LOCUS       Exported                7654 bp ds-DNA     circular SYN 03-MAR-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7654)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 4, 2019 from SnapGene Viewer 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7654
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    11..514
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        complement(569..587)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(608..624)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    632..648
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(656..686)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(1010..1598)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1769..2629)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2630..2734)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2737..2787
                     /label=oRH5050
     CDS             complement(2785..4119)
                     /codon_start=1
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /product="O-acetylserine and O-acetylhomoserine 
                     sulfhydrylase, required for sulfur amino acid synthesis"
                     /label=MET17
                     /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
     promoter        complement(4120..4353)
                     /label=MET17 Promoter
     primer_bind     complement(4334..4381)
                     /label=oRH5049
     primer_bind     4384..4403
                     /label=oRH4872
     promoter        4602..4977
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             4978..6693
                     /codon_start=1
                     /gene="S. cerevisiae ADE2"
                     /product="phosphoribosylaminoimidazole carboxylase, 
                     required for adenine biosynthesis"
                     /label=ADE2
                     /note="yeast auxotrophic marker"
                     /translation="MDSRTVGILGGGQLGRMIVEAANRLNIKTVILDAENSPAKQISNS
                     NDHVNGSFSNPLDIEKLAEKCDVLTIEIEHVDVPTLKNLQVKHPKLKIYPSPETIRLIQ
                     DKYIQKEHLIKNGIAVTQSVPVEQASETSLLNVGRDLGFPFVLKSRTLAYDGRGNFVVK
                     NKEMIPEALEVLKDRPLYAEKWAPFTKELAVMIVRSVNGLVFSYPIVETIHKDNICDLC
                     YAPARVPDSVQLKAKLLAENAIKSFPGCGIFGVEMFYLETGELLINEIAPRPHNSGHYT
                     IDACVTSQFEAHLRSILDLPMPKNFTSFSTITTNAIMLNVLGDKHTKDKELETCERALA
                     TPGSSVYLYGKESRPNRKVGHINIIASSMAECEQRLNYITGRTDIPIKISVAQKLDLEA
                     MVKPLVGIIMGSDSDLPVMSAACAVLKDFGVPFEVTIVSAHRTPHRMSAYAISASKRGI
                     KTIIAGAGGAAHLPGMVAAMTPLPVIGVPVKGSCLDGVDSLHSIVQMPRGVPVATVAIN
                     NSTNAALLAVRLLGAYDSSYTTKMEQFLLKQEEEVLVKAQKLETVGYEAYLENK"
     rep_origin      complement(6898..7353)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7012..7031)
                     /label=oRH4736
     primer_bind     7498..7514
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7517..7543
                     /label=Recomb OH 1
     promoter        7524..7542
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     7568..7584
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7618..7634)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(join(7634..7654,1..9))
                     /label=Recomb OH 2
ORIGIN
        1 tcttaggacg gatcgcttgc ctgtaactta cacgcgcctc gtatctttta atgatggaat
       61 aatttgggaa tttactctgt gtttatttat ttttatgttt tgtatttgga ttttagaaag
      121 taaataaaga aggtagaaga gttacggaat gaagaaaaaa aaataaacaa aggtttaaaa
      181 aatttcaaca aaaagcgtac tttacatata tatttattag acaagaaaag cagattaaat
      241 agatatacat tcgattaacg ataagtaaaa tgtaaaatca caggattttc gtgtgtggtc
      301 ttctacacag acaagatgaa acaattcggc attaatacct gagagcagga agagcaagat
      361 aaaaggtagt atttgttggc gatcccccta gagtctttta catcttcgga aaacaaaaac
      421 tattttttct ttaatttctt tttttacttt ctatttttaa tttatatatt tatattaaaa
      481 aatttaaatt ataattattt ttatagcacg tgatgaaaag gacccaggtg gcgcggccgc
      541 caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
      601 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
