LOCUS       sequence-208919-       10209 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 10209)
  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 10209)
  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-cerMET5
FEATURES             Location/Qualifiers
     source          1..10209
                     /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             4868..4909
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4919..4936
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4957..5204
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4973..4991)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5281..5298)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5452..6040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5532..5551)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6211..7002,7003..7071))
                     /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     6834..6853
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(7167..7970)
                     /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(7971..8186)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      8787..9664
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(9675..10188)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9870..9891)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10082..10101
                     /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 tatgactgct tctgacctct tgacgctccc
      541 acaattgttg gcgcaatatt cctctagtgc tccccaaaat aaagtgttct acactactag
      601 cacaaaaaat agtcattcat ccttcaaagg gctagaatct gttgccacag atgctactca
      661 tctattgaac aatcaagatc cattgaatac catcaaagat caactctcta aggacatttt
      721 aactaccgtt tttacagatg aaactacttt gatcaaatcc atccaccatc tatattctct
      781 tcctaataaa cttccattgg tgattacagt ggatttgaat ttgcaagatt attccgcaat
      841 tcctgcgtta aaggatcttt ctttccccat cttaatatct tctgatttgc aaactgcaat
      901 ttcaaacgcg gactcttctt ataagattgc tacaagttct ctgaccccag ttttccattt
      961 tcttaacttg gagaaaattg gcacgagtac agctattgaa caagacatcg actttcctac
     1021 cttggagatt gctaacgaag agactaaagt agcattatca gaagctactg attcattgac
     1081 aaattttgaa ctggttaaag gtaaagagtc aattaccact gtcattgtta acctgtcacc
     1141 atatgacgcg gaattcagta gcgtattgcc ttcaaacgta ggattgatca agattagagt
     1201 atacagacca tggaattttt ccaagttctt ggaaatactg ccatcttctg ttaccaaaat
     1261 cgccgtttta cagggtgtct ctaagaaatc gcaatcaaac gaatttcaac catttctttt
     1321 ggacttcttt ggcaatttta atgaattagt ttctaggaac atcgagcagg tagtgttgac
     1381 taatattggt aatgtgaatg attatggcaa cgttatcaac accgtcatat cgaatattaa
     1441 caagaaggaa ccagataata acttattttt aggtgaatcc aatgaaaagg ctgaggaaca
     1501 agctgaagtt acccaactta tttcttctgt cgaaaaagtt gtgaacttag aagacgccta
     1561 tatcaaagtg ctaaaacagt tattttcatc aaatctacaa attttgaatc aattttccag
     1621 tgagacaatt gaaccaagta atccagaatt tggttttgga cgctttttaa aacaagaagc
     1681 ccagcgtgaa gaattgatca gcttagcaaa aacctctctt gatccaagtc tttacttgtc
     1741 cgaggatgca aataaaattg ttcaactatt atctaaatgg ttgtcattca atggacgcga
     1801 tcttgacgaa gctcaattac aagaggccaa tgcgacaggt ttggaaatat ttcagttatt
     1861 acaatctaat caagattcct ctactgtctt aaaattctta aagatagctc caacaagcga
     1921 ttcttttatt ttcaaatcaa gctggctaat tggctccgat gcctggtctt atgatttggg
     1981 tcactcaggt attcaacagg ttttatcctc ccgtaaaaac attaatgttt tattgattga
     2041 ttcagagcca tatgaccata gaaagcaaaa ccaggataga aagaaagatg ttggtttgta
     2101 cgccatgaat tattacagtg cctatgttgc ctctgtagca gtatatgctt cttacaccca
     2161 actattgact gcaataatag aggcatctaa atacaatggt ccttctattg tcttggctta
     2221 tttgccgtat aattccgaaa atgatactcc attagaagtc ttaaaggaaa ccaaaaacgc
     2281 cgttgaatct ggttactggc tattgtatag gtttaatcct gtttgtgacg atccatcaac
     2341 agataaggag gcatttagct tggattcttc ggttatcaga aaacaattac aggacttctt
     2401 agaccgtgag aataagctta ccctattaac cagaaaagat ccatctttgt caagaaatct
     2461 aaagcaatct gctggtgatg cgttgacaag gaaacaagaa aaaagaagca aggctgcctt
     2521 cgatcagtta ttggagggtt tgtccggccc accgctacac gtctattatg cttctgacgg
     2581 tggtaatgct gcaaacttgg caaagagact agcagcaagg gcatctgcaa gaggtttaaa
     2641 ggctactgtt ctgtccatgg atgacattat tttggaagaa ttacctggtg aagagaatgt
     2701 agttttcatt acttccacgg ctgggcaagg tggattcccc caagacggta agtctttctg
     2761 ggaagctctg aaaaatgaca ccgacttgga tttagctagt ttgaatgttg ctgtttttgg
