LOCUS       sequence-202685-        6492 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 6492)
  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 6492)
  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-pomMet14
FEATURES             Location/Qualifiers
     source          1..6492
                     /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 cactatatgg ctactaacat tacatttcac cctggttccg tcactaaaga agaacgtatc
     5941 aaatttgtgg gtcatcctgg tatgaccatt tggatgactg gattatctgc gtctggaaag
     6001 tctacaatcg cttgtgcttt ggaacagtat ttacttcagc gtggagttac aacttaccgt
     6061 ttggatggtg ataatgttcg tttcggctta aacagtgatc ttggatttag tgaacaagat
     6121 cgtaacgaaa acattcgtcg cataggtcat gttgccaagc ttttcgcaga tgcctgtgta
     6181 gttgcagtaa cttccttcat ttctccttat cgcaaggatc gtgatcaagc acgcgaattt
     6241 cataaaaaag atggacttcc ttttattgag gtttatgttg aatgtccagt ggaagttgcc
     6301 gagcagcgtg atccaaaggg tttgtacaag cgtgctcgtg ctggtgagat taaagaattt
     6361 acaggaattt ctgctcccta tgaggctccc atttctcctg aaatcgtggt ttcttctcat
     6421 actcaatcga ttgaagagtg tgttgaaaaa atcgtcaatt acttgttgga aaaggatttg
     6481 attactttga ag
//