      661 taggagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagg taactcacat
      721 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
      781 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
      841 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      901 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      961 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1021 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1081 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1141 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1201 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1261 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1321 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1381 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1441 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1501 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1561 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1621 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1681 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     1741 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     1801 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1861 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1921 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1981 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2041 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2101 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2161 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2221 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2281 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2341 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2401 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2461 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2521 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2581 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2641 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2701 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     2761 acgtctacaa ctcattacgc acactcatgg tttttggcca gcgaaaacag tttcaaaaga
     2821 ttgctggaag tctgcaataa tgtcatcaat aaattcgata ccaacagaga cacgaattaa
     2881 gtccttggta acaccagatg ccaacttttc tttgtcattt aattgtttgt gggtagtgaa
     2941 gtatggagca atgactaagg tcttggcatc accaacattg gccaagttag aggcaagctt
     3001 taaattgtca acaacttgag caccagaaag tttgaatggg tcagtttcct tgtcggcatt
     3061 tggtaagtct tttacaccga aagataagac accaccgaaa ccgttagata gatacttctt
     3121 agcattttca tgatgagaat gagatgctaa accagggtat gaaacccaag atacgtatgg
     3181 ggattgttct aaccatttgg ctaacttcaa tgcattttca ccgtgtcttt cagctctcaa
     3241 agataatgtt tcaacacctt gtagtagcaa gaaagaggca aatgggttca tcaatggacc
     3301 caaatctctt aatagttcag ttctaacatg aacgatgtat gccaagttac cgtaggcttc
     3361 attgtagata gtaccgtgat atccttcggc aggttgagag aattgaggga acttttctgg
     3421 gtagtccttc catgggaact taccagagtc aacaataata ccaccgatag tagtaccatg
     3481 accaccaatc catttggtag cagaatgtgt tacaatatca gcaccgtatt taattggctg
     3541 acagaagtaa ccaccggcac caaatgtgtt gtcaacgaca actggaatac cgtgtttgtg
     3601 agcaattgca acaatttttt caaaatccgg aacattgtac tttggattac caatggtttc
     3661 caaataaaca gccttggttc tttcatcaaa gaccttttcg aattcttctg gattgtcacc
     3721 ttcaacaaat ctagcctcga taccaaatct tttgaacgag attttgaact ggttataagt
     3781 accaccgtat aagtaagaag tggaaacgat gttgtcacca gtgtgtgcca aaccttggat
     3841 ggcaagggtt tgagcggctt gaccggagga aacagccaaa gcagcagcac caccttctaa
     3901 agcagcaatt ctttcttcca aaacattact ggttgggttt tggaaacggg aatagacgta
     3961 acctggaact tctagaccaa acaattgcga accatgctta gagttttcga aaacataaga
     4021 agtggtggcg taaattggta cagctctgga tctgtgagca ttgtcaccag ggttctcttg
     4081 gccggcgtgt agttgaacag tatcgaaatg agatggcatt gtatggatgg gggtaataga
     4141 attgtatcta tgtatctgac gaccctgtat tacacgaagg aaatagtaga tgaaaagaca
     4201 agagagcaag aaaaaggaaa aacttccttt ctaacagacg ctttacttaa ccttatatat
     4261 atcttatttt tttcatcgag cgtgttcaat tggacaaggt gccacctttt cgacacttcg
