LOCUS       sequence-202677-       10305 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 10305)
  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 10305)
  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-pomSir1
FEATURES             Location/Qualifiers
     source          1..10305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             53..94
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             104..121
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      142..389
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(158..176)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(466..483)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(637..1225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(717..736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1396..2187,2188..2256))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLID*MEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(2352..3155)
                     /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(3156..3376)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      3975..4855
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(4924..5379)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     5273..5292
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5398..5839
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    5398..5515
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     5810..5833
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
ORIGIN
        1 agggaatatt aagctcgccc ttaagggcga gcttcgaggt cacccattcg aaggtaagcc
       61 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca
      121 ttgagtttct agagggccgc atcatgtaat tagttatgtc acgcttacat tcacgccctc
      181 cccccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta
      241 tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt
      301 tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg
      361 ttttgggacg ctcgaaggct ttaatttgca agctgcggcc ctgcattaat gaatcggcca
      421 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
      481 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      541 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      601 gcccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      661 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      721 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      781 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      841 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      901 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      961 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1021 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     1081 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1141 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1201 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1261 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1321 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1381 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1441 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggagcg
     1501 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1561 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1621 atccgcctcc attcagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1681 taatagtttg cgcaacgttg ttggcattgc tacaggcatc gtggtgtcac tctcgtcgtt
     1741 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1801 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1861 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1921 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1981 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aatagtgtat cacatagcag
     2041 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2101 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2161 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2221 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatgggtaat
     2281 aactgatata attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat
     2341 aatacagttt tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt
     2401 ctgtaacgtt caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca
     2461 ataataatgt cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat
     2521 gcgtctccct tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca
     2581 tctcttccac ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc
     2641 gcaatgtcaa cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat
     2701 tctgctaaca tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa
     2761 ccgctaacaa tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct
