LOCUS       62056_11210        8407 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 8407)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8407
                     /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      2835..3022
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(3142..4233)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4234..4639)
                     /label=LEU2 promoter
     primer_bind     complement(4681..4697)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4705..4721)
                     /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(4729..4759)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4774..4795)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     protein_bind    4850..4883
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(5234..5822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5996..6853)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6854..6958)
                     /label=AmpR promoter
     rep_origin      7240..8404
                     /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 tatgatgttc
     1981 agtacacaat cgcaagccat ggattcacga attgcagtag tctgtgttgt tttggcagtg
     2041 ttcgcggtgg accagacggt cggagcgccg caaaaagatg ctgtggcagc aagcggtccc
     2101 gcctacacca caaaatacga ccatattgac atcgaccaag ttttgggttc caaaagatta
     2161 gtaaacagct acgttcaatg tctgctggac aagaaaccgt gcactcccga aggagctgaa
     2221 cttagaaaaa ttttacctga tgccttaaaa acccaatgtg tgaaatgcaa tgctacgcaa
     2281 aaaaatgctg ctcttaaagt agttgaccga ctccagagag attatgataa agaatggaaa
     2341 cagcttcttg acaaatggga ccctaagcgt gaatatttcc agaaattcca acaatttttg
     2401 gcagaagaga agaaaaaagg tgttgtcaaa ttttaaggat ccatcgagct cgagctgcag
     2461 atgaatcgta gatactgaaa aaccccgcaa gttcacttca actgtgcatc gtgcaccatc
     2521 tcaatttctt tcatttatac atcgttttgc cttcttttat gtaactatac tcctctaagt
     2581 ttcaatcttg gccatgtaac ctctgatcta tagaattttt taaatgacta gaattaatgc
     2641 ccatcttttt tttggaccta aattcttcat gaaaatatat tacgagggct tattcagaag
     2701 ctttggactt cttcgccaga ggtttggtca agtctccaat caaggttgtc ggcttgtcta
     2761 ccttgccaga aatttacgaa aagatggaaa agggtcaaat cgttggtaga tacgttgttg
     2821 acacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
     2881 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
     2941 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat gaggtcgctc
     3001 ttattgacca cacctctacc ggccggtcga aattccccta ccctatgaac atattccatt
     3061 ttgtaatttc gtgtcgtttc tattatgaat ttcatttata aagtttatgt acaaatatca
     3121 taaaaaaaga gaatcttttt aagcaaggat tttcttaact tcttcggcga cagcatcacc
     3181 gacttcggtg gtactgttgg aaccacctaa atcaccagtt ctgatacctg catccaaaac
     3241 ctttttaact gcatcttcaa tggccttacc ttcttcaggc aagttcaatg acaatttcaa
     3301 catcattgca gcagacaaga tagtggcgat agggttgacc ttattctttg gcaaatctgg
     3361 agcagaaccg tggcatggtt cgtacaaacc aaatgcggtg ttcttgtctg gcaaagaggc
     3421 caaggacgca gatggcaaca aacccaagga acctgggata acggaggctt catcggagat
     3481 gatatcacca aacatgttgc tggtgattat aataccattt aggtgggttg ggttcttaac
     3541 taggatcatg gcggcagaat caatcaattg atgttgaacc ttcaatgtag gaaattcgtt
     3601 cttgatggtt tcctccacag tttttctcca taatcttgaa gaggccaaaa cattagcttt
     3661 atccaaggac caaataggca atggtggctc atgttgtagg gccatgaaag cggccattct
     3721 tgtgattctt tgcacttctg gaacggtgta ttgttcacta tcccaagcga caccatcacc
     3781 atcgtcttcc tttctcttac caaagtaaat acctcccact aattctctga caacaacgaa
     3841 gtcagtacct ttagcaaatt gtggcttgat tggagataag tctaaaagag agtcggatgc
     3901 aaagttacat ggtcttaagt tggcgtacaa ttgaagttct ttacggattt ttagtaaacc
     3961 ttgttcaggt ctaacactac ctgtacccca tttaggacca cccacagcac ctaacaaaac
     4021 ggcatcaacc ttcttggagg cttccagcgc ctcatctgga agtgggacac ctgtagcgtc
     4081 gatagcagca ccaccaatta aatgattttc gaaatcgaac ttgacattgg aacgaacatc
     4141 agaaatagct ttaagaacct taatggcttc ggctgtgatt tcttgaccaa cgtggtcacc
     4201 tggcaaaacg acgatcttct taggggcaga cattagaatg gtatatcctt gaaatatata
     4261 tatatattgc tgaaatgtaa aaggtaagaa aagttagaaa gtaagacgat tgctaaccac
     4321 ctattggaaa aaacaatagg tccttaaata atattgtcaa cttcaagtat tgtgatgcaa
     4381 gcatttagtc atgaacgctt ctctattcta tatgaaaagc cggttccggc gctctcacct
     4441 ttcctttttc tcccaatttt tcagttgaaa aaggtatatg cgtcaggcga cctctgaaat
