LOCUS       sequence-314843-        5254 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast display platform plasmid N3 which allows for simultaneous 
            expression of the protein of interest at the N-terminus of Aga2p 
            yeast agglutinin and eUnaG2 or nbALFA detection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5254)
  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 5254)
  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: pJYDN3
FEATURES             Location/Qualifiers
     source          1..5254
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(194..868)
                     /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(869..1149)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(1170..1189)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1250..1268)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    1309..1812
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        1849..1953
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1954..2769
                     /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             1954..2769
                     /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(2027..2046)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2940..3528
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3429..3448
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3711..4152
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    3711..3828
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     4123..4146
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             4274..4312
                     /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"
     CDS             4654..4665
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             4666..4692
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4759..4788
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 tcccactgta cttttagctc gtacaaaata caatatactt ttcatttctc cgtaaacaac
       61 atgttttccc atgtaatatc cttttctatt tttcgttccg ttaccaactt tacacatact
      121 tttgcggtat ttcacaccgc atagatcggc aagtgcacaa acaatactta aataaatact
      181 actcagtaat aacctatttc ttagcatttt tgacgaaatt tgctattttg ttagagtctt
      241 ttacaccatt tgtctccaca cctccgctta catcaacacc aataacgcca tttaatctaa
      301 gcgcatcacc aacattttct ggcgtcagtc caccagctaa cataaaatgt aagctttcgg
      361 ggctctcttg ccttccaacc cagtcagaaa tcgagttcca atccaaaagt tcacctgtcc
      421 cacctgcttc tgaatcaaac aagggaataa acgaatgagg tttctgtgaa gctgcactga
      481 gtagtatgtt gcagtctttt ggaaatacga gtcttttaat aactggcaaa ccgaggaact
      541 cttggtattc ttgccacgac tcatctccat gcagttggac gatatcaatg ccgtaatcat
      601 tgaccagagc caaaacatcc tccttaggtt gattacgaaa cacgccaacc aagtatttcg
      661 gagtgcctga actattttta tatgctttta caagacttga aattttcctt gcaataaccg
      721 ggtcaattgt tctctttcta ttgggcacac atataatacc cagcaagtca gcatcggaat
      781 ctagagcaca ttctgcggcc tctgtgctct gcaagccgca aactttcacc aatggaccag
      841 aactacctgt gaaattaata acagacatac tccaagctgc ctttgtgtgc ttaatcacgt
      901 atactcacgt gctcaatagt caccaatgcc ctccctcttg gccctctcct tttctttttt
      961 cgaccgaatt aattcttaat cggcaaaaaa agaaaagctc cggatcaaga ttgtacgtaa
     1021 ggtgacaagc tatttttcaa taaagaatat cttccactac tgccatctgg cgtcataact
     1081 gcaaagtaca catatattac gatgctgttc tattaaatgc ttcctatatt atatatatag
     1141 taatgtcgtg atctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     1201 cagccccgac acccgccaac acccgctgac gcgcccttga gacgaaaggg cctcgtgata
     1261 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgga tcgcttgcct
     1321 gtaacttaca cgcgcctcgt atcttttaat gatggaataa tttgggaatt tactctgtgt
     1381 ttatttattt ttatgttttg tatttggatt ttagaaagta aataaagaag gtagaagagt
     1441 tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa aagcgtactt
     1501 tacatatata tttattagac aagaaaagca gattaaatag atatacattc gattaacgat
     1561 aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt ctacacagac aagatgaaac
     1621 aattcggcat taatacctga gagcaggaag agcaagataa aaggtagtat ttgttggcga
     1681 tccccctaga gtcttttaca tcttcggaaa acaaaaacta ttttttcttt aatttctttt
     1741 tttactttct atttttaatt tatatattta tattaaaaaa tttaaattat aattattttt
     1801 atagcacgtg atgaaaagga cccaggtggc acttttcggg gaaatgtgcg cggaacccct
     1861 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1921 taaatgcttc aataatattg aaaaaggaag agtatgagcc atattcaacg ggaaacgtct
     1981 tgctcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     2041 cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg
     2101 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     2161 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     2221 actcctgatg atgcatggtt actcaccact gcgatccccg ggaaaacagc attccaggta
     2281 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     2341 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     2401 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     2461 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataagctttt gccattctca
     2521 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     2581 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     2641 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     2701 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     2761 tttttctaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     2821 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     2881 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2941 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3001 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3061 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3121 aagaactctg tagcaccacc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3181 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3241 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3301 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3361 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3421 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3481 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3541 gcggcacttt gtaactgagc tgtcattttg ctggcccagc tggtcgaccg cggctaagca
     3601 gtactggtac cgttgtggaa atgtaaagag ccccattatc ttagcctaaa aaaaccttct
     3661 ctttggaact ttcagtaata cgcttaactg ctcattgcta tattgaagta cggattagaa
     3721 gccgccgagc gggtgacagc cctccgaagg aagactctcc tccgtgcgtc ctcgtcttca
     3781 ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa caataaagat
     3841 tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc tggccccaca
     3901 aaccttcaaa tgaacgaatc aaattaacaa ccataggatg ataatgcgat tagtttttta
     3961 gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt aacagatata
     4021 taaatgcaaa aactgcataa ccactttaac taatactttc aacattttcg gtttgtatta
     4081 cttcttattc aaatgtaata aaagtatcaa caaaaaattg ttaatatacc tctatacttt
     4141 aacgtcaagg agaaaaaacc ccggatcgaa ttccctactt catacatttt caattaagat
     4201 gaggttccca tctattttca ccgctgttgt ttttgctgct tcttctgctt tggctgctcc
     4261 agctaatggt ccatccagat tggaagaaga attgagaaga agattaaccg aaccacatat
     4321 ggaagcataa taattaaggc gccttatctc gaggggggcg gatcctctgg tggaggtggt
     4381 tcgggcggag gtggtaacgg atctaatggt tctggcggta gctctgcaca ggaactgaca
     4441 actatatgcg agcaaatccc ctcaccaact ttagaatcga cgccgtactc tttgtcaacg
     4501 actactattt tggccaacgg gaaggcaatg caaggagttt ttgaatatta caaatcagta
     4561 acgtttgtca gtaattgcgg ttctcacccc tcaacaacta gcaaaggcag ccccataaac
     4621 acacagtatg tttttaagga caatagctcg acgattgaag gtagataccc atacgacgtt
     4681 ccagactacg ctctgcaggc tagtggtgga ggaggctctg gtggaggtgg tagcggaggc
     4741 ggagggtcgg ctagccatga acagaagttg atctctgaag aagatttgat gttagaaaaa
     4801 tttgttggca cctggaagat cgaatcctct gaaaattttg gtgaatactt gaaggctatc
     4861 ggtgccccaa aagaattggc tgatgctggt gatgctacta ctccagtctt gtacatttct
     4921 caaaaggatg gtgataagat gaccgtcaag attgaaaacg gtccaccaac ttttttggat
     4981 acccaagttt ctttcaagtt gggtgaagaa ttcgacgaat ttccatccga tagaagaaag
     5041 ggtgttaagt ccgttgttaa cttgtctggt gagaagttgg tttacgttca aaagtgggat
     5101 ggtaaagaaa ccacttacgt cagagaaatc aaggacggta aattggttgt tactttgacc
     5161 atgggtgatg ttgttgctgt tagatcttat agaagggcct ctgaataata gagatctgat
     5221 aacaacagtg tagatgtaac aaaatcgact ttgt
//