     2821 attctgtata cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt
     2881 ctgtcttcga agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg
     2941 ccctccatgg aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga
     3001 cctaatgctt caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac
     3061 aagtttgttt gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga
     3121 gtagcagcac gttccttata tgtagctttc gacatgattt atcttcgttt cctgcaggtt
     3181 tttgttctgt gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat
     3241 atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag
     3301 attaccgaat caaaaaaatt tcaaagaaac cgaaatcaaa aaaaagaata aaaaaaaaat
     3361 gatgaattga attgaaaagc tagcttatcg atgataagct gtcaaagatg agaattaatt
     3421 ccacggacta tagactatac tagatactcc gtctactgta cgatacactt ccgctcaggt
     3481 ccttgtcctt taacgaggcc ttaccactct tttgttactc tattgatcca gctcagcaaa
     3541 ggcagtgtga tctaagattc tatcttcgcg atgtagtaaa actagctaga ccgagaaaga
     3601 gactagaaat gcaaaaggca cttctacaat ggctgccatc attattatcc gatgtgacgc
     3661 tgcagcttct caatgatatt cgaatacgct ttgaggagat acagcctaat atccgacaaa
     3721 ctgttttaca gatttacgat cgtacttgtt acccatcatt gaattttgaa catccgaacc
     3781 tgggagtttt ccctgaaaca gatagtatat ttgaacctgt ataataatat atagtctagc
     3841 gctttacgga agacaatgta tgtatttcgg ttcctggaga aactattgca tctattgcat
     3901 aggtaatctt gcacgtcgca tccccggttc attttctgcg tttccatctt gcacttcaat
     3961 agcatatctt tgttaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga
     4021 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     4081 cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa
     4141 cgcgacgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     4201 atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa
     4261 aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct
     4321 ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta
     4381 gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc
     4441 gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga
     4501 ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat
     4561 gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg
     4621 tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta
     4681 caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt
     4741 tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga
     4801 gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatgggaa
     4861 gctccacccc ggttgataat cagaaaagcc ccaaaaacag gaagattgta taagcaaata
     4921 tttaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     4981 cattttttaa cgaatagccc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     5041 agatagggtt gagtgttgtt ccagtttcca acaagagtcc actattaaag aacgtggact
     5101 ccaacgtcaa agggcgaaaa agggtctatc agggcgatgg cccactacgt gaaccatcac
     5161 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcagt aaatcggaag ggtaaacgga
     5221 tgcccccatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     5281 aagcgaaagg agcgggggct agggcggtgg gaagtgtagg ggtcacgctg ggcgtaacca
     5341 ccacacccgc cgcgcttaat ggggcgctac agggcgcgtg gggatgatcc actagtacgg
     5401 attagaagcc gccgagcggg tgacagccct ccgaaggaag actctcctcc gtgcgtcctc
     5461 gtcttcaccg gtcgcgttcc tgaaacgcag atgtgcctcg cgccgcactg ctccgaacaa
     5521 taaagattct acaatactag cttttatggt tatgaagagg aaaaattggc agtaacctgg
     5581 ccccacaaac cttcaaatga acgaatcaaa ttaacaacca taggatgata atgcgattag
     5641 ttttttagcc ttatttctgg ggtaattaat cagcgaagcg atgatttttg atctattaac
     5701 agatatataa atgcaaaaac tgcataacca ctttaactaa tactttcaac attttcggtt
     5761 tgtattactt cttattcaaa tgtaataaaa gtatcaacaa aaaattgtta atatacctct
     5821 atactttaac gtcaaggaga aaaaaccccg gatcggacta ctagcagctg taatacgact
     5881 cactatatga gtgtaaaggc tagtatcaat aattctcagg aggctgtctc acgcatcgct
     5941 ttccgttgct cagatgcgtt gtacgtggtt catcctaaca attcgatttt aaatggtgct
     6001 ttaacagaat ccttaaaaga tttaaaaaaa tttgaaacgt taaatgtttc cggcaaggtt
     6061 ccacacgttc ttcctttaaa atctcacgct gatccctttg ctcacattgc ggatgctatt
     6121 ttggctgaag aagaagtggc ttctactcaa cctaaacaaa tcacatctgt agttgcttct
     6181 gctgatgcac tctttttcgc tactcctcat ctttataagt tggctcatga gcctcttgta
     6241 gcgcacgttg ccatagagtc tacagaggca tttgattttg cttctgtgcg tgacaccggt
     6301 ttcgttattc tcttttcggg caatcgccct ggtgattctt cggaggctgc tttggaagat
     6361 accttggaga cagcttcttt ggctcatcgc ttagcattga aattgaacac aggcgtttta
     6421 catttctatt ctccagttta cgacacgacc gctgcgttgg agaacatcga gacactacct
     6481 tccaaagagg atgctcaaca cgctagagtt gcccacattc ctattgaaga gaaacaggag
     6541 gattctgaaa aagaaggcaa tatcaaggaa gcttttgtcc ctcccaagtt tgatcagcca
