaaac gctttgaagt tcctattccg aagttcctat tctctagcta gaaagtatag
     9001 gaacttcaga gcgcttttga aaaccaaaag cgctctgaag acgcactttc aaaaaaccaa
     9061 aaacgcaccg gactgtaacg agctactaaa atattgcgaa taccgcttcc acaaacattg
     9121 ctcaaaagta tctctttgct atatatctct gtgctatatc cctatataac ctacccatcc
     9181 acctttcgct ccttgaactt gcatctaaac tcgacctcta cattttttat gtttatctct
     9241 agtattactc tttagacaaa aaaattgtag taagaactat tcatagagtg aatcgaaaac
     9301 aatacgaaaa tgtaaacatt tcctatacgt agtatataga gacaaaatag aagaaaccgt
     9361 tcataatttt ctgaccaatg aagaatcatc aacgctatca ctttctgttc acaaagtatg
     9421 cgcaatccac atcggtatag aatataatcg gggatgcctt tatcttgaaa aaatgcaccc
     9481 gcagcttcgc tagtaatcag taaacgcggg aagtggagtc aggctttttt tatggaagag
     9541 aaaatagaca ccaaagtagc cttcttctaa ccttaacgga cctacagtgc aaaaagttat
     9601 caagagactg cattatagag cgcacaaagg agaaaaaaag taatctaaga tgctttgtta
     9661 gaaaaatagc gctctcggga tgcatttttg tagaacaaaa aagaagtata gattctttgt
     9721 tggtaaaata gcgctctcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt
     9781 gtttgaaaaa ttagcgctct cgcgttgcat ttttgtttta caaaaatgaa gcacagattc
     9841 ttcgttggta aaatagcgct ttcgcgttgc atttctgttc tgtaaaaatg cagctcagat
     9901 tctttgtttg aaaaattagc gctctcgcgt tgcatttttg ttctacaaaa tgaagcacag
     9961 atgcttcgtt gct
//
