LOCUS       40924_20930        6659 bp DNA     circular SYN 13-JAN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6659)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6659)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..406
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             452..472
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             488..829
                     /codon_start=1
                     /label=GAL4 activation domain
                     /note="activation domain of the GAL4 transcriptional
                     activator"
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     terminator      1241..1428
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(1549..2640)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(2641..3045)
                     /label=LEU2 promoter
     primer_bind     complement(3087..3103)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3111..3127)
                     /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(3135..3165)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3180..3201)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3489..4077)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4251..5108)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5109..5213)
                     /label=AmpR promoter
     rep_origin      5495..6659
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gcttgcatgc aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc
       61 atatccgcaa tgacaaaaaa aatgatggaa gacactaaag gaaaaaatta acgacaaaga
      121 cagcaccaac agatgtcgtt gttccagagc tgatgagggg tatcttcgaa cacacgaaac
      181 tttttccttc cttcattcac gcacactact ctctaatgag caacggtata cggccttcct
      241 tccagttact tgaatttgaa ataaaaaaag tttgccgctt tgctatcaag tataaataga
      301 cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt
      361 ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactccaa gctttgcaaa
      421 gatggataaa gcggaattaa ttcccgagcc tccaaaaaag aagagaaagg tcgaattggg
      481 taccgccgcc aattttaatc aaagtgggaa tattgctgat agctcattgt ccttcacttt
      541 cactaacagt agcaacggtc cgaacctcat aacaactcaa acaaattctc aagcgctttc
      601 acaaccaatt gcctcctcta acgttcatga taacttcatg aataatgaaa tcacggctag
      661 taaaattgat gatggtaata attcaaaacc actgtcacct ggttggacgg accaaactgc
      721 gtataacgcg tttggaatca ctacagggat gtttaatacc actacaatgg atgatgtata
      781 taactatcta ttcgatgatg aagatacccc accaaaccca aaaaaagaga tcgaattccc
      841 ggggatccgt cgacctgcag agatctatga atcgtagata ctgaaaaacc ccgcaagttc
      901 acttcaactg tgcatcgtgc accatctcaa tttctttcat ttatacatcg ttttgccttc
      961 ttttatgtaa ctatactcct ctaagtttca atcttggcca tgtaacctct gatctataga
     1021 attttttaaa tgactagaat taatgcccat cttttttttg gacctaaatt cttcatgaaa
     1081 atatattacg agggcttatt cagaagcttt ggacttcttc gccagaggtt tggtcaagtc
     1141 tccaatcaag gttgtcggct tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg
     1201 tcaaatcgtt ggtagatacg ttgttgacac ttctaaataa gcgaatttct tatgatttat
     1261 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     1321 ttaggtttta aaacgaaaat tcttgttctt gagtaactct ttcctgtagg tcaggttgct
     1381 ttctcaggta tagcatgagg tcgctcttat tgaccacacc tctaccggca tgcccgaaat
     1441 tcccctaccc tatgaacata ttccattttg taatttcgtg tcgtttctat tatgaatttc
     1501 atttataaag tttatgtaca aatatcataa aaaaagagaa tctttttaag caaggatttt
     1561 cttaacttct tcggcgacag catcaccgac ttcggtggta ctgttggaac cacctaaatc
     1621 accagttctg atacctgcat ccaaaacctt tttaactgca tcttcaatgg ccttaccttc
     1681 ttcaggcaag ttcaatgaca atttcaacat cattgcagca gacaagatag tggcgatagg
     1741 gtcaacctta ttctttggca aatctggagc agaaccgtgg catggttcgt acaaaccaaa
     1801 tgcggtgttc ttgtctggca aagaggccaa ggacgcagat ggcaacaaac ccaaggaacc
     1861 tgggataacg gaggcttcat cggagatgat atcaccaaac atgttgctgg tgattataat
     1921 accatttagg tgggttgggt tcttaactag gatcatggcg gcagaatcaa tcaattgatg
     1981 ttgaaccttc aatgtaggaa attcgttctt gatggtttcc tccacagttt ttctccataa
     2041 tcttgaagag gccaaaacat tagctttatc caaggaccaa ataggcaatg gtggctcatg
     2101 ttgtagggcc atgaaagcgg ccattcttgt gattctttgc acttctggaa cggtgtattg
     2161 ttcactatcc caagcgacac catcaccatc gtcttccttt ctcttaccaa agtaaatacc
     2221 tcccactaat tctctgacaa caacgaagtc agtaccttta gcaaattgtg gcttgattgg
     2281 agataagtct aaaagagagt cggatgcaaa gttacatggt cttaagttgg cgtacaattg
     2341 aagttcttta cggattttta gtaaaccttg ttcaggtcta acactacctg taccccattt
     2401 aggaccaccc acagcaccta acaaaacggc atcaaccttc ttggaggctt ccagcgcctc
     2461 atctggaagt gggacacctg tagcatcgat agcagcacca ccaattaaat gattttcgaa
