LOCUS       sequence-208876-        7170 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 7170)
  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 7170)
  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-pomMet17
FEATURES             Location/Qualifiers
     source          1..7170
                     /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"
     regulatory      637..646
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1829..1870
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1880..1897
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1918..2165
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1934..1952)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2242..2259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2413..3001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2493..2512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3172..3963,3964..4032))
                     /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     3795..3814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(4128..4931)
                     /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(4932..5147)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5748..6625
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6636..7149)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6831..6852)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7043..7062
                     /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 tatgccagtc gagagtgaac atttcgaaac
      541 tttacaatta catgctggcc aagagcctga tgctgctacc agctctcgtg ccgttcccat
      601 ctacgctact acttcctatg ttttccgtga ttgcgaccat ggcggccgct tgttcggatt
      661 acaggaacca ggttacatct actcgcgtat gatgaatccc accgccgacg tttttgagaa
      721 acgtattgcc gccttggagc atggcgctgc tgcaatcgct actagttccg gtacttccgc
      781 tctcttcatg gctttgacca cgttggctaa ggccggtgat aacattgtct ccacttctta
      841 cctttatggt ggtacttaca acctcttcaa ggttaccctg cctagattgg gaattactac
      901 caagtttgtc aatggtgatg atcctaatga tcttgcagct cagattgatg aaaacacaaa
      961 ggctgtttac gttgagtcca tcggcaatcc catgtacaac gttcccgatt ttgagcgtat
     1021 cgctgaggtt gctcatgccg ctggtgtgcc tttaatggtc gataacactt ttggcggcgg
     1081 tggttatttg gttcgtccca ttgaccacgg tgccgatatc gttacccact ctgccactaa
     1141 gtggatcggt ggtcatggca ctactattgg cggtgtgatt gttgatagtg gtaagtttga
     1201 ctggaagaag aacagcaagc gtttccctga attcaacgag cctcatcccg gttaccatgg
     1261 catggtcttt actgaaactt ttggtaactt ggcatatgct tttgcttgcc gtactcaaac
     1321 tctccgtgat gttggtggca atgccaatcc attcggtgtc tttttgcttc ttcaaggtct
     1381 tgaaacgctt tctcttcgta tggagcgtca cgttcaaaat gcatttgctc ttgcaaaata
     1441 tttggaaaag caccccaagg ttaactgggt ttcttaccct ggtcttgaat ctcacgtctc
     1501 tcacaaactt gccaagaagt acttgaaaaa tggttacggc gccgttctca gctttggcgc
     1561 taaaggtggc cctgatcaaa gtcgtaaggt agtcaatgcc ttaaagcttg ctagtcagtt
     1621 ggccaatgtt ggtgatgcca aaactttggt tatcgctcct gcctatacca ctcatttaca
     1681 attaactgat gaggagcaaa tttctgccgg tgtcactaag gatcttattc gtgtggccgt
     1741 cggtattgag cacatcgatg atattatcgc cgactttgct caagctttgg aagttgccaa
     1801 gggcgagctt cgaggtcacc cattcgaagg taagcctatc cctaaccctc tcctcggtct
     1861 cgattctacg cgtaccggtc atcatcacca tcaccattga gtttctagag ggccgcatca
     1921 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa
     1981 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag
     2041 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg
     2101 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa
     2161 tttgcaagct gcggccctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2221 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2281 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2341 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2401 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2461 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2521 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2581 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2641 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2701 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2761 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2821 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2881 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2941 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3001 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3061 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3121 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3181 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3241 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3301 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3361 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3421 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3481 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3541 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3601 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3661 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3721 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3781 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3841 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3901 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3961 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4021 ttgaatactc atactcttcc tttttcaatg ggtaataact gatataatta aattgaagct
     4081 ctaatttgtg agtttagtat acatgcattt acttataata cagtttttta gttttgctgg
     4141 ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta
     4201 gcatcccttc cctttgcaaa tagtcctctt ccaacaataa taatgtcaga tcctgtagag
     4261 accacatcat ccacggttct atactgttga cccaatgcgt ctcccttgtc atctaaaccc
     4321 acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc ttccacccat gtctctttga
     4381 gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt acccttagta
     4441 tattctccag tagataggga gcccttgcat gacaattctg ctaacatcaa aaggcctcta
     4501 ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc
     4561 acaccgtgtg cattcgtaat gtctgcccat tctgctattc tgtatacacc cgcagagtac
     4621 tgcaatttga ctgtattacc aatgtcagca aattttctgt cttcgaagag taaaaaattg
     4681 tacttggcgg ataatgcctt tagcggctta actgtgccct ccatggaaaa atcagtcaag
     4741 atatccacat gtgtttttag taaacaaatt ttgggaccta atgcttcaac taactccagt
     4801 aattccttgg tggtacgaac atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg
     4861 atattaaata gcttggcagc aacaggacta ggatgagtag cagcacgttc cttatatgta
     4921 gctttcgaca tgatttatct tcgtttcctg caggtttttg ttctgtgcag ttgggttaag
     4981 aatactgggc aatttcatgt ttcttcaaca ctacatatgc gtatatatac caatctaagt
     5041 ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa
     5101 ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag ctagcttatc
     5161 gatgataagc tgtcaaacat gagaattaat tccacggact atagactata cctagtatac
     5221 tccgtctact gtacgataca cttccgctca ggtccttgtc ctttaacgag gccttaccac
     5281 tcttttgtta ctctattgat ccagctcagc aaaggcagtg tgatctaaga ttctatcttc
     5341 gcgatgtagt aaaactagct agaccgagaa agagactaga aatgcaaaag gcacttctac
     5401 aatggctgcc atcattatta tccgatgtga cgctgcagct tctcaatgat attcgaatac
     5461 gctttgagga gatacagcct aatatccgac aaactgtttt acagatttac gatcgtactt
     5521 gttacccatc attgaatttt gaacatccga acctgggagt tttccctgaa acagatagta
     5581 tatttgaacc tgtataataa tatatagtct agcgctttac ggaagacaat gtatgtattt
     5641 cggttcctgg agaaactatt gcatctattg cataggtaat cttgcacgtc gcatccccgg
     5701 ttcattttct gcgtttccat cttgcacttc aatagcatat ctttgttaac gaagcatctg
     5761 tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc
     5821 tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac caacgaagaa
     5881 tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa
     5941 gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat tttaccaaca
     6001 aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct atttttctaa
     6061 caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag tctcttgata
     6121 actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt ctattttctc
     6181 ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg aagctgcggg
     6241 tgcatttttt caagataaag gcatccccga ttatattcta taccgatgtg gattgcgcat
     6301 actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa attatgaacg
     6361 gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt tcgtattgtt
     6421 ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag taatactaga
     6481 gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga
     6541 tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac ttttgagcaa
     6601 tgtttgtgga agcggtattc gcaatgggaa gctccacccc ggttgataat cagaaaagcc
     6661 ccaaaaacag gaagattgta taagcaaata tttaaattgt aagcgttaat attttgttaa
     6721 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     6781 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     6841 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     6901 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     6961 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     7021 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     7081 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     7141 agggcgcgtg gggatcgatc cactagtcca
//
