LOCUS       sequence-208846-        7368 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 7368)
  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 7368)
  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-pomMet1
FEATURES             Location/Qualifiers
     source          1..7368
                     /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             2027..2068
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2078..2095
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2116..2363
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2132..2150)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2440..2457)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2611..3199)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2691..2710)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3370..4161,4162..4230))
                     /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     3993..4012
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(4326..5129)
                     /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(5130..5345)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5946..6823
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6834..7347)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7029..7050)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7241..7260
                     /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 tatgattgtt tcctggtcct tgaaagacgc
      541 cgttgttgta gtcgttggtt caggagaaaa tgcttaccga cgggtgaatt tatgtttaag
      601 tgaaaattct tctaaaattt tttggttttg tcgcagctcc gctgatggtg gatttgacgt
      661 tggtcatctt cagcttgacg caaaaaaatc ggatgctatt gaaattcttc ccatttctga
      721 atttgatccc gttcgttcat taacgacgct gggaaaggag gagacagatt ttttggtgga
      781 tgctgttttc gtgtcagagt ccaaccacca tgaaaaggaa attttacatc gtacgtgcaa
      841 aaggtaccga gtcccattaa acatcattga taatccttct ttgtgctcct ttactctacc
      901 agctacgtgg tcagaaccac ctcttcaaat ttctttaagt acttcatcaa atggttgtcg
      961 tttagctcaa aggctgttga gacacgtggt gagttgcctt ccttccggta tgccagaagc
     1021 cattgagcgt tttggccgag tgcgagcgat cactaaaact ccagaaaaaa aacagtggat
     1081 caatcatgtt agtgattttt ggcccctcga gaagttggtt cggatgacag aggacgatct
     1141 tcttgccata gtttctgaca acttctctct tccactctca tcaagccaat cttcttcact
     1201 tgccaactcc tatgaaagtt tatcaaccac cttggacaag ccttcattga cgcttgaccc
     1261 tgaggctttt cctactcata agcgaggttc aatagcttta attggaagtg gacctggctc
     1321 tcctgattta cttactgttg ctgccagaaa ggctataatg aaagccgatt atgtactagc
     1381 cgacaagctg gtaccagaag cggtattaca gttaattcca aggcatactc cactatttat
     1441 tgctagaaaa tttcctggaa atgcggataa ggcacaagat gaacttcacc aggttgcttt
     1501 tgatgcttta tctagaggtg attatgttgt ccgtttgaag caaggtgatc cttatattta
     1561 tgggcgaggc ggtgaagagt atctattctt tacacaacac ggttatgtac caacggttat
     1621 tcctggcatc agtagtgctt tgatggcccc tatctctgct ggtattcccg ttacacatcg
     1681 tggagtggcc gatcaatttc ttgtgtgtac tggaactggt caaaagggtt cgatgcctaa
     1741 aataccttct tttgtcccta cacagaccac agttttccta atggctttac accgactaga
     1801 aattcttgtg caagcactaa ttgaaagcgg ttggcctaga gtgttaccgg tttgtatagc
     1861 tgagcgcgtc tcttgccctg atcaaaggtt cattttctct actttggaag acgttgtgga
     1921 agaatacaac aagtacgagt ctctcccccc tggtttgctg attactggat acagttgtaa
     1981 tacccttcgc aacaccaagg gcgagcttcg aggtcaccca ttcgaaggta agcctatccc
     2041 taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt
     2101 ttctagaggg ccgcatcatg taattagtta tgtcacgctt acattcacgc cctcccccca
     2161 catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt
     2221 tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc
     2281 tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg
     2341 gacgctcgaa ggctttaatt tgcaagctgc ggccctgcat taatgaatcg gccaacgcgc
     2401 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2461 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2521 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2581 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2641 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2701 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2761 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2821 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2881 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2941 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3001 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3061 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3121 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3181 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3241 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3301 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3361 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3421 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3481 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3541 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3601 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3661 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3721 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3781 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3841 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3901 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3961 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4021 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4081 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4141 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4201 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatggg taataactga
     4261 tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca
     4321 gttttttagt tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa
     4381 cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata
     4441 atgtcagatc ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct
     4501 cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt
     4561 ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg
     4621 tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct
     4681 aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta
     4741 acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg
     4801 tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct
     4861 tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc
     4921 atggaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat
     4981 gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt
     5041 gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca
     5101 gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca ggtttttgtt
     5161 ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt
     5221 atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc
     5281 gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa
     5341 ttgaaaagct agcttatcga tgataagctg tcaaacatga gaattaattc cacggactat
     5401 agactatacc tagtatactc cgtctactgt acgatacact tccgctcagg tccttgtcct
     5461 ttaacgaggc cttaccactc ttttgttact ctattgatcc agctcagcaa aggcagtgtg
     5521 atctaagatt ctatcttcgc gatgtagtaa aactagctag accgagaaag agactagaaa
     5581 tgcaaaaggc acttctacaa tggctgccat cattattatc cgatgtgacg ctgcagcttc
     5641 tcaatgatat tcgaatacgc tttgaggaga tacagcctaa tatccgacaa actgttttac
     5701 agatttacga tcgtacttgt tacccatcat tgaattttga acatccgaac ctgggagttt
     5761 tccctgaaac agatagtata tttgaacctg tataataata tatagtctag cgctttacgg
     5821 aagacaatgt atgtatttcg gttcctggag aaactattgc atctattgca taggtaatct
     5881 tgcacgtcgc atccccggtt cattttctgc gtttccatct tgcacttcaa tagcatatct
     5941 ttgttaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa
     6001 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc
     6061 tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc
     6121 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag
     6181 agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg
     6241 agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta
     6301 taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac
     6361 tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat
     6421 tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt atattctata
     6481 ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg
     6541 gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt
     6601 ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt
     6661 ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt
     6721 caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa
     6781 agagatactt ttgagcaatg tttgtggaag cggtattcgc aatgggaagc tccaccccgg
     6841 ttgataatca gaaaagcccc aaaaacagga agattgtata agcaaatatt taaattgtaa
     6901 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     6961 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     7021 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     7081 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     7141 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     7201 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     7261 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     7321 cgcttaatgc gccgctacag ggcgcgtggg gatcgatcca ctagtcca
//
