LOCUS       Exported                7215 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Overexpression of MET fusion protein in S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    pYES2.1-cerMET17
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7215)
  AUTHORS   Reeves TD, Warner DM, Ludlow LH, O'Connor CM
  TITLE     Pathways over Time: Functional Genomics Research in an Introductory 
            Laboratory Course.
  JOURNAL   CBE Life Sci Educ. 2018 Spring;17(1). pii: 17/1/ar1. doi: 
            10.1187/cbe.17-01-0012.
  PUBMED    29326101
REFERENCE   2  (bases 1 to 7215)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7215
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..442
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1..118
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     413..436
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     474..493
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        474..492
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             512..1843
                     /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"
     CDS             1874..1915
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1925..1942
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1963..2210
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1979..1997)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2287..2304)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2458..3046)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2538..2557)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3217..4077)
                     /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"
     primer_bind     3840..3859
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(4173..4976)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4977..5192)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5793..6670
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6681..7194)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6876..6897)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7088..7107
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cggattagaa gccgccgagc gggtgacagc cctccgaagg aagactctcc tccgtgcgtc
       61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
      121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
      181 tggccccaca aaccttcaaa tgaacgaatc aaattaacaa ccataggatg ataatgcgat
      241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
      301 aacagatata taaatgcaaa aactgcataa ccactttaac taatactttc aacattttcg
      361 gtttgtatta cttcttattc aaatgtaata aaagtatcaa caaaaaattg ttaatatacc
      421 tctatacttt aacgtcaagg agaaaaaacc ccggatcgga ctactagcag ctgtaatacg
      481 actcactata gggaatatta agctcgccct tatgccatct catttcgata ctgttcaact
      541 acacgccggc caagagaacc ctggtgacaa tgctcacaga tccagagctg taccaattta
      601 cgccaccact tcttatgttt tcgaaaactc taagcatggt tcgcaattgt ttggtctaga
      661 agttccaggt tacgtctatt cccgtttcca aaacccaacc agtaatgttt tggaagaaag
      721 aattgctgct ttagaaggtg gtgctgctgc tttggctgtt tcctccggtc aagccgctca
      781 aacccttgcc atccaaggtt tggcacacac tggtgacaac atcgtttcca cttcttactt
      841 atacggtggt acttataacc agttcaaaat ctcgttcaaa agatttggta tcgaggctag
      901 atttgttgaa ggtgacaatc cagaagaatt cgaaaaggtc tttgatgaaa gaaccaaggc
      961 tgtttatttg gaaaccattg gtaatccaaa gtacaatgtt ccggattttg aaaaaattgt
     1021 tgcaattgct cacaaacacg gtattccagt tgtcgttgac aacacatttg gtgccggtgg
     1081 ttacttctgt cagccaatta aatacggtgc tgatattgta acacattctg ctaccaaatg
     1141 gattggtggt catggtacta ctatcggtgg tattattgtt gactctggta agttcccatg
     1201 gaaggactac ccagaaaagt tccctcaatt ctctcaacct gccgaaggat atcacggtac
     1261 tatctacaat gaagcctacg gtaacttggc atacatcgtt catgttagaa ctgaactatt
     1321 aagagatttg ggtccattga tgaacccatt tgcctctttc ttgctactac aaggtgttga
     1381 aacattatct ttgagagctg aaagacacgg tgaaaatgca ttgaagttag ccaaatggtt
     1441 agaacaatcc ccatacgtat cttgggtttc ataccctggt ttagcatctc attctcatca
     1501 tgaaaatgct aagaagtatc tatctaacgg tttcggtggt gtcttatctt tcggtgtaaa
     1561 agacttacca aatgccgaca aggaaactga cccattcaaa ctttctggtg ctcaagttgt
     1621 tgacaattta aagctcgcct ctaacttggc caatgttggt gatgccaaga ccttagtcat
     1681 tgctccatac ttcactaccc acaaacaatt aaatgacaaa gaaaagttgg catctggtgt
     1741 taccaaggac ttaattcgtg tctctgttgg tatcgaattt attgatgaca ttattgcaga
     1801 cttccagcaa tcttttgaaa ctgttttcgc tggccaaaaa ccaaagggcg agcttcgagg
     1861 tcacccattc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     1921 cggtcatcat caccatcacc attgagtttc tagagggccg catcatgtaa ttagttatgt
     1981 cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac
     2041 aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat
     2101 ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac
     2161 tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc aagctgcggc