     4501 taacaaaaaa tttccagtca tcgaatttga ttctgtgcga tagcgcccct gtgtgttctc
     4561 gttatgttga ggaaaaaaat aatggttgct aagagattcg aactcttgca tcttacgata
     4621 cctgagtatt cccacagttg gggatctcga ctctagctag aggatcaatt cgtaatcatg
     4681 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4741 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4801 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctga taacttcgta
     4861 taatgtatgc tatacgaagt tattaggtct gaagaggagt ttacgtccag ccaagctagc
     4921 ttggctgcag gtcgagcggc cgcgatccgg aacccttaat ataacttcgt ataatgtatg
     4981 ctatacgaag ttatcagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5041 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5101 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5161 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5221 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5281 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5341 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5401 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5461 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5521 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5581 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5641 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     5701 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5761 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5821 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5881 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5941 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6001 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6061 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6121 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6181 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6241 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6301 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6361 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6421 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6481 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6541 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6601 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6661 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6721 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6781 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6841 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6901 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6961 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     7021 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     7081 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     7141 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     7201 atgcggcatc agagcagatt gtactgagag tgcaccataa cgcatttaag cataaacacg
     7261 cactatgccg ttcttctcat gtatatatat atacaggcaa cacgcagata taggtgcgac
     7321 gtgaacagtg agctgtatgt gcgcagctcg cgttgcattt tcggaagcgc tcgttttcgg
     7381 aaacgctttg aagttcctat tccgaagttc ctattctcta gctagaaagt ataggaactt
     7441 cagagcgctt ttgaaaacca aaagcgctct gaagacgcac tttcaaaaaa ccaaaaacgc
     7501 accggactgt aacgagctac taaaatattg cgaataccgc ttccacaaac attgctcaaa
     7561 agtatctctt tgctatatat ctctgtgcta tatccctata taacctaccc atccaccttt
     7621 cgctccttga acttgcatct aaactcgacc tctacatttt ttatgtttat ctctagtatt
     7681 actctttaga caaaaaaatt gtagtaagaa ctattcatag agtgaatcga aaacaatacg
     7741 aaaatgtaaa catttcctat acgtagtata tagagacaaa atagaagaaa ccgttcataa
     7801 ttttctgacc aatgaagaat catcaacgct atcactttct gttcacaaag tatgcgcaat
     7861 ccacatcggt atagaatata atcggggatg cctttatctt gaaaaaatgc acccgcagct
     7921 tcgctagtaa tcagtaaacg cgggaagtgg agtcaggctt tttttatgga agagaaaata
     7981 gacaccaaag tagccttctt ctaaccttaa cggacctaca gtgcaaaaag ttatcaagag
     8041 actgcattat agagcgcaca aaggagaaaa aaagtaatct aagatgcttt gttagaaaaa
     8101 tagcgctctc gggatgcatt tttgtagaac aaaaaagaag tatagattct ttgttggtaa
     8161 aatagcgctc tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga
     8221 aaaattagcg ctctcgcgtt gcatttttgt tttacaaaaa tgaagcacag attcttcgtt
     8281 ggtaaaatag cgctttcgcg ttgcatttct gttctgtaaa aatgcagctc agattctttg
     8341 tttgaaaaat tagcgctctc gcgttgcatt tttgttctac aaaatgaagc acagatgctt
     8401 cgttgct
//