     2821 tctcggtgat tctgagtatt ggccacgtaa agaagataaa cattatttta acaagccttc
     2881 acaggattta tttaagcgct tggaattatt gagtgccaaa gccctaattc ccttgggact
     2941 gggtgatgat caagatgctg atggtttcca aactgcttat tctgagtggg aacctaaatt
     3001 atgggaagct cttggtgttt ccggcgctgc tgttgatgat gagccaaaac ctgttacaaa
     3061 cgaggatatt aagagagaat ctaatttctt gagaggtact atcagtgaga acttaaagga
     3121 tacttcttca ggtggtgtta ctcacgctaa tgaacaatta atgaaatttc acggtattta
     3181 cacccaagac gatcgtgaca ttagagaaat acgtaagtca caaggcttag agccatacta
     3241 tatgtttatg gcaagagctc gtttaccagg tggtaagacc actccacaac aatggcttgc
     3301 tctgggtcac ttatctgata cttcaggcaa tggtaccctg aaattaacaa caagggcaac
     3361 cttccagatt catggtgtgc taaagaagaa cttgaaacac acattgagag gaatgaatgc
     3421 agttcttatg gatacattag ctgctgcagg tgacgtgaac agaaatgtca tggtttctgc
     3481 tctaccaacc aatgccaagg ttcaccaaca aatcgctgat atgggaaaat tgatttctga
     3541 tcatttctta ccaaagacta cggcctacca cgaagtttgg ctggagggcc cagaagaaca
     3601 ggacgatgat ccatcctggc catctatttt tgaaaacaga aaagatggtc caagaaaaaa
     3661 gaagactcta gttagcggta acgctttggt cgatattgaa ccaatttacg gtccaactta
     3721 tctgccaaga aagtttaaat tcaacatcgc cgttcctcca tataacgatg tggatgtatt
     3781 atctatcgat gtcggtttag ttgctatagt taacccagaa actcaaatcg tggagggtta
     3841 taatgttttt gttggtggtg gtatgggtac cactcataac aacaagaaaa cttacccaag
     3901 attagggtca tgcttaggtt ttgttaaaac tgaagacatt attccaccac ttgaaggtat
     3961 cgttattgtc caaagagatc acggtgaccg taaagaccgt aagcatgctc gtttaaagta
     4021 tactgtagac gatatgggtg tcgaaggctt caagcaaaaa gtggaggaat actggggtaa
     4081 gaaattcgag cctgagagac catttgagtt taaatctaat attgattact ttggatggat
     4141 taaagatgaa actgggttaa gccactttac cgcatttatt gaaaatggta gggttgaaga
     4201 tacaccagat ttgcctcaaa agacaggtat tagaaaggtt gctgaataca tgcttaagac
     4261 taattctggt catttcagat tgactggtaa tcaacatttg gttatctcta atattacaga
     4321 tgaacatgtt gctggaataa aatctatttt aaagacctat aaattggata acaccgattt
     4381 cagcggtttg agattatctt catcttcctg tgttggtttg ccaacatgtg gtttagcgtt
     4441 tgccgaatct gaacgtttcc tacctgacat tattactcag ttggaagatt gtttagaaga
     4501 gtatggttta cgccgtgatt ccattattat gagaatgact ggttgcccta acggttgttc
     4561 tcgtccatgg ctaggtgagt tagctcttgt tggtaaagct ccacatactt ataacttgat
     4621 gcttggtggt ggttacctcg gccaaaggct aaacaaattg tataaggcca atgtgaagga
     4681 tgaggaaatt gtcgactaca tcaaaccatt gtttaaaagg tatgctttag aaagagaaga
     4741 aggggaacac tttggtgatt tctgtataag agtaggtatc attaaaccaa ccaccgaggg
     4801 taaatacttc catgaagatg tgtctgaaga tgcctataag ggcgagcttc gaggtcaccc
     4861 attcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca
     4921 tcatcaccat caccattgag tttctagagg gccgcatcat gtaattagtt atgtcacgct
     4981 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
     5041 aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat
     5101 ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa
     5161 ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcaagctg cggccctgca
     5221 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5281 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5341 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5401 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5461 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5521 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5581 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5641 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5701 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5761 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5821 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5881 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5941 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6001 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6061 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6121 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6181 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6241 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6301 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6361 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6421 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6481 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6541 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6601 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6661 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6721 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6781 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6841 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6901 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6961 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7021 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7081 ttttcaatgg gtaataactg atataattaa attgaagctc taatttgtga gtttagtata
