LOCUS       sequence-314852-        5181 bp    DNA     circular SYN 15-DEC-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 5181)
  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 5181)
  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: "bioRxiv 
            Preprint 2020"
COMMENT     Sequence Label: pJYDC2
FEATURES             Location/Qualifiers
     source          1..5181
                     /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..1739
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        1776..1880
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1881..2696
                     /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             1881..2696
                     /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(1954..1973)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2867..3455
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3356..3375
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3638..4079
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    3638..3755
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     4050..4073
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             4849..4860
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             4861..4887
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5021..5059
                     /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 cccttgagac 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 aaaaaaaata aacaaaggtt taaaaaattt caacaaaaag cgtactttac atatatattt
     1441 attagacaag aaaagcagat taaatagata tacattcgat taacgataag taaaatgtaa
     1501 aatcacagga ttttcgtgtg tggtcttcta cacagacaag atgaaacaat tcggcattaa
     1561 tacctgagag caggaagagc aagataaaag gtagtatttg ttggcgatcc ccctagagtc
     1621 ttttacatct tcggaaaaca aaaactattt tttctttaat ttcttttttt actttctatt
     1681 tttaatttat atatttatat taaaaaattt aaattataat tatttttata gcacgtgatg
     1741 aaaaggaccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1801 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1861 aatattgaaa aaggaagagt atgagccata ttcaacggga aacgtcttgc tcgaggccgc
     1921 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     1981 ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc
     2041 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     2101 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     2161 catggttact caccactgcg atccccggga aaacagcatt ccaggtatta gaagaatatc
     2221 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     2281 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     2341 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     2401 ctgttgaaca agtctggaaa gaaatgcata agcttttgcc attctcaccg gattcagtcg
     2461 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     2521 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     2581 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     2641 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaactgt
     2701 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     2761 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2821 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2881 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2941 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3001 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3061 caccacctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3121 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3181 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3241 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3301 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3361 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3421 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcactttgta
     3481 actgagctgt cattttgctg gcccagctgg tcgaccgcgg ctaagcagta ctggtaccgt
     3541 tgtggaaatg taaagagccc cattatctta gcctaaaaaa accttctctt tggaactttc
     3601 agtaatacgc ttaactgctc attgctatat tgaagtacgg attagaagcc gccgagcggg
     3661 tgacagccct ccgaaggaag actctcctcc gtgcgtcctc gtcttcaccg gtcgcgttcc
     3721 tgaaacgcag atgtgcctcg cgccgcactg ctccgaacaa taaagattct acaatactag
     3781 cttttatggt tatgaagagg aaaaattggc agtaacctgg ccccacaaac cttcaaatga
     3841 acgaatcaaa ttaacaacca taggatgata atgcgattag ttttttagcc ttatttctgg
     3901 ggtaattaat cagcgaagcg atgatttttg atctattaac agatatataa atgcaaaaac
     3961 tgcataacca ctttaactaa tactttcaac attttcggtt tgtattactt cttattcaaa
     4021 tgtaataaaa gtatcaacaa aaaattgtta atatacctct atactttaac gtcaaggaga
     4081 aaaaaccccg gatcgaattc cctacttcat acattttcaa ttaagatgag gttcccatct
     4141 attttcaccg ctgttgtttt tgctgcttct tctgctttgg ctgctccagc taatggtatg
     4201 ttagaaaaat ttgttggcac ctggaagatc gaatcctctg aaaattttgg tgaatacttg
     4261 aaggctatcg gtgccccaaa agaattggct gatgctggtg atgctactac tccagtcttg
     4321 tacatttctc aaaaggatgg tgataagatg accgtcaaga ttgaaaacgg tccaccaact
     4381 tttttggata cccaagtttc tttcaagttg ggtgaagaat tcgacgaatt tccatccgat
     4441 agaagaaagg gtgttaagtc cgttgttaac ttgtctggtg agaagttggt ttacgttcaa
     4501 aagtgggatg gtaaagaaac cacttacgtc agagaaatca aggacggtaa attggttgtt
     4561 actttgacca tgggtgatgt tgttgctgtt agatcttata gaagggcctc tgaagtttct
     4621 gcacaggaac tgacaactat atgcgagcaa atcccctcac caactttaga atcgacgccg
     4681 tactctttgt caacgactac tattttggcc aacgggaagg caatgcaagg agtttttgaa
     4741 tattacaaat cagtaacgtt tgtcagtaat tgcggttctc acccctcaac aactagcaaa
     4801 ggcagcccca taaacacaca gtatgttttt aaggacaata gctcgacgat tgaaggtaga
     4861 tacccatacg acgttccaga ctacgctctg caggctagtg gtggaggagg ctctggtgga
     4921 ggcggtagcg gaggcggagg gtcggctagc catatgcacg atgaactata atgaagcatg
     4981 cttaattaag gcgccttatc tcgagggggg cggatcccca tccagattgg aagaagaatt
     5041 gagaagaaga ttaaccgaat aatagagatc tgataacaac agtgtagatg taacaaaatc
     5101 gactttgttc ccactgtact tttagctcgt acaaaataca atatactttt catttctccg
     5161 taaacaacat gttttcccat g
//
