LOCUS       sequence-321477-        5182 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Yeast display plasmid for expression of C-terminal fusions. 
            Modification of pJYDC1 vector with ALFA tag in MCS..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5182)
  AUTHORS   Zahradnik J, Dey D, Marciano S, Schreiber G
  TITLE     An enhanced yeast display platform demonstrates the binding 
            plasticity under various selection pressures
  JOURNAL   bioRxiv Preprint 2020
REFERENCE   2  (bases 1 to 5182)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            Preprint 2020"
COMMENT     Sequence Label: pJYDC2
FEATURES             Location/Qualifiers
     source          1..5182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(121..795)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(796..1076)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(1097..1116)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1177..1195)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    1236..1740
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        1777..1881
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1882..2697
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1882..2697
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1955..1974)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2868..3456
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3357..3376
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3639..4080
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    3639..3756
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     4051..4074
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             4850..4861
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             4862..4888
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5022..5060
                     /codon_start=1
                     /product="rationally designed epitope tag that forms a 
                     stable alpha-helix (Gotzke et al., 2019)"
                     /label=ALFA-tag
                     /note="It may be beneficial to flank the ALFA-tag sequence 
                     with proline residues to preserve the helical structure."
                     /translation="SRLEEELRRRLTE"
ORIGIN
        1 taatatcctt ttctattttt cgttccgtta ccaactttac acatactttt gcggtatttc
       61 acaccgcata gatcggcaag tgcacaaaca atacttaaat aaatactact cagtaataac
      121 ctatttctta gcatttttga cgaaatttgc tattttgtta gagtctttta caccatttgt
      181 ctccacacct ccgcttacat caacaccaat aacgccattt aatctaagcg catcaccaac
      241 attttctggc gtcagtccac cagctaacat aaaatgtaag ctttcggggc tctcttgcct
      301 tccaacccag tcagaaatcg agttccaatc caaaagttca cctgtcccac ctgcttctga
      361 atcaaacaag ggaataaacg aatgaggttt ctgtgaagct gcactgagta gtatgttgca
      421 gtcttttgga aatacgagtc ttttaataac tggcaaaccg aggaactctt ggtattcttg
      481 ccacgactca tctccatgca gttggacgat atcaatgccg taatcattga ccagagccaa
      541 aacatcctcc ttaggttgat tacgaaacac gccaaccaag tatttcggag tgcctgaact
      601 atttttatat gcttttacaa gacttgaaat tttccttgca ataaccgggt caattgttct
      661 ctttctattg ggcacacata taatacccag caagtcagca tcggaatcta gagcacattc
      721 tgcggcctct gtgctctgca agccgcaaac tttcaccaat ggaccagaac tacctgtgaa
      781 attaataaca gacatactcc aagctgcctt tgtgtgctta atcacgtata ctcacgtgct
      841 caatagtcac caatgccctc cctcttggcc ctctcctttt cttttttcga ccgaattaat
      901 tcttaatcgg caaaaaaaga aaagctccgg atcaagattg tacgtaaggt gacaagctat
      961 ttttcaataa agaatatctt ccactactgc catctggcgt cataactgca aagtacacat
     1021 atattacgat gctgttctat taaatgcttc ctatattata tatatagtaa tgtcgtgatc
     1081 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     1141 cgccaacacc cgctgacgcg ccctcgagac gaaagggcct cgtgatacgc ctatttttat
     1201 aggttaatgt catgataata atggtttctt agacggatcg cttgcctgta acttacacgc
     1261 gcctcgtatc ttttaatgat ggaataattt gggaatttac tctgtgttta tttattttta
     1321 tgttttgtat ttggatttta gaaagtaaat aaagaaggta gaagagttac ggaatgaaga
     1381 aaaaaaaaat aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt
     1441 tattagacaa gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta
     1501 aaatcacagg attttcgtgt gtggtcttct acacagacaa gatgaaacaa ttcggcatta