     2221 cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2281 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2341 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2401 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2461 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2521 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2581 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2641 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2701 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2761 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2821 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2881 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2941 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3001 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3061 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3121 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3181 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3241 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3301 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3361 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3421 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3481 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3541 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3601 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3661 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3721 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3781 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3841 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3901 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3961 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4021 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4081 cttccttttt caatgggtaa taactgatat aattaaattg aagctctaat ttgtgagttt
     4141 agtatacatg catttactta taatacagtt ttttagtttt gctggccgca tcttctcaaa
     4201 tatgcttccc agcctgcttt tctgtaacgt tcaccctcta ccttagcatc ccttcccttt
     4261 gcaaatagtc ctcttccaac aataataatg tcagatcctg tagagaccac atcatccacg
     4321 gttctatact gttgacccaa tgcgtctccc ttgtcatcta aacccacacc gggtgtcata
     4381 atcaaccaat cgtaaccttc atctcttcca cccatgtctc tttgagcaat aaagccgata
     4441 acaaaatctt tgtcgctctt cgcaatgtca acagtaccct tagtatattc tccagtagat
     4501 agggagccct tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc ctttgttact
     4561 tcttctgccg cctgcttcaa accgctaaca atacctgggc ccaccacacc gtgtgcattc
     4621 gtaatgtctg cccattctgc tattctgtat acacccgcag agtactgcaa tttgactgta
     4681 ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa aattgtactt ggcggataat
     4741 gcctttagcg gcttaactgt gccctccatg gaaaaatcag tcaagatatc cacatgtgtt
     4801 tttagtaaac aaattttggg acctaatgct tcaactaact ccagtaattc cttggtggta
     4861 cgaacatcca atgaagcaca caagtttgtt tgcttttcgt gcatgatatt aaatagcttg
     4921 gcagcaacag gactaggatg agtagcagca cgttccttat atgtagcttt cgacatgatt
     4981 tatcttcgtt tcctgcaggt ttttgttctg tgcagttggg ttaagaatac tgggcaattt
     5041 catgtttctt caacactaca tatgcgtata tataccaatc taagtctgtg ctccttcctt
     5101 cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat ttcaaggaaa ccgaaatcaa
     5161 aaaaaagaat aaaaaaaaaa tgatgaattg aaaagctagc ttatcgatga taagctgtca
     5221 aacatgagaa ttaattccac ggactataga ctatacctag tatactccgt ctactgtacg
     5281 atacacttcc gctcaggtcc ttgtccttta acgaggcctt accactcttt tgttactcta
     5341 ttgatccagc tcagcaaagg cagtgtgatc taagattcta tcttcgcgat gtagtaaaac
     5401 tagctagacc gagaaagaga ctagaaatgc aaaaggcact tctacaatgg ctgccatcat
     5461 tattatccga tgtgacgctg cagcttctca atgatattcg aatacgcttt gaggagatac
     5521 agcctaatat ccgacaaact gttttacaga tttacgatcg tacttgttac ccatcattga
     5581 attttgaaca tccgaacctg ggagttttcc ctgaaacaga tagtatattt gaacctgtat
     5641 aataatatat agtctagcgc tttacggaag acaatgtatg tatttcggtt cctggagaaa
     5701 ctattgcatc tattgcatag gtaatcttgc acgtcgcatc cccggttcat tttctgcgtt
     5761 tccatcttgc acttcaatag catatctttg ttaacgaagc atctgtgctt cattttgtag
     5821 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     5881 cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt
     5941 tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     6001 ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc
     6061 ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat
     6121 tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta
     6181 ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc
     6241 ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga
     6301 taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa
     6361 agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg
     6421 tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat
     6481 gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa
     6541 tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata
     6601 gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg
     6661 tattcgcaat gggaagctcc accccggttg ataatcagaa aagccccaaa aacaggaaga
     6721 ttgtataagc aaatatttaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     6781 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     6841 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     6901 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     6961 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     7021 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     7081 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     7141 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtggggat
     7201 cgatccacta gtcca
//
