LOCUS       sequence-224157-        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-pomMet14
FEATURES             Location/Qualifiers
     source          1..6489
                     /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
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2019..2038
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     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..3371)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      3972..4849
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(4860..5373)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5055..5076)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5267..5286
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5395..5836
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    5395..5512
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     5807..5830
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     5868..5887
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5868..5886
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aaaggatttg attactttga agaagggcga 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 ggccaggaac 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 ctagaagaac
     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 tacgggaggg
     1501 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1561 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1621 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1681 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     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 aataccgcgc 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 tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat
     3361 gatgaattga aaagctagct tatcgatgat aagctgtcaa acatgagaat taattccacg
     3421 gactatagac tatacctagt atactccgtc tactgtacga tacacttccg ctcaggtcct
     3481 tgtcctttaa cgaggcctta ccactctttt gttactctat tgatccagct cagcaaaggc
     3541 agtgtgatct aagattctat cttcgcgatg tagtaaaact agctagaccg agaaagagac
     3601 tagaaatgca aaaggcactt ctacaatggc tgccatcatt attatccgat gtgacgctgc
     3661 agcttctcaa tgatattcga atacgctttg aggagataca gcctaatatc cgacaaactg
     3721 ttttacagat ttacgatcgt acttgttacc catcattgaa ttttgaacat ccgaacctgg
     3781 gagttttccc tgaaacagat agtatatttg aacctgtata ataatatata gtctagcgct
     3841 ttacggaaga caatgtatgt atttcggttc ctggagaaac tattgcatct attgcatagg
     3901 taatcttgca cgtcgcatcc ccggttcatt ttctgcgttt ccatcttgca cttcaatagc
     3961 atatctttgt taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag
     4021 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     4081 aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc
     4141 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     4201 cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc
     4261 atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc
     4321 gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa
     4381 ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt
     4441 actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat
     4501 tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct
     4561 tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg
     4621 aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt
     4681 ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg
     4741 caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata
     4801 tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaatg ggaagctcca
     4861 ccccggttga taatcagaaa agccccaaaa acaggaagat tgtataagca aatatttaaa
     4921 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     4981 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     5041 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     5101 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     5161 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     5221 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     5281 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     5341 ccgccgcgct taatgcgccg ctacagggcg cgtggggatc gatccactag tccacggatt
     5401 agaagccgcc gagcgggtga cagccctccg aaggaagact ctcctccgtg cgtcctcgtc
     5461 ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc cgcactgctc cgaacaataa
     5521 agattctaca atactagctt ttatggttat gaagaggaaa aattggcagt aacctggccc
     5581 cacaaacctt caaatgaacg aatcaaatta acaaccatag gatgataatg cgattagttt
     5641 tttagcctta tttctggggt aattaatcag cgaagcgatg atttttgatc tattaacaga
     5701 tatataaatg caaaaactgc ataaccactt taactaatac tttcaacatt ttcggtttgt
     5761 attacttctt attcaaatgt aataaaagta tcaacaaaaa attgttaata tacctctata
     5821 ctttaacgtc aaggagaaaa aaccccggat cggactacta gcagctgtaa tacgactcac
     5881 tatagggaat attaagctcg cccttatggc tactaacatt acatttcacc ctggttccgt
     5941 cactaaagaa gaacgtatca aatttgtggg tcatcctggt atgaccattt ggatgactgg
     6001 attatctgcg tctggaaagt ctacaatcgc ttgtgctttg gaacagtatt tacttcagcg
     6061 tggagttaca acttaccgtt tggatggtga taatgttcgt ttcggcttaa acagtgatct
     6121 tggatttagt gaacaagatc gtaacgaaaa cattcgtcgc ataggtcatg ttgccaagct
     6181 tttcgcagat gcctgtgtag ttgcagtaac ttccttcatt tctccttatc gcaaggatcg
     6241 tgatcaagca cgcgaatttc ataaaaaaga tggacttcct tttattgagg tttatgttga
     6301 atgtccagtg gaagttgccg agcagcgtga tccaaagggt ttgtacaagc gtgctcgtgc
     6361 tggtgagatt aaagaattta caggaatttc tgctccctat gaggctccca tttctcctga
     6421 aatcgtggtt tcttctcata ctcaatcgat tgaagagtgt gttgaaaaaa tcgtcaatta
     6481 cttgttgga
//
