LOCUS       sequence-207590-        8988 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 8988)
  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 8988)
  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-cerMET10
FEATURES             Location/Qualifiers
     source          1..8988
                     /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             3647..3688
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3698..3715
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3736..3983
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3752..3770)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4060..4077)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4231..4819)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4311..4330)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4990..5781,5782..5850))
                     /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     5613..5632
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(5946..6749)
                     /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(6750..6965)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      7566..8443
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(8454..8967)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8649..8670)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8861..8880
                     /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 tatgccagtt gagtttgcta ccaatccttt
      541 tggcgaggcc aaaaatgcaa cttcactgcc aaaatatggt acacccgtaa ctgccatttc
      601 atctgtgctg ttcaataacg tggactccat ttttgcttac aagtcctttt ctcaacccga
      661 tttgctacac caagatctaa aaaaatggtc tgaaaagcgt ggtaacgaat cacgtgggaa
      721 gccatttttc caagagctgg atatcagatc tggcgctggt ttggctcctt tagggttttc
      781 tcatggattg aagaacacta cagcaattgt tgctccaggg ttttcgctgc catacttcat
      841 taactctttg aaaaccgtct ctcatgatgg taagtttctt ttgaatgttg gtgctttaaa
      901 ctacgacaat gctaccggct ctgtcaccaa cgattatgta accgcattgg atgctgcttc
      961 caagctgaag tatggtgtcg tgactccgat ttccgctaac gaggtacaaa gtgtcgcctt
     1021 actggcattg gcgattgcca ctttcagtaa taactccgga gctatcaatt tatttgacgg
     1081 attaaactac tcgaaaaccg tcttgccgtt ggtcgaatct gttcctgagg catctatttt
     1141 ggcaaaacta tccaaagtta ttgcaccaga tgctgccttt gatgatgtct tggataagtt
     1201 taatgaattg actggattga gactacataa tttccaatac tttggtgctc aggatgctga
     1261 aactgtgttt atcacttatg ggtctttaga atccgaattg ttcaactctg cgattagtgg
     1321 taataattcc aaaatcgggt taatcaacgt cagagtgcca ttacctttta acgttgctaa
     1381 gtttgtcact cacgttccat ccactaccaa acaaattgtt gttataggcc aaactttgga
     1441 tggttcttcg ccttctttct tgagatctca agtttcagcc gccttatttt accacggccg
     1501 cacctcaatt agcgtttctg agtacatcta tcaaccagat ttcatttggt ccccaaaagc
     1561 tgtcaaatca attgtatcgt cattcatccc tgaattcact tacaatgccg attcatcttt
     1621 cggcgaagga ttcatttatt gggcctctga taagagtatc aatattgatg ttgcctccaa
     1681 acttgtgaaa gctctgtctt tggaagatgg gaaatttgtg tctttgagaa cgaaatttga
     1741 taacttggct aatgctggta ccttccaagc tcaatttgtg acctcgaaag aacagatacc
     1801 tgtttcaaac atcgattcta cgaaattatc agtcgttgaa gatgtcagtt tattgaagca
     1861 tttagacgta gctgctaccg tcgcagaaca aggttcaatt gcgttggttt cccaaaaggc
     1921 agttaaagat ttggatttaa attctgtaga aagttacgtc aagaatttgg gaattcctga
     1981 atcattccta atatctattg cgaagaaaaa catcaaattg tttatcatcg atggtgagac
     2041 cactaacgac gagtccaaat tgtccttgtt tatccaagcc gttttctgga aattggcctt
     2101 cggtctagat gtcgcagaat gtaccaaccg tatctggaaa agcattgatt caggtgcaga
     2161 catttcagca gcctcgattt ctgaatttct cactggtgca ttcaaaaact tcctcagtga
     2221 ggttccgcta gcgctgtaca ctaaattttc tgaaataaac attgaaaaga aagaggatga
     2281 ggaagagcct gcagctttac caattttcgt taatgaaaca tctttcctcc caaataacag
     2341 taccattgaa gaaataccat tacctgagac ctctgagatc tctgatattg ccaagaagtt
     2401 gtccttcaaa gaggcatatg aagttgagaa taaactaaga cccgatttac ccgtcaagaa
     2461 cttcgtcgtg aaagttaaag aaaatagacg tgttacgcct gctgattatg atagatatat
     2521 tttccatatt gaattcgata tttctggtac tggaatgact tatgacatcg gtgaagccct
     2581 cggtattcat gccagaaaca atgaatcttt ggtcaaagaa ttcttaacct tctatggtct
     2641 aaatgaatcc gatgttgtct tagtccccaa caaggacaac caccatttgt tagaaacaag
     2701 aaccgtctta caagcatttg tggaaaattt ggatattttc ggtaaaccac caaaaagatt
     2761 ttacgaatca ttgattccat atgcctctaa cgaagaggag aagaaaaaat tagaggattt
     2821 ggttactcct gccggtgcag tagatttgaa gagatttcaa gatgtggagt attatacata
     2881 tgctgacatt tttgaattgt tcccatctgt tcgcccatct cttgaggaac ttgttactat
     2941 cattgaacca ttgaagagaa gagaatactc aattgcctcc tctcagaaag ttcatccaaa
     3001 tgaagttcat ttattgatcg ttgttgttga ttgggtggat aataaaggaa gaaaaaggta
     3061 cggtcaagct tctaagtata tctcagacct tgctgtcggt tcagaattgg tcgttagcgt
     3121 taaaccatct gttatgaaat taccaccatc tccaaagcaa ccagttatta tgagtggttt
     3181 aggtactggt ttggcaccat tcaaggccat tgttgaagag aaattatggc aaaagcagca
     3241 aggttatgag attggtgaag tcttcctata tctaggttca agacacaaaa gagaagaata
     3301 tttatatggt gagttatggg aggcttacaa agatgcaggt attatcacac acatcggcgc
     3361 tgctttctca agagaccaac ctcaaaaaat ttacattcaa gatcgtatca aagagaattt
     3421 ggatgaatta aaaactgcaa tgattgataa taaaggttca ttttacttgt gtggccctac
     3481 ttggccagtt ccagatatta ctcaagcttt gcaagacatt ctggctaaag acgccgagga
     3541 aagaggcatc aaagtcgact tggatgccgc aattgaagaa ttaaaggaag catcaagata
     3601 cattttagaa gtctacaagg gcgagcttcg aggtcaccca ttcgaaggta agcctatccc
     3661 taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt
     3721 ttctagaggg ccgcatcatg taattagtta tgtcacgctt acattcacgc cctcccccca
     3781 catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt
