LOCUS       sequence-208923-        8184 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 8184)
  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 8184)
  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-cerMET6
FEATURES             Location/Qualifiers
     source          1..8184
                     /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             2843..2884
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2894..2911
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2932..3179
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2948..2966)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3256..3273)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3427..4015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3507..3526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4186..4977,4978..5046))
                     /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     4809..4828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(5142..5945)
                     /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(5946..6161)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      6762..7639
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(7650..8163)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7845..7866)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8057..8076
                     /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 tatggttcaa tctgctgtct tagggttccc
      541 aagaatcggt ccaaacagag aattaaagaa ggccactgaa ggttactgga acggtaaaat
      601 cactgtcgat gaattattca aggtcggtaa ggatttgaga actcaaaact ggaagttgca
      661 aaaggaggct ggtgttgata tcatcccatc caatgacttc tccttttacg accaagtttt
      721 ggatttgtct ttgttgttca atgtcattcc agaccgttac actaagtacg atctatctcc
      781 aatcgacact ttgtttgcta tgggtagagg tttacaaaga aaggccactg aaactgaaaa
      841 ggctgtcgac gtcactgctt tggaaatggt taaatggttc gactctaact accattacgt
      901 tagaccaact ttctccaaga ccactcaatt taagttgaac ggccaaaagc cagttgacga
      961 atttttggaa gccaaggagt taggtattca tactagacct gtcttgttgg gtccagtttc
     1021 ttacttattc ttgggtaagg ctgacaagga ttctctagat ttggaaccat tgtccctatt
     1081 ggaacaattg ttgcctctat acactgaaat cctatctaaa ttggcttctg ctggtgccac
     1141 tgaagttcaa attgacgaac ctgtcttagt tttggacttg cctgccaacg cccaagccgc
     1201 cattaagaag gcttacactt acttcggtga acaaagcaat ctaccaaaga ttactttggc
     1261 tacttacttc ggtaccgttg tccctaactt agacgccatc aagggcttgc cagttgctgc
     1321 cttacacgtt gactttgtta gagctccaga acaatttgat gaagtcgttg ccgccattgg
     1381 taacaaacaa accttgtccg ttggtattgt tgatggtaga aacatttgga agaatgattt
     1441 caagaagtct tccgctatcg ttaacaaggc tattgaaaag ttgggtgctg acagagtcgt
     1501 tgttgccact tcttcttctc tattgcacac accagttgat ttgaacaacg aaaccaagtt
     1561 ggacgctgaa atcaagggct ttttctcttt cgccactcaa aaattggatg aagttgttgt
     1621 gatcaccaag aacgtttccg gtcaagacgt tgctgctgcc ctagaagcta acgctaaatc
     1681 tgttgaatcc agaggtaaat ccaagtttat ccacgatgct gccgttaagg ccagagttgc
     1741 ctctatcgac gaaaaaatgt ctactagagc agctccattt gaacaaagat tgcctgaaca
     1801 acaaaaagtc ttcaacttgc cattgttccc aacaacaact attggttcct tccctcaaac
     1861 caaggacatc agaattaaca gaaacaaatt caacaagggc accatctctg ctgaagaata
     1921 tgaaaaattc atcaattctg aaattgaaaa ggtcatcaga ttccaagaag aaattggttt
     1981 ggatgtctta gtccacggtg aaccagaaag aaacgatatg gttcaatact tcggtgaaca
     2041 aatcaacggt tatgctttca ctgttaacgg ttgggttcaa tcttacggtt ccagatatgt
     2101 cagaccacca attattgttg gtgacttgtc cagaccaaag gctatgtccg tcaaggaatc
     2161 tgtttacgct caatccatca cttctaagcc agtaaagggt atgttgactg gtccaattac
     2221 ctgtttgaga tggtctttcc caagagacga tgtcgaccaa aaaactcaag ctatgcaatt
     2281 agctttggct ttgagagatg aagtcaatga tttggaagct gccggtatca aggttatcca
     2341 agttgatgaa ccagctttaa gagaaggttt accattgaga gaaggtactg agagatctgc
     2401 ttactacacc tgggctgccg aagctttcag agttgctact tctggtgttg ctaacaagac
     2461 tcaaatacac tctcatttct gttactctga cttggatcca aaccatatca aggctttgga
     2521 tgctgatgtt gtttccatcg aattctctaa gaaggacgat gctaactaca ttgctgaatt
     2581 caaaaactat ccaaaccaca ttggtctagg tttattcgat attcattctc caagaattcc
     2641 atcaaaggat gaatttatcg ccaagatttc aaccatcttg aagagctacc cagctgaaaa
     2701 gttctgggtt aacccagatt gtggtttgaa gactagaggc tgggaagaaa ctagattgtc
     2761 tttgactcat atggtcgaag ccgccaagta cttccgtgaa caatacaaga ataagggcga
     2821 gcttcgaggt cacccattcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc
     2881 tacgcgtacc ggtcatcatc accatcacca ttgagtttct agagggccgc atcatgtaat
     2941 tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag
     3001 gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa
     3061 gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta
     3121 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgca
     3181 agctgcggcc ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3241 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3301 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3361 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3421 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3481 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3541 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3601 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3661 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3721 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3781 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3841 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3901 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3961 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4261 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4381 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5041 actcatactc ttcctttttc aatgggtaat aactgatata attaaattga agctctaatt
     5101 tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat
     5161 cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc
     5221 cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca
     5281 tcatccacgg ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg
     5341 ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata
     5401 aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct
     5461 ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc
     5521 tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg
     5581 tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat
     5641 ttgactgtat taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg
     5701 gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc
     5761 acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc
     5821 ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta
     5881 aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc
     5941 gacatgattt atcttcgttt cctgcaggtt tttgttctgt gcagttgggt taagaatact
     6001 gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc
     6061 tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac
     6121 cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga aaagctagct tatcgatgat
     6181 aagctgtcaa acatgagaat taattccacg gactatagac tatacctagt atactccgtc
     6241 tactgtacga tacacttccg ctcaggtcct tgtcctttaa cgaggcctta ccactctttt
     6301 gttactctat tgatccagct cagcaaaggc agtgtgatct aagattctat cttcgcgatg
     6361 tagtaaaact agctagaccg agaaagagac tagaaatgca aaaggcactt ctacaatggc
     6421 tgccatcatt attatccgat gtgacgctgc agcttctcaa tgatattcga atacgctttg
     6481 aggagataca gcctaatatc cgacaaactg ttttacagat ttacgatcgt acttgttacc
     6541 catcattgaa ttttgaacat ccgaacctgg gagttttccc tgaaacagat agtatatttg
     6601 aacctgtata ataatatata gtctagcgct ttacggaaga caatgtatgt atttcggttc
     6661 ctggagaaac tattgcatct attgcatagg taatcttgca cgtcgcatcc ccggttcatt
     6721 ttctgcgttt ccatcttgca cttcaatagc atatctttgt taacgaagca tctgtgcttc
     6781 attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     6841 gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg
     6901 cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     6961 gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat
     7021 ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc
     7081 atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt
     7141 tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat
     7201 aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt
     7261 ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg
     7321 tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct
     7381 tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat
     7441 tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa
     7501 cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta
     7561 ggttatatag ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg
     7621 tggaagcggt attcgcaatg ggaagctcca ccccggttga taatcagaaa agccccaaaa
     7681 acaggaagat tgtataagca aatatttaaa ttgtaagcgt taatattttg ttaaaattcg
     7741 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     7801 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     7861 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     7921 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     7981 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     8041 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     8101 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     8161 cgtggggatc gatccactag tcca
//
