LOCUS       62056_11220        9136 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 9136)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1478..1665
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(1785..2876)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIEATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(2877..3282)
                     /label=LEU2 promoter
     primer_bind     complement(3324..3340)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3348..3364)
                     /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(3372..3402)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3417..3438)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     protein_bind    3493..3526
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(3877..4465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4639..5496)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5497..5601)
                     /label=AmpR promoter
     rep_origin      5883..7047
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        7821..8525
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             8571..8591
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             8607..8948
                     /codon_start=1
                     /label=GAL4 activation domain
                     /note="activation domain of the GAL4 transcriptional
                     activator"
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     promoter        8954..8972
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             8991..9017
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             join(9045..9136,1..1087)
                     /codon_start=1
                     /label=p53
                     /note="human tumor suppressor protein that is often mutated
                     in cancers"
                     /translation="MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLML
                     SPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQ
                     GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY
                     KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
ORIGIN
        1 tctgtccccc ttgccgtccc aagcaatgga tgatttgatg ctgtccccgg acgatattga
       61 acaatggttc actgaagacc caggtccaga tgaagctccc agaatgccag aggctgctcc
      121 ccccgtggcc cctgcaccag cagctcctac accggcggcc cctgcaccag ccccctcctg
      181 gcccctgtca tcttctgtcc cttcccagaa aacctaccag ggcagctacg gtttccgtct
      241 gggcttcttg cattctggga cagccaagtc tgtgacttgc acgtactccc ctgccctcaa
      301 caagatgttt tgccaactgg ccaagacctg ccctgtgcag ctgtgggttg attccacacc
      361 cccgcccggc acccgcgtcc gcgccatggc catctacaag cagtcacagc acatgacgga
      421 ggttgtgagg cgctgccccc accatgagcg ctgctcagat agcgatggtc tggcccctcc
      481 tcagcatctt atccgagtgg aaggaaattt gcgtgtggag tatttggatg acagaaacac
      541 ttttcgacat agtgtggtgg tgccctatga gccgcctgag gttggctctg actgtaccac
      601 catccactac aactacatgt gtaacagttc ctgcatgggc ggcatgaacc ggaggcccat
      661 cctcaccatc atcacactgg aagactccag tggtaatcta ctgggacgga acagctttga
      721 ggtgcgtgtt tgtgcctgtc ctgggagaga ccggcgcaca gaggaagaga atctccgcaa
      781 gaaaggggag cctcaccacg agctgccccc agggagcact aagcgagcac tgcccaacaa
      841 caccagctcc tctccccagc caaagaagaa accactggat ggagaatatt tcacccttca
      901 gatccgtggg cgtgagcgct tcgagatgtt ccgagagctg aatgaggcct tggaactcaa
      961 ggatgcccag gctgggaagg agccaggggg gagcagggct cactccagcc acctgaagtc
     1021 caaaaagggt cagtctacct cccgccataa aaaactcatg ttcaagacag aagggcctga
     1081 ctcagactga actcgagctg cagatgaatc gtagatactg aaaaaccccg caagttcact
     1141 tcaactgtgc atcgtgcacc atctcaattt ctttcattta tacatcgttt tgccttcttt
     1201 tatgtaacta tactcctcta agtttcaatc ttggccatgt aacctctgat ctatagaatt
     1261 ttttaaatga ctagaattaa tgcccatctt ttttttggac ctaaattctt catgaaaata
     1321 tattacgagg gcttattcag aagctttgga cttcttcgcc agaggtttgg tcaagtctcc
     1381 aatcaaggtt gtcggcttgt ctaccttgcc agaaatttac gaaaagatgg aaaagggtca
     1441 aatcgttggt agatacgttg ttgacacttc taaataagcg aatttcttat gatttatgat
     1501 ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta
     1561 ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc
     1621 tcaggtatag catgaggtcg ctcttattga ccacacctct accggccggt cgaaattccc
     1681 ctaccctatg aacatattcc attttgtaat ttcgtgtcgt ttctattatg aatttcattt
