LOCUS       sequence-208869-        8172 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 8172)
  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 8172)
  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-pomMet26
FEATURES             Location/Qualifiers
     source          1..8172
                     /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             2831..2872
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2882..2899
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2920..3167
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2936..2954)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3244..3261)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3415..4003)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3495..3514)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4174..4965,4966..5034))
                     /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     4797..4816
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(5130..5933)
                     /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(5934..6149)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      6750..7627
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(7638..8151)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7833..7854)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8045..8064
                     /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 tatggttaaa tctgctgttt tggggttccc
      541 tcgtatcggt aagaaccgtg agttgaagaa ggccaccgag gcttactggt ctggaaagac
      601 cagcgccgag gaattgcttg ctactgccaa gcaattgcgt ttggagcact ggaagcttca
      661 aaaggcccaa ggtgttgaca ttatcccttc caatgatttc tcattgtatg atcaaatcat
      721 ggatcactct ttctctttca atgttatccc ccccagatac cgcctctccg gtttgtcttc
      781 tttggatact tactttgcca tgggccgtgg tatgcaacgt gctgctactg ccgataaggc
      841 tgccgttgat gtccccgccg gtgagatggt caagtggttt gacagcaact accatttctt
      901 gcgtcccgag gtttctgagg agactgactt caagttgagc tctactaagg ctctcgacga
      961 gttccttgag gctaaggaag ctggtatcat cactcgccct gttcttgttg gtcccgttac
     1021 ctacttgttc attgctaagg ccgccaaggg ttctagcatt aagcctattg aactcttgcc
     1081 caagctttta cccgtttatg ttgagcttat caagaagctt actgaggctg gtgcagagta
     1141 catccaaatt gatgagccta tccttactct tgacttgccc caagagatcc ttgctagcta
     1201 caaggaagct tatgagactc ttggtaagat cggtaagctc attttgacca cctacttcgg
     1261 ctctcttcaa agcaatgccg atgttttgaa gggtctccct attgctggtg tccacgttga
     1321 tgtcgttcgt gctcctgaga acttggaccg tgctcttgcc gttctcggtg agaaccaaat
     1381 catctctgtt ggtgtcgttt ccggaagaaa catctggaag actgatttcc aaaaggccac
     1441 tgccatcatt gagaaggcta ttagcgctgt cggcagtgag cgtgtgcaag ttgcttcttc
     1501 ctccagcatt cttcacattc ctcactccct ttctggcgaa gaccaaatta atcctgaaat
     1561 taagcgttgg ttcgcctttg ctgttgaaaa gtgtgctgaa ttagctattc ttactaaggc
     1621 tgctaatgac ggccctgcct ccgttcgtgc tgaactcgag gccaacgctg ctgactgcaa
     1681 agctcgtgct gagtctccca tcaccaatgt tgaggccgtt cgtgaacgtc aaagcaaggt
     1741 tactccccaa atgcatgaac gtaagagtcc ctttgagacc cgttatgcca agcaacaagc
     1801 ttcccttaag cttcctcttt tccctaccac cactattggc tcttttcctc aaaccaagga
     1861 aattcgtgta accagaaaca gattcgctaa gggtctcatt tctcaagagg aatatgatgc
     1921 cttcatccgt aaggagattt ctgatgttgt caaattccaa gaggaagttg gtcttgacgt
     1981 tctggttcac ggtgaacccg agcgtaacga tatggttcaa tactttggtg aaagaatgga
     2041 gggctttgtc tttactgtca acggttgggt ccaatcttat ggttcccgtt gtgtccgtcc
     2101 tcctatcatt gtcggtgatg tttatcgtcc cgcccctatg accgttaagg agtctcaata
     2161 tgctcaatcc attacttcta agcctatgaa gggtatgttg actgctccca tcactattct
     2221 tagatggtct ttccctcgtg acgatgttca tgactctgtc caagctcaac aaattgctct
     2281 tggtctcaga gatgaggtcc ttgatcttga aaaggctggt attaaagtca ttcaatgtga
     2341 tgagcctgcc ttacgtgaag gtcttcctct tcgtcgtgcc gagtgggatg agtatctcaa
     2401 atgggctatc gatgctttcc gtttggctac tgctgctgtt caagatgaca ctcaaattca
     2461 ctcccatttc tgttattctg acttcaatga cattttcgat gccattcaac gtttggatgc
     2521 tgacgttgtt tccattgaga actccaagtc tgacatgaag ttgttgaatg ttcttagccg
     2581 ctatacctct tgcatcggtc ccggtctttt cgatatccac tctcctcgtg ttcctcctgt
     2641 ttccgagttc aaggagcgta ttgatgctat tgtcaagcat gttcctaagg atcatctctg
     2701 gctcaaccct gactgtggtt tgaagacccg tggatggcct gagactactg ctgacttgaa
     2761 gaacatgatt gccgctgctc gtgaagctcg tgagcaatac aagggcgagc ttcgaggtca
     2821 cccattcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg
     2881 tcatcatcac catcaccatt gagtttctag agggccgcat catgtaatta gttatgtcac
     2941 gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac
     3001 ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta
     3061 tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga
     3121 aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag ctgcggccct