     2521 atcgaacttg acattggaac gaacatcaga aatagcttta agaaccttaa tggcttcggc
     2581 tgtgatttct tgaccaacgt ggtcacctgg caaaacgacg atcttcttag gggcagacat
     2641 tagaatggta tatccttgaa atatatatat atattgctga aatgtaaaag gtaagaaaag
     2701 ttagaaagta agacgattgc taaccaccta ttggaaaaaa caataggtcc ttaaataata
     2761 ttgtcaactt caagtattgt gatgcaagca tttagtcatg aacgcttctc tattctatat
     2821 gaaaagccgg ttccggcctc tcacctttcc tttttctccc aatttttcag ttgaaaaagg
     2881 tatatgcgtc aggcgacctc tgaaattaac aaaaaatttc cagtcatcga atttgattct
     2941 gtgcgatagc gcccctgtgt gttctcgtta tgttgaggaa aaaaataatg gttgctaaga
     3001 gattcgaact cttgcatctt acgatacctg agtattccca cagttgggga tctcgactct
     3061 agctagagga tcaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3121 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3181 tgagtgaggt aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3241 ctgtcgtgcc agctggatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3301 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3361 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3421 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3481 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3541 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3601 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3661 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3721 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3781 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3841 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3901 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     3961 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4021 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4081 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4141 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4201 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4261 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4321 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4381 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4441 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4501 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4561 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4621 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4681 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4741 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4801 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4861 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4921 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4981 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5041 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5101 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5161 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5221 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     5281 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     5341 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     5401 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     5461 gcatcagagc agattgtact gagagtgcac cataacgcat ttaagcataa acacgcacta
     5521 tgccgttctt ctcatgtata tatatataca ggcaacacgc agatataggt gcgacgtgaa
     5581 cagtgagctg tatgtgcgca gctcgcgttg cattttcgga agcgctcgtt ttcggaaacg
     5641 ctttgaagtt cctattccga agttcctatt ctctagctag aaagtatagg aacttcagag
     5701 cgcttttgaa aaccaaaagc gctctgaaga cgcactttca aaaaaccaaa aacgcaccgg
     5761 actgtaacga gctactaaaa tattgcgaat accgcttcca caaacattgc tcaaaagtat
     5821 ctctttgcta tatatctctg tgctatatcc ctatataacc tacccatcca cctttcgctc
     5881 cttgaacttg catctaaact cgacctctac attttttatg tttatctcta gtattactct
     5941 ttagacaaaa aaattgtagt aagaactatt catagagtga atcgaaaaca atacgaaaat
     6001 gtaaacattt cctatacgta gtatatagag acaaaataga agaaaccgtt cataattttc
     6061 tgaccaatga agaatcatca acgctatcac tttctgttca caaagtatgc gcaatccaca
     6121 tcggtataga atataatcgg ggatgccttt atcttgaaaa aatgcacccg cagcttcgct
     6181 agtaatcagt aaacgcggga agtggagtca ggcttttttt atggaagaga aaatagacac
     6241 caaagtagcc ttcttctaac cttaacggac ctacagtgca aaaagttatc aagagactgc
     6301 attatagagc gcacaaagga gaaaaaaagt aatctaagat gctttgttag aaaaatagcg
     6361 ctctcgggat gcatttttgt agaacaaaaa agaagtatag attctttgtt ggtaaaatag
     6421 cgctctcgcg ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat
     6481 tagcgctctc gcgttgcatt tttgttttac aaaaatgaag cacagattct tcgttggtaa
     6541 aatagcgctt tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga
     6601 aaaattagcg ctctcgcgtt gcatttttgt tctacaaaat gaagcacaga tgcttcgtt
//