     6601 gaacgtgatg ccgctacttc tgaatacctt gaatcattgt ctatcaagcc ttttgaatac
     6661 tctggttccg atgacgctac tgatgtcttg ttagttttcg gaagcgctgc ttctgaatta
     6721 gccaaggctg ccgttacttc ctctgtggca gttgctattg taagagtcct tcgcccttgg
     6781 ttgccctcta aattgcaaga agtgcttcct acttctacta agcgcttgac tgttttggaa
     6841 cccattacca gtcttcctcg taaatgggat ccgctttacc ttgacgtttt atcttctttc
     6901 gtcgcttccg gatcttcaat tgagctgttt gccgtaagat acggattgag ttcttcagag
     6961 caagccacag agattattaa agctgttcgt gataatcttt caggcgcttt aaagccatct
     7021 ctcgtttgcg attttaccga tggtgtttct caagtgtttg ttcctacacc tccgtctatt
     7081 gaggaagctt atcataaact tcttcataga gtctttaagt ctcgtcttaa cattgttaac
     7141 gatcccgctt catcagctac gaaacaaaac attccatctc gtctcatcat atctcctcag
     7201 tttgctttgg gttcggtttt agagtatgaa aatcagcgtc gcgctttctg tgatgaggtt
     7261 gctactttat tgaaagaaaa gaattcttct gtctcttctg aatcattaga agttctttcg
     7321 aattggattg tttccgttga taacctggag tctcctgttg accctgagtt ggtcatctct
     7381 gaacttaaga aggattcatc tgctcctatc aaatcacttt tagatcgatc tgaatttttt
     7441 accaacgttt cacattggat tatcggttcc gatgcttggg catatgactt gggtaattct
     7501 gctcttcatc aagtactttg cttggaaaag aacgtcaatt tattgattgt tgacacccag
     7561 ccttactcta ctcgtgaagc cgtccgatcc tcttccagaa agaaggatat tggattgtat
     7621 gccatgaact ttggcaatgc atatgtcgcc tctactgcgt tgtatagttc atacacccaa
     7681 ctaatctcgg cacttttaga agcagataaa tttaagggcc cctcggttgt tttggcatac
     7741 cttccctatc acagtgctga cgatgatgcc attactgtgt tacaggaaac caagaaagcc
     7801 gttgacatcg gttattggcc tttgtataga tggactcctg ctttagagga cggagaatat
     7861 tcagatttca aacttgattc cgaacgtatt cgccgagagc tcaagacctt tttggaaagg
     7921 gataactact tgactcaact aacgcttcgt gtgccaagtt tagctcgtac tcttactcaa
     7981 tcatttggtg cggaagtccg tcatcaacaa aacgttgact cacgcaatgc tctcaacaaa
     8041 ttaattgaag gtttgtctgg tcctcctctt accattcttt tcgcttcaga cggaggtaca
     8101 gctgaaaacg ttgcgaagag acttcaaaac cgggcttctg caagaggatc caagtgtaag
     8161 attatggcca tggatgattt ccctatcgag gagttaggca atgagaagaa tgtcgtagtc
     8221 ttggttagta cagctggtca aggtgagttt cctcaaaatg gacgtgagtt ttgggaagca
     8281 atcaaaggtg cagatcttaa cttgagcgaa ttgaagtatg gtgtatttgg ttttggcgat
     8341 tatgaatatt ggcctcgtaa ggaagataaa atttattata accgcccagg taagcaatta
     8401 gatgctcgct ttgttgaatt gggtgctgct ccattggtaa ccttaggact tggtaatgac
     8461 caggatccag atggatggga aacagcatat aacctttggg agcctgagct ttggaaagct
     8521 cttggattgg acaatgtcga aattgatatt gatgaaccta agcctatcac taatgaagac
     8581 attaaacaag cttcgaactt tttgcgtggt accatttttg aaggattggc cgatgaaagt
     8641 actggtgctt tggccgaaag tgactgtcaa cttaccaaat tccatggtat ttatatgcaa
     8701 gatgatcgtg acattcgtga tgagcgtaaa aaacaaggtt tggaacctgc ttatggtttc
     8761 atgattcgcg ccagaatgcc tgctggtgtc tgtacaccag aacaatggat tgctatggat
     8821 gatatttcta cgaaatgggg taatcacacc cttaagatta cgactcgtca aacgttccaa
     8881 tggcatggtg ttttgaagaa gaatttgcgt aataccattc gtaacatcgt taaagttttc
     8941 cttactactc ttggtgcttg tggtgatgtt gctcgtaatg tcacttgttc gtcgactccc
     9001 aacaatgaaa caattcacga tcaactgttt gctgtatcta agcaaatcag taatgaattg
     9061 cttcctacta catcctctta tcatgaaatt tggatcgagg accccgaaac tgtcgaaaag
     9121 agaaaggttg ccggtgaggc cgttcaggat gtcgagcctt tatatggtcc tacatatctg
     9181 cccagaaaat tcaaggttgg tgtggctgct cccccttaca atgatgtcga tgtttatacc
     9241 aatgatgttg ctttaatcgc catcattgaa aatgacaaag tcctaggttt caatgttggt
     9301 attggtggtg gtatgggcac aacccataac aacaagaaaa catatcctcg tttggctaca
     9361 gttgttggct atgttctaac tgataagatt atggaggtgg ttaaagctat tttgattgtc
     9421 cagcgtgata acggtgatcg tgaaaatcgt aagcacgctc gtcttaagta taccgttgat
     9481 actcttggtg tttctacctt cgtcgagaag gtcgaggagg ttttgggtta caaatttgaa
     9541 gaggcacgtg atcatcccca attcattaaa aaccacgatg acttcgaagg ctggcacaag
     9601 acagagaaaa acaaatactg gcgttctata tttgttgaaa acggtcgtat tgagaacaat
     9661 ggtattttac agtttaagac cggtttgcgt gaactcgcag aacgtctata tactgaaaaa
     9721 tctgaggctg aattccgttt gactgccaac cagcatgtca tcttatttaa tgttgctgaa
     9781 aatgagctcg gttggattaa tgagcatatg gccaaatata aacttgataa taatgcattc
     9841 tctggcttac gtctttcttc tgcagcctgc gttgctctcc ctacttgtgg tttagccatg
     9901 gctgaaagtg aacgttatct tcccaagttg attacgaaag tagaagaaat tgtttacgaa
     9961 gctggtcttc agaaagatag tattgttatg cgtatgactg gttgtccaaa tggatgctct
    10021 cgtccatggg ttgctgaaat tgcgtgtgtc ggtaaggcac caaatactta caatctaatg
    10081 cttggtggag gattttacgg tcagagatta aacaagctat atcgctcttc cgttcaagaa
    10141 aaagaaatcc tcaatttgtt gcgtccgctt atcaagcgtt acgccttgga gcgtgaagat
    10201 ggtgaacact ttggtgattg ggttattcgt gcaggtatca ttacagctgt tgaaaatgga
    10261 ggagctaatg gtgctgttca tgaaggggta tcacctgaag cgttt
//