     4321 gttattatgt tacacagttt tcatgaggat ggctatcatg acattaacct ataaaaatag
     4381 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     4441 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     4501 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     4561 agagcagatt gtactgagag tgcaccatag atctgaatta attcttgaat aatacataac
     4621 ttttcttaaa agaatcaaag acagataaaa tttaagagat attaaatatt agtgagaagc
     4681 cgagaatttt gtaacaccaa cataacactg acatctttaa caacttttaa ttatgataca
     4741 tttcttacgt catgattgat tattacagct atgctgacaa atgactcttg ttgcatggct
     4801 acgaaccggg taatactaag tgattgactc ttgctgacct tttattaaga actaaatgga
     4861 caatattatg gagcatttca tgtataaatt ggtgcgtaaa atcgttggat ctctcttcta
     4921 agtacatcct actataacaa tcaagaaaaa caagaaaatc ggacaaaaca atcaagtatg
     4981 gattctagaa cagttggtat attaggaggg ggacaattgg gacgtatgat tgttgaggca
     5041 gcaaacaggc tcaacattaa gacggtaata ctagatgctg aaaattctcc tgccaaacaa
     5101 ataagcaact ccaatgacca cgttaatggc tccttttcca atcctcttga tatcgaaaaa
     5161 ctagctgaaa aatgtgatgt gctaacgatt gagattgagc atgttgatgt tcctacacta
     5221 aagaatcttc aagtaaaaca tcccaaatta aaaatttacc cttctccaga aacaatcaga
     5281 ttgatacaag acaaatatat tcaaaaagag catttaatca aaaatggtat agcagttacc
     5341 caaagtgttc ctgtggaaca agccagtgag acgtccctat tgaatgttgg aagagatttg
     5401 ggttttccat tcgtcttgaa gtcgaggact ttggcatacg atggaagagg taacttcgtt
     5461 gtaaagaata aggaaatgat tccggaagct ttggaagtac tgaaggatcg tcctttgtac
     5521 gccgaaaaat gggcaccatt tactaaagaa ttagcagtca tgattgtgag gtctgttaac
     5581 ggtttagtgt tttcttaccc aattgtagag actatccaca aggacaatat ttgtgactta
     5641 tgttatgcgc ctgctagagt tccggactcc gttcaactta aggcgaagtt gttggcagaa
     5701 aatgcaatca aatcttttcc cggttgtggt atatttggtg tggaaatgtt ctatttagaa
     5761 acaggggaat tgcttattaa cgaaattgcc ccaaggcctc acaactctgg acattatacc
     5821 attgatgctt gcgtcacttc tcaatttgaa gctcatttga gatcaatatt ggatttgcca
     5881 atgccaaaga atttcacatc tttctccacc attacaacga acgccattat gctaaatgtt
     5941 cttggagaca aacatacaaa agataaagag ctagaaactt gcgaaagagc attggcgact
     6001 ccaggttcct cagtgtactt atatggaaaa gagtctagac ctaacagaaa agtaggtcac
     6061 ataaatatta ttgcctccag tatggcggaa tgtgaacaaa ggctgaacta cattacaggt
     6121 agaactgata ttccaatcaa aatctctgtc gctcaaaagt tggacttgga agcaatggtc
     6181 aaaccattgg ttggaatcat catgggatca gactctgact tgccggtaat gtctgccgca
     6241 tgtgcggttt taaaagattt tggcgttcca tttgaagtga caatagtctc tgctcataga
     6301 actccacata ggatgtcagc atatgctatt tccgcaagca agcgtggaat taaaacaatt
     6361 atcgctggag ctggtggggc tgctcacttg ccaggtatgg tggctgcaat gacaccactt
     6421 cctgtcatcg gtgtgcccgt aaaaggttct tgtctagatg gagtagattc tttacattca
     6481 attgtgcaaa tgcctagagg tgttccagta gctaccgtcg ctattaataa tagtacgaac
     6541 gctgcgctgt tggctgtcag actgcttggc gcttatgatt caagttatac aacgaaaatg
     6601 gaacagtttt tattaaagca agaagaagaa gttcttgtca aagcacaaaa gttagaaact
     6661 gtcggttacg aagcttatct agaaaacaag taatatataa gtttattgat atacttgtac
     6721 agcaaataat tataaaatga tatacctatt ttttaggctt tgttatgatt acatcaaatg
     6781 tggacttcat acatagaaat caacgcttac aggtgtcctt ttttaagaat ttcatacata
     6841 agatctatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa
     6901 ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     6961 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     7021 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     7081 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     7141 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     7201 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     7261 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     7321 ccgccgcgct taatgcgccg ctacagggcg cgtcgcgcca ttcgccattc aggctgcgca
     7381 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     7441 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     7501 aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg ggtaccgggc
     7561 cccccctcga ggtcgacggt atcgataagc ttgatatcga attcctgcag cccgggggat
     7621 ccactagttc tagagcggcc gccctcttag cgct
//
