LOCUS       62056_11215        8413 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 8413)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8413
                     /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"
     terminator      2841..3028
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(3148..4239)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4240..4645)
                     /label=LEU2 promoter
     primer_bind     complement(4687..4703)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4711..4727)
                     /label=lac operator
                     /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(4735..4765)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4780..4801)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     protein_bind    4856..4889
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(5240..5828)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6002..6859)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6860..6964)
                     /label=AmpR promoter
     rep_origin      7246..8410
                     /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 tatgatggct
     1981 ttcaaggaaa tcgcaatgtt ttccagttta gtggtggtca tcataagcta caatgtacta
     2041 gaaagcaatg cattaatgat gcaaccatta atagaccaag atttctgtgc agtaaaagct
     2101 agtttgtacg aattgggaga ggctgagatg aacttgatgg acgattcgca gacattaaac
     2161 gacatgcaaa gagaatattt tgctggaaaa gttgactatt ctgctgtgga aagagcaagg
     2221 aatgaactta ataagtccaa aaacaagtgg ttccaaaaag ttatacaata tctcaaggca
     2281 acaaatgaat ttgagcccac aatcaactat caaacctcag atccacaaaa agtttaccat
     2341 caaatggatg acttggacaa ttataaagat gctctagatg atatgcatga aaatatcctt
     2401 aaatcaatgt cccctaccgt agcccaacct gttgtagcat aaggatccat cgagctcgag
     2461 ctgcagatga atcgtagata ctgaaaaacc ccgcaagttc acttcaactg tgcatcgtgc
     2521 accatctcaa tttctttcat ttatacatcg ttttgccttc ttttatgtaa ctatactcct
     2581 ctaagtttca atcttggcca tgtaacctct gatctataga attttttaaa tgactagaat
     2641 taatgcccat cttttttttg gacctaaatt cttcatgaaa atatattacg agggcttatt
     2701 cagaagcttt ggacttcttc gccagaggtt tggtcaagtc tccaatcaag gttgtcggct
     2761 tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg tcaaatcgtt ggtagatacg
     2821 ttgttgacac ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag
     2881 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
     2941 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg
     3001 tcgctcttat tgaccacacc tctaccggcc ggtcgaaatt cccctaccct atgaacatat
     3061 tccattttgt aatttcgtgt cgtttctatt atgaatttca tttataaagt ttatgtacaa
     3121 atatcataaa aaaagagaat ctttttaagc aaggattttc ttaacttctt cggcgacagc
     3181 atcaccgact tcggtggtac tgttggaacc acctaaatca ccagttctga tacctgcatc
     3241 caaaaccttt ttaactgcat cttcaatggc cttaccttct tcaggcaagt tcaatgacaa
     3301 tttcaacatc attgcagcag acaagatagt ggcgataggg ttgaccttat tctttggcaa
     3361 atctggagca gaaccgtggc atggttcgta caaaccaaat gcggtgttct tgtctggcaa
     3421 agaggccaag gacgcagatg gcaacaaacc caaggaacct gggataacgg aggcttcatc
     3481 ggagatgata tcaccaaaca tgttgctggt gattataata ccatttaggt gggttgggtt
     3541 cttaactagg atcatggcgg cagaatcaat caattgatgt tgaaccttca atgtaggaaa
     3601 ttcgttcttg atggtttcct ccacagtttt tctccataat cttgaagagg ccaaaacatt
     3661 agctttatcc aaggaccaaa taggcaatgg tggctcatgt tgtagggcca tgaaagcggc
     3721 cattcttgtg attctttgca cttctggaac ggtgtattgt tcactatccc aagcgacacc
     3781 atcaccatcg tcttcctttc tcttaccaaa gtaaatacct cccactaatt ctctgacaac
     3841 aacgaagtca gtacctttag caaattgtgg cttgattgga gataagtcta aaagagagtc
     3901 ggatgcaaag ttacatggtc ttaagttggc gtacaattga agttctttac ggatttttag
     3961 taaaccttgt tcaggtctaa cactacctgt accccattta ggaccaccca cagcacctaa
     4021 caaaacggca tcaaccttct tggaggcttc cagcgcctca tctggaagtg ggacacctgt
     4081 agcgtcgata gcagcaccac caattaaatg attttcgaaa tcgaacttga cattggaacg
     4141 aacatcagaa atagctttaa gaaccttaat ggcttcggct gtgatttctt gaccaacgtg
     4201 gtcacctggc aaaacgacga tcttcttagg ggcagacatt agaatggtat atccttgaaa