     1741 ataaagttta tgtacaaata tcataaaaaa agagaatctt tttaagcaag gattttctta
     1801 acttcttcgg cgacagcatc accgacttcg gtggtactgt tggaaccacc taaatcacca
     1861 gttctgatac ctgcatccaa aaccttttta actgcatctt caatggcctt accttcttca
     1921 ggcaagttca atgacaattt caacatcatt gcagcagaca agatagtggc gatagggttg
     1981 accttattct ttggcaaatc tggagcagaa ccgtggcatg gttcgtacaa accaaatgcg
     2041 gtgttcttgt ctggcaaaga ggccaaggac gcagatggca acaaacccaa ggaacctggg
     2101 ataacggagg cttcatcgga gatgatatca ccaaacatgt tgctggtgat tataatacca
     2161 tttaggtggg ttgggttctt aactaggatc atggcggcag aatcaatcaa ttgatgttga
     2221 accttcaatg taggaaattc gttcttgatg gtttcctcca cagtttttct ccataatctt
     2281 gaagaggcca aaacattagc tttatccaag gaccaaatag gcaatggtgg ctcatgttgt
     2341 agggccatga aagcggccat tcttgtgatt ctttgcactt ctggaacggt gtattgttca
     2401 ctatcccaag cgacaccatc accatcgtct tcctttctct taccaaagta aatacctccc
     2461 actaattctc tgacaacaac gaagtcagta cctttagcaa attgtggctt gattggagat
     2521 aagtctaaaa gagagtcgga tgcaaagtta catggtctta agttggcgta caattgaagt
     2581 tctttacgga tttttagtaa accttgttca ggtctaacac tacctgtacc ccatttagga
     2641 ccacccacag cacctaacaa aacggcatca gccttcttgg aggcttccag cgcctcatct
     2701 ggaagtggga cacctgtagc ttcgatagca gcaccaccaa ttaaatgatt ttcgaaatcg
     2761 aacttgacat tggaacgaac atcagaaata gctttaagaa ccttaatggc ttcggctgtg
     2821 atttcttgac caacgtggtc acctggcaaa acgacgatct tcttaggggc agacattaga
     2881 atggtatatc cttgaaatat atatatatat tgctgaaatg taaaaggtaa gaaaagttag
     2941 aaagtaagac gattgctaac cacctattgg aaaaaacaat aggtccttaa ataatattgt
     3001 caacttcaag tattgtgatg caagcattta gtcatgaacg cttctctatt ctatatgaaa
     3061 agccggttcc ggcgctctca cctttccttt ttctcccaat ttttcagttg aaaaaggtat
     3121 atgcgtcagg cgacctctga aattaacaaa aaatttccag tcatcgaatt tgattctgtg
     3181 cgatagcgcc cctgtgtgtt ctcgttatgt tgaggaaaaa aataatggtt gctaagagat
     3241 tcgaactctt gcatcttacg atacctgagt attcccacag ttgggggatc tcgactctag
     3301 ctagaggatc aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc
     3361 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     3421 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3481 tcgtgccagc tgataacttc gtataatgta tgctatacga agttattagg tctgaagagg
     3541 agtttacgtc cagccaagct agcttggctg caggtcgagc ggccgcgatc cggaaccctt
     3601 aatataactt cgtataatgt atgctatacg aagttatcag ctgcattaat gaatcggcca
     3661 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3721 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3781 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3841 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3901 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3961 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4021 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4081 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4141 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4201 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4261 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4321 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4381 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4441 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4501 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4561 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4621 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4681 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4741 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4801 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4861 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4921 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4981 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5041 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5101 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5161 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5221 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5281 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5341 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5401 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5461 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5521 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5581 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5641 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     5701 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     5761 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     5821 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     5881 taacgcattt aagcataaac acgcactatg ccgttcttct catgtatata tatatacagg