     3181 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3241 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3301 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3361 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3421 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3481 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3541 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3601 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3661 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3721 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3781 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3841 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3901 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3961 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4021 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4081 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4141 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4201 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4261 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4321 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4381 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4441 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4501 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4561 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4621 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4681 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4741 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4801 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4861 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4921 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4981 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5041 cctttttcaa tgggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt
     5101 atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat
     5161 gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca
     5221 aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc atccacggtt
     5281 ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg tgtcataatc
     5341 aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca
     5401 aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg
     5461 gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct
     5521 tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta
     5581 atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta
     5641 ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc
     5701 tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt
     5761 agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga
     5821 acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca
     5881 gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat
     5941 cttcgtttcc tgcaggtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat
     6001 gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt
     6061 tcttccttct gttcggagat taccgaatca aaaaaatttc aaggaaaccg aaatcaaaaa
     6121 aaagaataaa aaaaaaatga tgaattgaaa agctagctta tcgatgataa gctgtcaaac
     6181 atgagaatta attccacgga ctatagacta tacctagtat actccgtcta ctgtacgata
     6241 cacttccgct caggtccttg tcctttaacg aggccttacc actcttttgt tactctattg
     6301 atccagctca gcaaaggcag tgtgatctaa gattctatct tcgcgatgta gtaaaactag
     6361 ctagaccgag aaagagacta gaaatgcaaa aggcacttct acaatggctg ccatcattat
     6421 tatccgatgt gacgctgcag cttctcaatg atattcgaat acgctttgag gagatacagc
     6481 ctaatatccg acaaactgtt ttacagattt acgatcgtac ttgttaccca tcattgaatt
     6541 ttgaacatcc gaacctggga gttttccctg aaacagatag tatatttgaa cctgtataat
     6601 aatatatagt ctagcgcttt acggaagaca atgtatgtat ttcggttcct ggagaaacta
     6661 ttgcatctat tgcataggta atcttgcacg tcgcatcccc ggttcatttt ctgcgtttcc
     6721 atcttgcact tcaatagcat atctttgtta acgaagcatc tgtgcttcat tttgtagaac
     6781 aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag
     6841 aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt
     6901 aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     6961 tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt
     7021 tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac
     7081 tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt
     7141 ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg
     7201 actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa
     7261 aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt
     7321 gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct
     7381 ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa
     7441 tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt
     7501 agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg
     7561 atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat
     7621 tcgcaatggg aagctccacc ccggttgata atcagaaaag ccccaaaaac aggaagattg
     7681 tataagcaaa tatttaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     7741 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     7801 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     7861 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     7921 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     7981 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     8041 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     8101 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tggggatcga
     8161 tccactagtc ca
//
