LOCUS       sequence-208924-        7416 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of MET fusion protein in S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7416)
  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 7416)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "CBE Life 
            Sci Educ."; date: "2018-?"; volume: "17(1). pii: 17/1/ar1. doi"; 
            pages: " 10.1187/cbe.17-01-0012"
COMMENT     Sequence Label: pYES2.1-cerMET3
FEATURES             Location/Qualifiers
     source          1..7416
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             2075..2116
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2126..2143
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2164..2411
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2180..2198)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2488..2505)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2659..3247)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2739..2758)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3418..4209,4210..4278))
                     /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     4041..4060
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(4374..5177)
                     /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(5178..5393)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5994..6871
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6882..7395)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7077..7098)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7289..7308
                     /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 tatgcctgct cctcacggtg gtattctaca
      541 agacttgatt gctagagatg cgttaaagaa gaatgaattg ttatctgaag cgcaatcttc
      601 ggacatttta gtatggaact tgactcctag acaactatgt gatattgaat tgattctaaa
      661 tggtgggttt tctcctctga ctgggttttt gaacgaaaac gattactcct ctgttgttac
      721 agattcgaga ttagcagacg gcacattgtg gaccatccct attacattag atgttgatga
      781 agcatttgct aaccaaatta aaccagacac aagaattgcc cttttccaag atgatgaaat
      841 tcctattgct atacttactg tccaggatgt ttacaagcca aacaaaacta tcgaagccga
      901 aaaagtcttc agaggtgacc cagaacatcc agccattagc tatttattta acgttgccgg
      961 tgattattac gtcggcggtt ctttagaagc gattcaatta cctcaacatt atgactatcc
     1021 aggtttgcgt aagacacctg cccaactaag acttgaattc caatcaagac aatgggaccg
     1081 tgtcgtagct ttccaaactc gtaatccaat gcatagagcc cacagggagt tgactgtgag
     1141 agccgccaga gaagctaatg ctaaggtgct gatccatcca gttgttggac taaccaaacc
     1201 aggtgatata gaccatcaca ctcgtgttcg tgtctaccag gaaattatta agcgttatcc
     1261 taatggtatt gctttcttat ccctgttgcc attagcaatg agaatgagtg gtgatagaga
     1321 agccgtatgg catgctatta ttagaaagaa ttatggtgcc tcccacttca ttgttggtag
     1381 agaccatgcg ggcccaggta agaactccaa gggtgttgat ttctacggtc catacgatgc
     1441 tcaagaattg gtcgaatcct acaagcatga actggacatt gaagttgttc cattcagaat
     1501 ggtcacttat ttgccagacg aagaccgtta tgctccaatt gatcaaattg acaccacaaa
     1561 gacgagaacc ttgaacattt caggtacaga gttgagacgc cgtttaagag ttggtggtga
     1621 gattcctgaa tggttctcat atcctgaagt ggttaaaatc ctaagagaat ccaacccacc
     1681 aagaccaaaa caaggttttt caattgtttt aggtaattca ttaaccgttt ctcgtgagca
     1741 attatccatt gctttgttgt caacattctt gcaattcggt ggtggcaggt attacaagat
     1801 ctttgaacac aataataaga cagagttact atctttgatt caagatttca ttggttctgg
     1861 tagtggacta attattccaa atcaatggga agatgacaag gactctgttg ttggcaagca
     1921 aaacgtttac ttattagata cctcaagctc agccgatatt cagctagagt cagcggatga
     1981 acctatttca catattgtac aaaaagttgt cctattcttg gaagacaatg gcttttttgt
     2041 atttaagggc gagcttcgag gtcacccatt cgaaggtaag cctatcccta accctctcct
     2101 cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt ctagagggcc
     2161 gcatcatgta attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa
     2221 ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta
     2281 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg
     2341 tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg
     2401 ctttaatttg caagctgcgg ccctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2461 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2521 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2581 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2641 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2701 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2761 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2821 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2881 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2941 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3001 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3061 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3121 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3181 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3241 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3301 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3361 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3421 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3481 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3541 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3601 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3661 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3721 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3781 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3841 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3901 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3961 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4021 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4081 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4141 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4201 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4261 gaaatgttga atactcatac tcttcctttt tcaatgggta ataactgata taattaaatt
     4321 gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt tttttagttt
     4381 tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg ttcaccctct
     4441 accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat gtcagatcct
     4501 gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc cttgtcatct
     4561 aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc acccatgtct
     4621 ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc
     4681 ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa catcaaaagg
     4741 cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac aatacctggg
     4801 cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta tacacccgca
     4861 gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa
     4921 aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat ggaaaaatca
     4981 gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc ttcaactaac
     5041 tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt ttgcttttcg
     5101 tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc acgttcctta
     5161 tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg
     5221 gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat atataccaat
     5281 ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa
     5341 tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt gaaaagctag
     5401 cttatcgatg ataagctgtc aaacatgaga attaattcca cggactatag actataccta
     5461 gtatactccg tctactgtac gatacacttc cgctcaggtc cttgtccttt aacgaggcct
     5521 taccactctt ttgttactct attgatccag ctcagcaaag gcagtgtgat ctaagattct
     5581 atcttcgcga tgtagtaaaa ctagctagac cgagaaagag actagaaatg caaaaggcac
     5641 ttctacaatg gctgccatca ttattatccg atgtgacgct gcagcttctc aatgatattc
     5701 gaatacgctt tgaggagata cagcctaata tccgacaaac tgttttacag atttacgatc
     5761 gtacttgtta cccatcattg aattttgaac atccgaacct gggagttttc cctgaaacag
     5821 atagtatatt tgaacctgta taataatata tagtctagcg ctttacggaa gacaatgtat
     5881 gtatttcggt tcctggagaa actattgcat ctattgcata ggtaatcttg cacgtcgcat
     5941 ccccggttca ttttctgcgt ttccatcttg cacttcaata gcatatcttt gttaacgaag
     6001 catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     6061 agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac
     6121 gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca
     6181 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta
     6241 ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt
     6301 ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc
     6361 ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat
     6421 tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc
     6481 tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt
     6541 gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta
     6601 tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt
     6661 attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat
     6721 actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa
     6781 ggtggatggg taggttatat agggatatag cacagagata tatagcaaag agatactttt
     6841 gagcaatgtt tgtggaagcg gtattcgcaa tgggaagctc caccccggtt gataatcaga
     6901 aaagccccaa aaacaggaag attgtataag caaatattta aattgtaagc gttaatattt
     6961 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     7021 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     7081 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     7141 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     7201 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     7261 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     7321 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     7381 cgctacaggg cgcgtgggga tcgatccact agtcca
//
