LOCUS       sequence-208850-        6675 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 6675)
  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 6675)
  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-pomMet8
FEATURES             Location/Qualifiers
     source          1..6675
                     /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             1334..1375
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1385..1402
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1423..1670
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1439..1457)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1747..1764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1918..2506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1998..2017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2677..3468,3469..3537))
                     /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     3300..3319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(3633..4436)
                     /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(4437..4652)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5253..6130
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6141..6654)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6336..6357)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6548..6567
                     /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 tatggcttca aaatatcagt ctgtgcagcc
      541 tggtggatca ttaattattg cttggcgtgc acaaggtaaa aaagtattaa ttgttggagg
      601 tggcgttgtg gcagctgggc gtatattgca tgttttgaat gcggatgccc atgttatcgt
      661 tgtaagtcca aaagctggtc tttgtaaaga agtggcttgg agaatacagg agaagcaagt
      721 tgaatggcga gacaggggat ttctagtcga agatttgtct gacgatgtaa acatggtact
      781 tacagccata gatgatccca gcttgtcttc cgaaatttat aagttatgca aaagcaaaaa
      841 aatcccggtc aatgcggcgg atattccacc tgaatgtgat ttctactttg ggtctgaaat
      901 ccgtaacggc cctctacaaa ttatggttag caccaatggt aaaggtccaa aacttgccag
      961 tttaatacga aaaaaaatcg aaagttcaat caatccagct actggtatgg cattggaaaa
     1021 aactggcctt ttacgacaaa aattacgaga cattgttccc gaacctgaaa acagtcgcaa
     1081 aagaatgaga tggatgattg agatttgcga actgtggtct ttggaggaac ttgccatgct
     1141 cgacgaaaat ctcatcaacc gacttttggg atacttccca aaaaaaacac ctagttatcg
     1201 agaaataacg actccagcct acagcaagat agacaactac atatggtttg gagcaattat
     1261 aatttctggg gctgttctcc tcaaacatgt ttcaaaagct cgtaagggcg agcttcgagg
     1321 tcacccattc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     1381 cggtcatcat caccatcacc attgagtttc tagagggccg catcatgtaa ttagttatgt
     1441 cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac
     1501 aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat
     1561 ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac
     1621 tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc aagctgcggc
     1681 cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1741 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1801 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1861 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1921 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1981 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2041 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2101 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2161 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2221 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2281 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2341 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2401 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2461 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2521 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2581 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2641 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2701 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2761 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2821 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2881 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2941 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3001 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3061 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3121 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3181 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3241 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3301 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3361 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3421 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3481 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3541 cttccttttt caatgggtaa taactgatat aattaaattg aagctctaat ttgtgagttt
     3601 agtatacatg catttactta taatacagtt ttttagtttt gctggccgca tcttctcaaa
     3661 tatgcttccc agcctgcttt tctgtaacgt tcaccctcta ccttagcatc ccttcccttt
     3721 gcaaatagtc ctcttccaac aataataatg tcagatcctg tagagaccac atcatccacg
     3781 gttctatact gttgacccaa tgcgtctccc ttgtcatcta aacccacacc gggtgtcata
     3841 atcaaccaat cgtaaccttc atctcttcca cccatgtctc tttgagcaat aaagccgata
     3901 acaaaatctt tgtcgctctt cgcaatgtca acagtaccct tagtatattc tccagtagat
     3961 agggagccct tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc ctttgttact
     4021 tcttctgccg cctgcttcaa accgctaaca atacctgggc ccaccacacc gtgtgcattc
     4081 gtaatgtctg cccattctgc tattctgtat acacccgcag agtactgcaa tttgactgta
     4141 ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa aattgtactt ggcggataat
     4201 gcctttagcg gcttaactgt gccctccatg gaaaaatcag tcaagatatc cacatgtgtt
     4261 tttagtaaac aaattttggg acctaatgct tcaactaact ccagtaattc cttggtggta
     4321 cgaacatcca atgaagcaca caagtttgtt tgcttttcgt gcatgatatt aaatagcttg
     4381 gcagcaacag gactaggatg agtagcagca cgttccttat atgtagcttt cgacatgatt
     4441 tatcttcgtt tcctgcaggt ttttgttctg tgcagttggg ttaagaatac tgggcaattt
     4501 catgtttctt caacactaca tatgcgtata tataccaatc taagtctgtg ctccttcctt
     4561 cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat ttcaaggaaa ccgaaatcaa
     4621 aaaaaagaat aaaaaaaaaa tgatgaattg aaaagctagc ttatcgatga taagctgtca
     4681 aacatgagaa ttaattccac ggactataga ctatacctag tatactccgt ctactgtacg
     4741 atacacttcc gctcaggtcc ttgtccttta acgaggcctt accactcttt tgttactcta
     4801 ttgatccagc tcagcaaagg cagtgtgatc taagattcta tcttcgcgat gtagtaaaac
     4861 tagctagacc gagaaagaga ctagaaatgc aaaaggcact tctacaatgg ctgccatcat
     4921 tattatccga tgtgacgctg cagcttctca atgatattcg aatacgcttt gaggagatac
     4981 agcctaatat ccgacaaact gttttacaga tttacgatcg tacttgttac ccatcattga
     5041 attttgaaca tccgaacctg ggagttttcc ctgaaacaga tagtatattt gaacctgtat
     5101 aataatatat agtctagcgc tttacggaag acaatgtatg tatttcggtt cctggagaaa
     5161 ctattgcatc tattgcatag gtaatcttgc acgtcgcatc cccggttcat tttctgcgtt
     5221 tccatcttgc acttcaatag catatctttg ttaacgaagc atctgtgctt cattttgtag
     5281 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     5341 cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt
     5401 tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     5461 ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc
     5521 ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat
     5581 tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta
     5641 ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc
     5701 ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga
     5761 taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa
     5821 agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg
     5881 tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat
     5941 gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa
     6001 tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata
     6061 gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg
     6121 tattcgcaat gggaagctcc accccggttg ataatcagaa aagccccaaa aacaggaaga
     6181 ttgtataagc aaatatttaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     6241 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     6301 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     6361 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     6421 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     6481 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     6541 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     6601 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtggggat
     6661 cgatccacta gtcca
//