     4261 tatatatata tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct
     4321 aaccacctat tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg
     4381 atgcaagcat ttagtcatga acgcttctct attctatatg aaaagccggt tccggcgctc
     4441 tcacctttcc tttttctccc aatttttcag ttgaaaaagg tatatgcgtc aggcgacctc
     4501 tgaaattaac aaaaaatttc cagtcatcga atttgattct gtgcgatagc gcccctgtgt
     4561 gttctcgtta tgttgaggaa aaaaataatg gttgctaaga gattcgaact cttgcatctt
     4621 acgatacctg agtattccca cagttgggga tctcgactct agctagagga tcaattcgta
     4681 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4741 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4801 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgataac
     4861 ttcgtataat gtatgctata cgaagttatt aggtctgaag aggagtttac gtccagccaa
     4921 gctagcttgg ctgcaggtcg agcggccgcg atccggaacc cttaatataa cttcgtataa
     4981 tgtatgctat acgaagttat cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5041 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5101 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5161 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5221 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5281 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5341 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5401 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5461 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5521 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5581 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5641 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     5701 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5761 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5821 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5881 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5941 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6001 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6061 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6121 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6301 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6481 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6841 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6961 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     7021 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     7081 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     7141 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     7201 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataacgca tttaagcata
     7261 aacacgcact atgccgttct tctcatgtat atatatatac aggcaacacg cagatatagg
     7321 tgcgacgtga acagtgagct gtatgtgcgc agctcgcgtt gcattttcgg aagcgctcgt
     7381 tttcggaaac gctttgaagt tcctattccg aagttcctat tctctagcta gaaagtatag
     7441 gaacttcaga gcgcttttga aaaccaaaag cgctctgaag acgcactttc aaaaaaccaa
     7501 aaacgcaccg gactgtaacg agctactaaa atattgcgaa taccgcttcc acaaacattg
     7561 ctcaaaagta tctctttgct atatatctct gtgctatatc cctatataac ctacccatcc
     7621 acctttcgct ccttgaactt gcatctaaac tcgacctcta cattttttat gtttatctct
     7681 agtattactc tttagacaaa aaaattgtag taagaactat tcatagagtg aatcgaaaac
     7741 aatacgaaaa tgtaaacatt tcctatacgt agtatataga gacaaaatag aagaaaccgt
     7801 tcataatttt ctgaccaatg aagaatcatc aacgctatca ctttctgttc acaaagtatg
     7861 cgcaatccac atcggtatag aatataatcg gggatgcctt tatcttgaaa aaatgcaccc
     7921 gcagcttcgc tagtaatcag taaacgcggg aagtggagtc aggctttttt tatggaagag
     7981 aaaatagaca ccaaagtagc cttcttctaa ccttaacgga cctacagtgc aaaaagttat
     8041 caagagactg cattatagag cgcacaaagg agaaaaaaag taatctaaga tgctttgtta
     8101 gaaaaatagc gctctcggga tgcatttttg tagaacaaaa aagaagtata gattctttgt
     8161 tggtaaaata gcgctctcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt
     8221 gtttgaaaaa ttagcgctct cgcgttgcat ttttgtttta caaaaatgaa gcacagattc
     8281 ttcgttggta aaatagcgct ttcgcgttgc atttctgttc tgtaaaaatg cagctcagat
     8341 tctttgtttg aaaaattagc gctctcgcgt tgcatttttg ttctacaaaa tgaagcacag
     8401 atgcttcgtt gct
//