     3841 tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc
     3901 tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg
     3961 gacgctcgaa ggctttaatt tgcaagctgc ggccctgcat taatgaatcg gccaacgcgc
     4021 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4081 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4141 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4201 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4261 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4321 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4381 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4441 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4501 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4561 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4621 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4681 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4741 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4801 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4861 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4921 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4981 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5041 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5101 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5161 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5221 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5281 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5341 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5401 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5461 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5521 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5581 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5641 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5701 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5761 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5821 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatggg taataactga
     5881 tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca
     5941 gttttttagt tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa
     6001 cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata
     6061 atgtcagatc ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct
     6121 cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt
     6181 ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg
     6241 tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct
     6301 aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta
     6361 acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg
     6421 tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct
     6481 tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc
     6541 atggaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat
     6601 gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt
     6661 gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca
     6721 gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca ggtttttgtt
     6781 ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt
     6841 atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc
     6901 gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa
     6961 ttgaaaagct agcttatcga tgataagctg tcaaacatga gaattaattc cacggactat
     7021 agactatacc tagtatactc cgtctactgt acgatacact tccgctcagg tccttgtcct
     7081 ttaacgaggc cttaccactc ttttgttact ctattgatcc agctcagcaa aggcagtgtg
     7141 atctaagatt ctatcttcgc gatgtagtaa aactagctag accgagaaag agactagaaa
     7201 tgcaaaaggc acttctacaa tggctgccat cattattatc cgatgtgacg ctgcagcttc
     7261 tcaatgatat tcgaatacgc tttgaggaga tacagcctaa tatccgacaa actgttttac
     7321 agatttacga tcgtacttgt tacccatcat tgaattttga acatccgaac ctgggagttt
     7381 tccctgaaac agatagtata tttgaacctg tataataata tatagtctag cgctttacgg
     7441 aagacaatgt atgtatttcg gttcctggag aaactattgc atctattgca taggtaatct
     7501 tgcacgtcgc atccccggtt cattttctgc gtttccatct tgcacttcaa tagcatatct
     7561 ttgttaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa
     7621 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc
     7681 tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc
     7741 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag
     7801 agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg
     7861 agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta
     7921 taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac
     7981 tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat
     8041 tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt atattctata
     8101 ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg
     8161 gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt
     8221 ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt
     8281 ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt
     8341 caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa
     8401 agagatactt ttgagcaatg tttgtggaag cggtattcgc aatgggaagc tccaccccgg
     8461 ttgataatca gaaaagcccc aaaaacagga agattgtata agcaaatatt taaattgtaa
     8521 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     8581 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     8641 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     8701 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     8761 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     8821 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     8881 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     8941 cgcttaatgc gccgctacag ggcgcgtggg gatcgatcca ctagtcca
//