     1561 atacctgaga gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt
     1621 cttttacatc ttcggaaaac aaaaactatt ttttctttaa tttctttttt tactttctat
     1681 ttttaattta tatatttata ttaaaaaatt taaattataa ttatttttat agcacgtgat
     1741 gaaaaggacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1801 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1861 taatattgaa aaaggaagag tatgagccat attcaacggg aaacgtcttg ctcgaggccg
     1921 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     1981 gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt
     2041 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     2101 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     2161 gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat
     2221 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     2281 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     2341 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     2401 cctgttgaac aagtctggaa agaaatgcat aagcttttgc cattctcacc ggattcagtc
     2461 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     2521 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     2581 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     2641 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaactg
     2701 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     2761 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     2821 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2881 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2941 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3001 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3061 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3121 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3181 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3241 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3301 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3361 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3421 ttgtgatgct cgtcaggggg gcggagccta tgaaaaaacg ccagcaacgc ggcactttgt
     3481 aactgagctg tcattttgct ggcccagctg gtcgaccgcg gctaagcagt actggtaccg
     3541 ttgtggaaat gtaaagagcc ccattatctt agcctaaaaa aaccttctct ttggaacttt
     3601 cagtaatacg cttaactgct cattgctata ttgaagtacg gattagaagc cgccgagcgg
     3661 gtgacagccc tccgaaggaa gactctcctc cgtgcgtcct cgtcttcacc ggtcgcgttc
     3721 ctgaaacgca gatgtgcctc gcgccgcact gctccgaaca ataaagattc tacaatacta
     3781 gcttttatgg ttatgaagag gaaaaattgg cagtaacctg gccccacaaa ccttcaaatg
     3841 aacgaatcaa attaacaacc ataggatgat aatgcgatta gttttttagc cttatttctg
     3901 gggtaattaa tcagcgaagc gatgattttt gatctattaa cagatatata aatgcaaaaa
     3961 ctgcataacc actttaacta atactttcaa cattttcggt ttgtattact tcttattcaa
     4021 atgtaataaa agtatcaaca aaaaattgtt aatatacctc tatactttaa cgtcaaggag
     4081 aaaaaacccc ggatcgaatt ccctacttca tacattttca attaagatga ggttcccatc
     4141 tattttcacc gctgttgttt ttgctgcttc ttctgctttg gctgctccag ctaatggtat
     4201 gttagaaaaa tttgttggca cctggaagat cgaatcctct gaaaattttg gtgaatactt
     4261 gaaggctatc ggtgccccaa aagaattggc tgatgctggt gatgctacta ctccagtctt
     4321 gtacatttct caaaaggatg gtgataagat gaccgtcaag attgaaaacg gtccaccaac
     4381 ttttttggat acccaagttt ctttcaagtt gggtgaagaa ttcgacgaat ttccatccga
     4441 tagaagaaag ggtgttaagt ccgttgttaa cttgtctggt gagaagttgg tttacgttca
     4501 aaagtgggat ggtaaagaaa ccacttacgt cagagaaatc aaggacggta aattggttgt
     4561 tactttgacc atgggtgatg ttgttgctgt tagatcttat agaagggcct ctgaagtttc
     4621 tgcacaggaa ctgacaacta tatgcgagca aatcccctca ccaactttag aatcgacgcc
     4681 gtactctttg tcaacgacta ctattttggc caacgggaag gcaatgcaag gagtttttga
     4741 atattacaaa tcagtaacgt ttgtcagtaa ttgcggttct cacccctcaa caactagcaa
     4801 aggcagcccc ataaacacac agtatgtttt taaggacaat agctcgacga ttgaaggtag
     4861 atacccatac gacgttccag actacgctct gcaggctagt ggtggaggag gctctggtgg
     4921 aggcggtagc ggaggcggag ggtcggctag ccatatgcac gatgaactat aatgaagcat
     4981 gcttaattaa ggcgccttat ctcgaggggg gcggatcccc atccagattg gaagaagaat
     5041 tgagaagaag attaaccgaa taatagagat ctgataacaa cagtgtagat gtaacaaaat
     5101 cgactttgtt cccactgtac ttttagctcg tacaaaatac aatatacttt tcatttctcc
     5161 gtaaacaaca tgttttccca tg
//