     5941 caacacgcag atataggtgc gacgtgaaca gtgagctgta tgtgcgcagc tcgcgttgca
     6001 ttttcggaag cgctcgtttt cggaaacgct ttgaagttcc tattccgaag ttcctattct
     6061 ctagctagaa agtataggaa cttcagagcg cttttgaaaa ccaaaagcgc tctgaagacg
     6121 cactttcaaa aaaccaaaaa cgcaccggac tgtaacgagc tactaaaata ttgcgaatac
     6181 cgcttccaca aacattgctc aaaagtatct ctttgctata tatctctgtg ctatatccct
     6241 atataaccta cccatccacc tttcgctcct tgaacttgca tctaaactcg acctctacat
     6301 tttttatgtt tatctctagt attactcttt agacaaaaaa attgtagtaa gaactattca
     6361 tagagtgaat cgaaaacaat acgaaaatgt aaacatttcc tatacgtagt atatagagac
     6421 aaaatagaag aaaccgttca taattttctg accaatgaag aatcatcaac gctatcactt
     6481 tctgttcaca aagtatgcgc aatccacatc ggtatagaat ataatcgggg atgcctttat
     6541 cttgaaaaaa tgcacccgca gcttcgctag taatcagtaa acgcgggaag tggagtcagg
     6601 ctttttttat ggaagagaaa atagacacca aagtagcctt cttctaacct taacggacct
     6661 acagtgcaaa aagttatcaa gagactgcat tatagagcgc acaaaggaga aaaaaagtaa
     6721 tctaagatgc tttgttagaa aaatagcgct ctcgggatgc atttttgtag aacaaaaaag
     6781 aagtatagat tctttgttgg taaaatagcg ctctcgcgtt gcatttctgt tctgtaaaaa
     6841 tgcagctcag attctttgtt tgaaaaatta gcgctctcgc gttgcatttt tgttttacaa
     6901 aaatgaagca cagattcttc gttggtaaaa tagcgctttc gcgttgcatt tctgttctgt
     6961 aaaaatgcag ctcagattct ttgtttgaaa aattagcgct ctcgcgttgc atttttgttc
     7021 tacaaaatga agcacagatg cttcgttgct tgcatgcctg caggtcgaga tccgggatcg
     7081 aagaaatgat ggtaaatgaa ataggaaatc aaggagcatg aaggcaaaag acaaatataa
     7141 gggtcgaacg aaaaataaag tgaaaagtgt tgatatgatg tatttggctt tgcggcgccg
     7201 aaaaaacgag tttacgcaat tgcacaatca tgctgactct gtggcggacc cgcgctcttg
     7261 ccggcccggc gataacgctg ggcgtgaggc tgtgcccggc ggagtttttt gcgcctgcat
     7321 tttccaaggt ttaccctgcg ctaaggggcg agattggaga agcaataaga atgccggttg
     7381 gggttgcgat gatgacgacc acgacaactg gtgtcattat ttaagttgcc gaaagaacct
     7441 gagtgcattt gcaacatgag tatactagaa gaatgagcca agacttgcga gacgcgagtt
     7501 tgccggtggt gcgaacaata gagcgaccat gaccttgaag gtgagacgcg cataaccgct
     7561 agagtacttt gaagaggaaa cagcaatagg gttgctacca gtataaatag acaggtacat
     7621 acaacactgg aaatggttgt ctgtttgagt acgctttcaa ttcatttggg tgtgcacttt
     7681 attatgttac aatatggaag ggaactttac acttctccta tgcacatata ttaattaaag
     7741 tccaatgcta gtagagaagg ggggtaacac ccctccgcgc tcttttccga tttttttcta
     7801 aaccgtggaa tatttcggat atccttttgt tgtttccggg tgtacaatat ggacttcctc
     7861 ttttctggca accaaaccca tacatcggga ttcctataat accttcgttg gtctccctaa
     7921 catgtaggtg gcggagggga gatatacaat agaacagata ccagacaaga cataatgggc
     7981 taaacaagac tacaccaatt acactgcctc attgatggtg gtacataacg aactaatact
     8041 gtagccctag acttgatagc catcatcata tcgaagtttc actacccttt ttccatttgc
     8101 catctattga agtaataata ggcgcatgca acttcttttc tttttttttc ttttctctct
     8161 cccccgttgt tgtctcacca tatccgcaat gacaaaaaaa tgatggaaga cactaaagga
     8221 aaaaattaac gacaaagaca gcaccaacag atgtcgttgt tccagagctg atgaggggta
     8281 tctcgaagca cacgaaactt tttccttcct tcattcacgc acactactct ctaatgagca
     8341 acggtatacg gccttccttc cagttacttg aatttgaaat aaaaaaaagt ttgctgtctt
     8401 gctatcaagt ataaatagac ctgcaattat taatcttttg tttcctcgtc attgttctcg
     8461 ttccctttct tccttgtttc tttttctgca caatatttca agctatacca agcatacaat
     8521 caactccaag ctttgcaaag atggataaag cggaattaat tcccgagcct ccaaaaaaga
     8581 agagaaaggt cgaattgggt accgccgcca attttaatca aagtgggaat attgctgata
     8641 gctcattgtc cttcactttc actaacagta gcaacggtcc gaacctcata acaactcaaa
     8701 caaattctca agcgctttca caaccaattg cctcctctaa cgttcatgat aacttcatga
     8761 ataatgaaat cacggctagt aaaattgatg atggtaataa ttcaaaacca ctgtcacctg
     8821 gttggacgga ccaaactgcg tataacgcgt ttggaatcac tacagggatg tttaatacca
     8881 ctacaatgga tgatgtatat aactatctat tcgatgatga agatacccca ccaaacccaa
     8941 aaaaagagat ctttaatacg actcactata gggcgagcgc cgccatggag tacccatacg
     9001 acgtaccaga ttacgctcat atggccatgg aggccagtga attcatggag gagccgcagt
     9061 cagatcctag cgtcgagccc cctctgagtc aggaaacatt ttcagaccta tggaaactac
     9121 ttcctgaaaa caacgt
//
