LOCUS       sequence-208922-        6666 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 6666)
  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 6666)
  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-cerMET16
FEATURES             Location/Qualifiers
     source          1..6666
                     /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             1325..1366
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1376..1393
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1414..1661
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1430..1448)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1738..1755)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1909..2497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1989..2008)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2668..3459,3460..3528))
                     /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     3291..3310
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(3624..4427)
                     /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(4428..4643)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5244..6121
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6132..6645)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6327..6348)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6539..6558
                     /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 tatgaagacc tatcatttga ataatgatat
      541 aattgtcaca caagaacagt tggatcattg gaatgaacaa ctaatcaagc tggaaacgcc
      601 acaggagatt attgcatggt ctatcgtaac gtttcctcac cttttccaaa ccactgcatt
      661 tggtttgact ggcttggtta ctatcgatat gttgtcaaag ctatctgaaa aatactacat
      721 gccagaacta ttatttatag acactttgca ccatttccca caaactttaa cactaaaaaa
      781 cgagattgag aaaaaatact accagcctaa aaatcaaacc attcacgtat ataagccgga
      841 tggatgtgaa tcggaggcag attttgcctc gaaatacggg gatttcttat gggagaaaga
      901 tgatgacaag tacgattatc tggccaaagt ggaacctgca catcgtgcct acaaagagct
      961 acatataagt gctgtgttta ctggtagaag aaaatcacaa ggttctgccc gctcccaact
     1021 gtcgattatt gaaatagacg aacttaatgg aatcttaaaa ataaatccat tgatcaattg
     1081 gacgttcgag caggttaaac agtatataga tgcaaacaat gtaccataca acgaactttt
     1141 ggaccttgga tatagatcca ttggtgatta ccattccaca caacccgtca aggaaggtga
     1201 agatgagaga gcaggaagat ggaagggcaa ggccaagacc gagtgtggaa ttcatgaagc
     1261 cagccgattc gcgcaatttt taaagcaaga tgccaagggc gagcttcgag gtcacccatt
     1321 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca
     1381 tcaccatcac cattgagttt ctagagggcc gcatcatgta attagttatg tcacgcttac
     1441 attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag
     1501 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     1561 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     1621 tgcttgagaa ggttttggga cgctcgaagg ctttaatttg caagctgcgg ccctgcatta
     1681 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1741 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1801 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1861 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1921 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1981 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2041 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2101 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2161 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2221 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2281 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2341 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2401 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2461 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2521 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2581 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2641 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2701 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2761 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2821 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2881 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2941 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3001 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3061 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3121 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3181 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3241 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3301 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3361 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3421 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3481 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3541 tcaatgggta ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat
     3601 gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc
     3661 cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt
     3721 cctcttccaa caataataat gtcagatcct gtagagacca catcatccac ggttctatac
     3781 tgttgaccca atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa
     3841 tcgtaacctt catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct
     3901 ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc
     3961 ttgcatgaca attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc
     4021 gcctgcttca aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct
     4081 gcccattctg ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg
     4141 tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc
     4201 ggcttaactg tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa
     4261 caaattttgg gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc
     4321 aatgaagcac acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca
     4381 ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt
     4441 ttcctgcagg tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct
     4501 tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc
     4561 ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa
     4621 taaaaaaaaa atgatgaatt gaaaagctag cttatcgatg ataagctgtc aaacatgaga
     4681 attaattcca cggactatag actataccta gtatactccg tctactgtac gatacacttc
     4741 cgctcaggtc cttgtccttt aacgaggcct taccactctt ttgttactct attgatccag
     4801 ctcagcaaag gcagtgtgat ctaagattct atcttcgcga tgtagtaaaa ctagctagac
     4861 cgagaaagag actagaaatg caaaaggcac ttctacaatg gctgccatca ttattatccg
     4921 atgtgacgct gcagcttctc aatgatattc gaatacgctt tgaggagata cagcctaata
     4981 tccgacaaac tgttttacag atttacgatc gtacttgtta cccatcattg aattttgaac
     5041 atccgaacct gggagttttc cctgaaacag atagtatatt tgaacctgta taataatata
     5101 tagtctagcg ctttacggaa gacaatgtat gtatttcggt tcctggagaa actattgcat
     5161 ctattgcata ggtaatcttg cacgtcgcat ccccggttca ttttctgcgt ttccatcttg
     5221 cacttcaata gcatatcttt gttaacgaag catctgtgct tcattttgta gaacaaaaat
     5281 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     5341 aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca
     5401 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     5461 acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt
     5521 ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt
     5581 tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta
     5641 aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca
     5701 cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat
     5761 ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc
     5821 gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat
     5881 actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc
     5941 ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt
     6001 cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag
     6061 cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa
     6121 tgggaagctc caccccggtt gataatcaga aaagccccaa aaacaggaag attgtataag
     6181 caaatattta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     6241 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     6301 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     6361 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     6421 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     6481 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     6541 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     6601 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtgggga tcgatccact
     6661 agtcca
//
