LOCUS       sequence-208887-        6681 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 6681)
  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 6681)
  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-pomMet16
FEATURES             Location/Qualifiers
     source          1..6681
                     /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             1340..1381
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1391..1408
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1429..1676
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1445..1463)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1753..1770)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1924..2512)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2004..2023)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2683..3474,3475..3543))
                     /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     3306..3325
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(3639..4442)
                     /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(4443..4658)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5259..6136
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6147..6660)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6342..6363)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6554..6573
                     /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 tatgagctca atcgatacgc ctgctaataa
      541 attgcaagcg aaaaccttgt ttcacccaga acatctcgaa tacatcaaca agcagttatc
      601 ggagcttagt cctcaagata tcttaaaatg gtgccgttgg acacttcctt ctttgtttca
      661 aacatctgcg ttaggactgt ccgggcttgt tattatggac atgttatcga aaatggatat
      721 gaatgttcct ctgattttta tcaacacact tcatcatttc ccagaaacac tggatcttct
      781 tgaaaaagtc aaaactaagt atcctaacgt gcctgttcac gtctatcgtt gtgctgaagc
      841 agccaacgaa aaggaatttg ctcagaagtt tggtgagaag ctttgggaga cagatgaatc
      901 tcggtatgac tttttagtca aagtggaacc agcaagtcgt gcttattctg acttaaatgt
      961 acttgctgtg tttactggcc gccgccgtag tcaaggtgga gagcgcggat ctttgcccat
     1021 tgttcagctg gatggaccag tattaaagat caaccccttg gctaactggt cttttactga
     1081 agttcataat tatattatta caaacaacgt tccatataat gaacttctga acaagggtta
     1141 cagatcagtt ggtgactggc actctactca accagttaga gaaggtgaag atgaacgtgc
     1201 cggacgttgg cgcggtcgag agaaaacaga atgtggatta cacagtcatc ctcaaagtaa
     1261 attcgcacaa tatatggcgg agcttaaaaa gaaggagacg gctgatcaga agggcgagct
     1321 tcgaggtcac ccattcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac
     1381 gcgtaccggt catcatcacc atcaccattg agtttctaga gggccgcatc atgtaattag
     1441 ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag
     1501 ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa
     1561 cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca
     1621 ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcaagc
     1681 tgcggccctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1741 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1801 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1861 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1921 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1981 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2041 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2101 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2161 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2221 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2281 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2341 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2401 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2461 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2521 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2581 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2641 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2701 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2761 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2821 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2881 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2941 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3001 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3061 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3121 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3181 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3241 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3301 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3361 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3421 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3481 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3541 catactcttc ctttttcaat gggtaataac tgatataatt aaattgaagc tctaatttgt
     3601 gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt
     3661 ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt
     3721 ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca
     3781 tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt
     3841 gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag
     3901 ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca
     3961 gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt
     4021 gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt
     4081 gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg
     4141 actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg
     4201 gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa gatatccaca
     4261 tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag taattccttg
     4321 gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat
     4381 agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac
     4441 atgatttatc ttcgtttcct gcaggttttt gttctgtgca gttgggttaa gaatactggg
     4501 caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc
     4561 ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca aggaaaccga
     4621 aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa gctagcttat cgatgataag
     4681 ctgtcaaaca tgagaattaa ttccacggac tatagactat acctagtata ctccgtctac
     4741 tgtacgatac acttccgctc aggtccttgt cctttaacga ggccttacca ctcttttgtt
     4801 actctattga tccagctcag caaaggcagt gtgatctaag attctatctt cgcgatgtag
     4861 taaaactagc tagaccgaga aagagactag aaatgcaaaa ggcacttcta caatggctgc
     4921 catcattatt atccgatgtg acgctgcagc ttctcaatga tattcgaata cgctttgagg
     4981 agatacagcc taatatccga caaactgttt tacagattta cgatcgtact tgttacccat
     5041 cattgaattt tgaacatccg aacctgggag ttttccctga aacagatagt atatttgaac
     5101 ctgtataata atatatagtc tagcgcttta cggaagacaa tgtatgtatt tcggttcctg
     5161 gagaaactat tgcatctatt gcataggtaa tcttgcacgt cgcatccccg gttcattttc
     5221 tgcgtttcca tcttgcactt caatagcata tctttgttaa cgaagcatct gtgcttcatt
     5281 ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     5341 tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt
     5401 catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
     5461 ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta
     5521 tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc
     5581 ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc
     5641 actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa
     5701 aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt
     5761 tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga
     5821 acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct
     5881 attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca
     5941 ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat
     6001 aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt
     6061 tatataggga tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg
     6121 aagcggtatt cgcaatggga agctccaccc cggttgataa tcagaaaagc cccaaaaaca
     6181 ggaagattgt ataagcaaat atttaaattg taagcgttaa tattttgtta aaattcgcgt
     6241 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     6301 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     6361 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     6421 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     6481 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     6541 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     6601 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     6661 ggggatcgat ccactagtcc a
//
