LOCUS       sequence-207581-        6489 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 6489)
  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 6489)
  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-cerMET14
FEATURES             Location/Qualifiers
     source          1..6489
                     /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             1148..1189
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1199..1216
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1237..1484
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1253..1271)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1561..1578)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1732..2320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1812..1831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2491..3282,3283..3351))
                     /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     3114..3133
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(3447..4250)
                     /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(4251..4466)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5067..5944
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(5955..6468)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6150..6171)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6362..6381
                     /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 tatggctact aatattactt ggcatccaaa
      541 tcttacttac gacgaacgca aggcattgag aaaacaggac ggttgtacta tttggttaac
      601 aggtctaagt gcgtcaggta aaagtacaat cgcctgtgcg ctagaacagt tactgctcca
      661 aaaaaacttg tctgcatata gattggatgg tgacaacatt cgttttggat tgaacaagga
      721 tttgggtttc tcagaaaagg acagaaatga aaacattcgt agaattagcg aagtttctaa
      781 gctatttgct gattcatgtg ctatttcaat cacctcattt atctctccat acagagttga
      841 cagagataga gctcgtgaac tacataagga ggctggtttg aagttcattg aaatatttgt
      901 tgatgttcca ttagaagtcg ctgagcaaag ggaccctaag ggtttataca agaaagctag
      961 ggagggtgta atcaaggagt ttacaggtat ttctgcccca tatgaagcgc caaaagctcc
     1021 agagctacat ttgagaaccg accagaagac ggttgaagaa tgtgctacca ttatttatga
     1081 gtacttaatc agtgaaaaaa tcatccgtaa gcatttgaag ggcgagcttc gaggtcaccc
     1141 attcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca
     1201 tcatcaccat caccattgag tttctagagg gccgcatcat gtaattagtt atgtcacgct
     1261 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
     1321 aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat
     1381 ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa
     1441 ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcaagctg cggccctgca
     1501 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1561 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1621 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1681 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1741 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1801 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1861 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1921 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1981 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2041 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2101 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2161 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2221 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2281 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2341 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2401 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2461 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2521 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2581 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2641 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2701 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2761 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2821 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2881 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2941 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3001 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3061 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3121 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3181 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3241 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3301 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3361 ttttcaatgg gtaataactg atataattaa attgaagctc taatttgtga gtttagtata
     3421 catgcattta cttataatac agttttttag ttttgctggc cgcatcttct caaatatgct
     3481 tcccagcctg cttttctgta acgttcaccc tctaccttag catcccttcc ctttgcaaat
     3541 agtcctcttc caacaataat aatgtcagat cctgtagaga ccacatcatc cacggttcta
     3601 tactgttgac ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt cataatcaac
     3661 caatcgtaac cttcatctct tccacccatg tctctttgag caataaagcc gataacaaaa
     3721 tctttgtcgc tcttcgcaat gtcaacagta cccttagtat attctccagt agatagggag
     3781 cccttgcatg acaattctgc taacatcaaa aggcctctag gttcctttgt tacttcttct
     3841 gccgcctgct tcaaaccgct aacaatacct gggcccacca caccgtgtgc attcgtaatg
     3901 tctgcccatt ctgctattct gtatacaccc gcagagtact gcaatttgac tgtattacca
     3961 atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt acttggcgga taatgccttt
     4021 agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga tatccacatg tgtttttagt
     4081 aaacaaattt tgggacctaa tgcttcaact aactccagta attccttggt ggtacgaaca
     4141 tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga tattaaatag cttggcagca
     4201 acaggactag gatgagtagc agcacgttcc ttatatgtag ctttcgacat gatttatctt
     4261 cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga atactgggca atttcatgtt
     4321 tcttcaacac tacatatgcg tatatatacc aatctaagtc tgtgctcctt ccttcgttct
     4381 tccttctgtt cggagattac cgaatcaaaa aaatttcaag gaaaccgaaa tcaaaaaaaa
     4441 gaataaaaaa aaaatgatga attgaaaagc tagcttatcg atgataagct gtcaaacatg
     4501 agaattaatt ccacggacta tagactatac ctagtatact ccgtctactg tacgatacac
     4561 ttccgctcag gtccttgtcc tttaacgagg ccttaccact cttttgttac tctattgatc
     4621 cagctcagca aaggcagtgt gatctaagat tctatcttcg cgatgtagta aaactagcta
     4681 gaccgagaaa gagactagaa atgcaaaagg cacttctaca atggctgcca tcattattat
     4741 ccgatgtgac gctgcagctt ctcaatgata ttcgaatacg ctttgaggag atacagccta
     4801 atatccgaca aactgtttta cagatttacg atcgtacttg ttacccatca ttgaattttg
     4861 aacatccgaa cctgggagtt ttccctgaaa cagatagtat atttgaacct gtataataat
     4921 atatagtcta gcgctttacg gaagacaatg tatgtatttc ggttcctgga gaaactattg
     4981 catctattgc ataggtaatc ttgcacgtcg catccccggt tcattttctg cgtttccatc
     5041 ttgcacttca atagcatatc tttgttaacg aagcatctgt gcttcatttt gtagaacaaa
     5101 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     5161 agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa
     5221 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     5281 agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt
     5341 gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt
     5401 ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg
     5461 ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact
     5521 ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg
     5581 catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat
     5641 agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta
     5701 tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag
     5761 ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga
     5821 ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata
     5881 tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg
     5941 caatgggaag ctccaccccg gttgataatc agaaaagccc caaaaacagg aagattgtat
     6001 aagcaaatat ttaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     6061 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     6121 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     6181 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     6241 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     6301 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     6361 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     6421 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtgg ggatcgatcc
     6481 actagtcca
//
