LOCUS       sequence-208926-        7344 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 7344)
  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 7344)
  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-cerMET2
FEATURES             Location/Qualifiers
     source          1..7344
                     /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             2003..2044
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2054..2071
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2092..2339
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2108..2126)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2416..2433)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2587..3175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2667..2686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3346..4137,4138..4206))
                     /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     3969..3988
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(4302..5105)
                     /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(5106..5321)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5922..6799
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6810..7323)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7005..7026)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7217..7236
                     /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 tggaatgtcg catactttaa aatcgaaaac
      541 gctccaagag ctggacattg aggagattaa ggaaactaac ccattgctca aactagttca
      601 agggcagagg attgttcaag ttccggaact agtgcttgag tctggcgtgg tcataaataa
      661 tttccctatt gcttataaga cgtggggtac actgaatgaa gctggtgata atgttctggt
      721 aatttgtcat gccttgactg ggtccacaga tgttgctgac tggtggggcc ctcttctggg
      781 taacgactta gcattcgacc catcaaggtt ttttatcata tgtttaaact ctatgggctc
      841 tccatatggg tctttttcgc cattaacgat aaatgaggag acgggcgtta gatatggacc
      901 cgaattccca ttatgtactg tgcgcgatga cgttagagct cacagaattg ttctggattc
      961 tctgggagta aagtcaatag cctgtgttat tggtggctct atggggggga tgctgagttt
     1021 ggaatgggct gccatgtatg gtaaggaata tgtgaagaat atggttgctc tggcgacatc
     1081 agcaagacat tctgcctggt gcatatcgtg gtctgaggct caaagacaat cgatttactc
     1141 agatcccaac tacttggacg ggtactatcc ggtagaggag caacctgtgg ccggactatc
     1201 ggctgcacgt atgtctgcat tgttgacgta caggacaaga aacagtttcg agaacaaatt
     1261 ctccagaaga tctccttcaa tagcacaaca acaaaaagct caaagggagg agacacgcaa
     1321 accatctact gtcagcgaac actccctaca aatccacaat gatgggtata aaacaaaagc
     1381 cagcactgcc atcgctggca tttctgagca aaaaggtcaa agcgtggtgt ccaccgcatc
     1441 ttcttcggat tcattgaatt cttcaacatc gatgacttcg gtaagttctg taacgggtga
     1501 agtgaaggac ataaagcctg cgcagacgta tttttctgca caaagttact tgaggtacca
     1561 gggcacaaag ttcatcaata ggttcgacgc caattgttac attgccatca cacgtaaact
     1621 ggatacgcac gatttggcaa gagacagagt agatgacatc actgaggtcc tttctaccat
     1681 ccaacaacca tccctgatca tcggtatcca atctgatgga ctgttcacat attcagaaca
     1741 agaatttttg gctgagcaca taccgaagtc gcaattagaa aaaattgaat ctcccgaagg
     1801 ccacgatgcc ttcctattgg agtttaagct gataaacaaa ctgatagtac aatttttaaa
     1861 aaccaactgc aaggccatta ccgatgccgc tccaagagct tggggaggtg acgttggtaa
     1921 cgatgaaacg aagacgtctg tctttggtga ggccgaagaa gttaccaact ggaagggcga
     1981 gcttcgaggt cacccattcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc
     2041 tacgcgtacc ggtcatcatc accatcacca ttgagtttct agagggccgc atcatgtaat
     2101 tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag
     2161 gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa
     2221 gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta
     2281 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgca
     2341 agctgcggcc ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2401 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2461 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2521 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2581 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2641 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2701 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2761 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2821 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2881 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2941 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3001 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3061 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3121 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3181 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3241 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3301 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3361 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3421 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3481 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3541 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3601 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3661 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3721 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3781 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3841 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3901 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3961 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4021 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4081 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4141 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4201 actcatactc ttcctttttc aatgggtaat aactgatata attaaattga agctctaatt
     4261 tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat
     4321 cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc
     4381 cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca
     4441 tcatccacgg ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg
     4501 ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata
     4561 aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct
     4621 ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc
     4681 tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg
     4741 tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat
     4801 ttgactgtat taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg
     4861 gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc
     4921 acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc
     4981 ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta
     5041 aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc
     5101 gacatgattt atcttcgttt cctgcaggtt tttgttctgt gcagttgggt taagaatact
     5161 gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc
     5221 tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac
     5281 cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga aaagctagct tatcgatgat
     5341 aagctgtcaa acatgagaat taattccacg gactatagac tatacctagt atactccgtc
     5401 tactgtacga tacacttccg ctcaggtcct tgtcctttaa cgaggcctta ccactctttt
     5461 gttactctat tgatccagct cagcaaaggc agtgtgatct aagattctat cttcgcgatg
     5521 tagtaaaact agctagaccg agaaagagac tagaaatgca aaaggcactt ctacaatggc
     5581 tgccatcatt attatccgat gtgacgctgc agcttctcaa tgatattcga atacgctttg
     5641 aggagataca gcctaatatc cgacaaactg ttttacagat ttacgatcgt acttgttacc
     5701 catcattgaa ttttgaacat ccgaacctgg gagttttccc tgaaacagat agtatatttg
     5761 aacctgtata ataatatata gtctagcgct ttacggaaga caatgtatgt atttcggttc
     5821 ctggagaaac tattgcatct attgcatagg taatcttgca cgtcgcatcc ccggttcatt
     5881 ttctgcgttt ccatcttgca cttcaatagc atatctttgt taacgaagca tctgtgcttc
     5941 attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     6001 gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg
     6061 cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     6121 gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat
     6181 ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc
     6241 atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt
     6301 tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat
     6361 aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt
     6421 ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg
     6481 tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct
     6541 tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat
     6601 tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa
     6661 cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta
     6721 ggttatatag ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg
     6781 tggaagcggt attcgcaatg ggaagctcca ccccggttga taatcagaaa agccccaaaa
     6841 acaggaagat tgtataagca aatatttaaa ttgtaagcgt taatattttg ttaaaattcg
     6901 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     6961 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     7021 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     7081 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     7141 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     7201 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     7261 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     7321 cgtggggatc gatccactag tcca
//