     7141 catgcattta cttataatac agttttttag ttttgctggc cgcatcttct caaatatgct
     7201 tcccagcctg cttttctgta acgttcaccc tctaccttag catcccttcc ctttgcaaat
     7261 agtcctcttc caacaataat aatgtcagat cctgtagaga ccacatcatc cacggttcta
     7321 tactgttgac ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt cataatcaac
     7381 caatcgtaac cttcatctct tccacccatg tctctttgag caataaagcc gataacaaaa
     7441 tctttgtcgc tcttcgcaat gtcaacagta cccttagtat attctccagt agatagggag
     7501 cccttgcatg acaattctgc taacatcaaa aggcctctag gttcctttgt tacttcttct
     7561 gccgcctgct tcaaaccgct aacaatacct gggcccacca caccgtgtgc attcgtaatg
     7621 tctgcccatt ctgctattct gtatacaccc gcagagtact gcaatttgac tgtattacca
     7681 atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt acttggcgga taatgccttt
     7741 agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga tatccacatg tgtttttagt
     7801 aaacaaattt tgggacctaa tgcttcaact aactccagta attccttggt ggtacgaaca
     7861 tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga tattaaatag cttggcagca
     7921 acaggactag gatgagtagc agcacgttcc ttatatgtag ctttcgacat gatttatctt
     7981 cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga atactgggca atttcatgtt
     8041 tcttcaacac tacatatgcg tatatatacc aatctaagtc tgtgctcctt ccttcgttct
     8101 tccttctgtt cggagattac cgaatcaaaa aaatttcaag gaaaccgaaa tcaaaaaaaa
     8161 gaataaaaaa aaaatgatga attgaaaagc tagcttatcg atgataagct gtcaaacatg
     8221 agaattaatt ccacggacta tagactatac ctagtatact ccgtctactg tacgatacac
     8281 ttccgctcag gtccttgtcc tttaacgagg ccttaccact cttttgttac tctattgatc
     8341 cagctcagca aaggcagtgt gatctaagat tctatcttcg cgatgtagta aaactagcta
     8401 gaccgagaaa gagactagaa atgcaaaagg cacttctaca atggctgcca tcattattat
     8461 ccgatgtgac gctgcagctt ctcaatgata ttcgaatacg ctttgaggag atacagccta
     8521 atatccgaca aactgtttta cagatttacg atcgtacttg ttacccatca ttgaattttg
     8581 aacatccgaa cctgggagtt ttccctgaaa cagatagtat atttgaacct gtataataat
     8641 atatagtcta gcgctttacg gaagacaatg tatgtatttc ggttcctgga gaaactattg
     8701 catctattgc ataggtaatc ttgcacgtcg catccccggt tcattttctg cgtttccatc
     8761 ttgcacttca atagcatatc tttgttaacg aagcatctgt gcttcatttt gtagaacaaa
     8821 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     8881 agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa
     8941 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     9001 agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt
     9061 gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt
     9121 ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg
     9181 ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact
     9241 ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg
     9301 catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat
     9361 agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta
     9421 tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag
     9481 ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga
     9541 ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata
     9601 tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg
     9661 caatgggaag ctccaccccg gttgataatc agaaaagccc caaaaacagg aagattgtat
     9721 aagcaaatat ttaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     9781 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     9841 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     9901 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     9961 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
    10021 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
    10081 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
    10141 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtgg ggatcgatcc
    10201 actagtcca
//
