LOCUS       sequence-208890-        6705 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 6705)
  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 6705)
  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-cerMET8
FEATURES             Location/Qualifiers
     source          1..6705
                     /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             1364..1405
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1415..1432
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1453..1700
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1469..1487)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1777..1794)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1948..2536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2028..2047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2707..3498,3499..3567))
                     /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     3330..3349
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(3663..4466)
                     /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(4467..4682)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5283..6160
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6171..6684)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6366..6387)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6578..6597
                     /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 tatggtcaaa tcgctacagc tagcccatca
      541 attaaaagac aagaaaatac tactaatcgg aggcggagaa gtcggcttga caaggttata
      601 taagctgata cctacaggtt gcaagctgac tttagtatct cctgacctac acaaatccat
      661 tattccgaag tttggaaaat tcatccagaa cgaggatcag cccgactaca gagaagacgc
      721 taaacgcttt atcaatccga actgggaccc cacgaagaat gaaatttacg agtacatccg
      781 cagtgacttc aaagacgaat acctcgacct agaagacgag aacgacgcgt ggtacataat
      841 aatgacctgt atacccgacc accctgaaag cgcaagaata tatcaccttt gtaaggaaag
      901 attcggtaag cagcagctgg tcaatgtggc ggacaaacct gatctctgtg acttttattt
      961 cggcgccaac ctggagatag gagaccgtct acagattctg atctctacca acggcctttc
     1021 tccgcgcttt ggcgctctag taagagatga gatccgtaac ttattcacac aaatggggga
     1081 tctggcgctg gaggacgctg tcgtcaaact aggtgagctg agaagaggaa ttaggctgct
     1141 ggcaccagac gacaaggatg tcaagtaccg catggattgg gccagacgct gcacagacct
     1201 cttcggcatt cagcactgcc acaacatcga cgtaaaacgt ctgctggatc ttttcaaggt
     1261 catgtttcaa gaacagaact gttccttgca gttccctccc agagaacggt tgcttagcga
     1321 gtactgctcg tctaagggcg agcttcgagg tcacccattc gaaggtaagc ctatccctaa
     1381 ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc attgagtttc
     1441 tagagggccg catcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat
     1501 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt
     1561 ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
     1621 acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac
     1681 gctcgaaggc tttaatttgc aagctgcggc cctgcattaa tgaatcggcc aacgcgcggg
     1741 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     1801 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     1861 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     1921 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     1981 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2041 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2101 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2161 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2221 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2281 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2341 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2401 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2461 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2521 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2581 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2641 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2701 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     2761 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     2821 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     2881 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     2941 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3001 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3061 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3121 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3181 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3241 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3301 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3361 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3421 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3481 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3541 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatgggtaa taactgatat
     3601 aattaaattg aagctctaat ttgtgagttt agtatacatg catttactta taatacagtt
     3661 ttttagtttt gctggccgca tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt
     3721 tcaccctcta ccttagcatc ccttcccttt gcaaatagtc ctcttccaac aataataatg
     3781 tcagatcctg tagagaccac atcatccacg gttctatact gttgacccaa tgcgtctccc
     3841 ttgtcatcta aacccacacc gggtgtcata atcaaccaat cgtaaccttc atctcttcca
     3901 cccatgtctc tttgagcaat aaagccgata acaaaatctt tgtcgctctt cgcaatgtca
     3961 acagtaccct tagtatattc tccagtagat agggagccct tgcatgacaa ttctgctaac
     4021 atcaaaaggc ctctaggttc ctttgttact tcttctgccg cctgcttcaa accgctaaca
     4081 atacctgggc ccaccacacc gtgtgcattc gtaatgtctg cccattctgc tattctgtat
     4141 acacccgcag agtactgcaa tttgactgta ttaccaatgt cagcaaattt tctgtcttcg
     4201 aagagtaaaa aattgtactt ggcggataat gcctttagcg gcttaactgt gccctccatg
     4261 gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac aaattttggg acctaatgct
     4321 tcaactaact ccagtaattc cttggtggta cgaacatcca atgaagcaca caagtttgtt
     4381 tgcttttcgt gcatgatatt aaatagcttg gcagcaacag gactaggatg agtagcagca
     4441 cgttccttat atgtagcttt cgacatgatt tatcttcgtt tcctgcaggt ttttgttctg
     4501 tgcagttggg ttaagaatac tgggcaattt catgtttctt caacactaca tatgcgtata
     4561 tataccaatc taagtctgtg ctccttcctt cgttcttcct tctgttcgga gattaccgaa
     4621 tcaaaaaaat ttcaaggaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg
     4681 aaaagctagc ttatcgatga taagctgtca aacatgagaa ttaattccac ggactataga
     4741 ctatacctag tatactccgt ctactgtacg atacacttcc gctcaggtcc ttgtccttta
     4801 acgaggcctt accactcttt tgttactcta ttgatccagc tcagcaaagg cagtgtgatc
     4861 taagattcta tcttcgcgat gtagtaaaac tagctagacc gagaaagaga ctagaaatgc
     4921 aaaaggcact tctacaatgg ctgccatcat tattatccga tgtgacgctg cagcttctca
     4981 atgatattcg aatacgcttt gaggagatac agcctaatat ccgacaaact gttttacaga
     5041 tttacgatcg tacttgttac ccatcattga attttgaaca tccgaacctg ggagttttcc
     5101 ctgaaacaga tagtatattt gaacctgtat aataatatat agtctagcgc tttacggaag
     5161 acaatgtatg tatttcggtt cctggagaaa ctattgcatc tattgcatag gtaatcttgc
     5221 acgtcgcatc cccggttcat tttctgcgtt tccatcttgc acttcaatag catatctttg
     5281 ttaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt
     5341 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat
     5401 tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct
     5461 aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc
     5521 gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga
     5581 gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa
     5641 tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt
     5701 ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac
     5761 tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg
     5821 atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc
     5881 agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta
     5941 cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta
     6001 aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa
     6061 ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga
     6121 gatacttttg agcaatgttt gtggaagcgg tattcgcaat gggaagctcc accccggttg
     6181 ataatcagaa aagccccaaa aacaggaaga ttgtataagc aaatatttaa attgtaagcg
     6241 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     6301 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     6361 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     6421 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     6481 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     6541 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     6601 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     6661 ttaatgcgcc gctacagggc gcgtggggat cgatccacta gtcca
//
