LOCUS       sequence-321464-        5255 bp    DNA     circular SYN 14-APR-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 5255)
  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 5255)
  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: pJYDN3
FEATURES             Location/Qualifiers
     source          1..5255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(199..873)
                     /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(874..1154)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(1175..1194)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1255..1273)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    1314..1818
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        1855..1959
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1960..2775
                     /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             1960..2775
                     /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(2033..2052)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2946..3534
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3435..3454
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3717..4158
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    3717..3834
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     4129..4152
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             4280..4318
                     /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             4660..4671
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             4672..4698
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4765..4794
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 tttgttccca ctgtactttt agctcgtaca aaatacaata tacttttcat ttctccgtaa
       61 acaacatgtt ttcccatgta atatcctttt ctatttttcg ttccgttacc aactttacac
      121 atacttttgc ggtatttcac accgcataga tcggcaagtg cacaaacaat acttaaataa
      181 atactactca gtaataacct atttcttagc atttttgacg aaatttgcta ttttgttaga
      241 gtcttttaca ccatttgtct ccacacctcc gcttacatca acaccaataa cgccatttaa
      301 tctaagcgca tcaccaacat tttctggcgt cagtccacca gctaacataa aatgtaagct
      361 ttcggggctc tcttgccttc caacccagtc agaaatcgag ttccaatcca aaagttcacc
      421 tgtcccacct gcttctgaat caaacaaggg aataaacgaa tgaggtttct gtgaagctgc
      481 actgagtagt atgttgcagt cttttggaaa tacgagtctt ttaataactg gcaaaccgag
      541 gaactcttgg tattcttgcc acgactcatc tccatgcagt tggacgatat caatgccgta
      601 atcattgacc agagccaaaa catcctcctt aggttgatta cgaaacacgc caaccaagta
      661 tttcggagtg cctgaactat ttttatatgc ttttacaaga cttgaaattt tccttgcaat
      721 aaccgggtca attgttctct ttctattggg cacacatata atacccagca agtcagcatc
      781 ggaatctaga gcacattctg cggcctctgt gctctgcaag ccgcaaactt tcaccaatgg
      841 accagaacta cctgtgaaat taataacaga catactccaa gctgcctttg tgtgcttaat
      901 cacgtatact cacgtgctca atagtcacca atgccctccc tcttggccct ctccttttct
      961 tttttcgacc gaattaattc ttaatcggca aaaaaagaaa agctccggat caagattgta
     1021 cgtaaggtga caagctattt ttcaataaag aatatcttcc actactgcca tctggcgtca
     1081 taactgcaaa gtacacatat attacgatgc tgttctatta aatgcttcct atattatata
     1141 tatagtaatg tcgtgatcta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     1201 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctcgagacga aagggcctcg
     1261 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acggatcgct
     1321 tgcctgtaac ttacacgcgc ctcgtatctt ttaatgatgg aataatttgg gaatttactc
     1381 tgtgtttatt tatttttatg ttttgtattt ggattttaga aagtaaataa agaaggtaga
     1441 agagttacgg aatgaagaaa aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc
     1501 gtactttaca tatatattta ttagacaaga aaagcagatt aaatagatat acattcgatt
     1561 aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac acagacaaga
     1621 tgaaacaatt cggcattaat acctgagagc aggaagagca agataaaagg tagtatttgt
     1681 tggcgatccc cctagagtct tttacatctt cggaaaacaa aaactatttt ttctttaatt
     1741 tcttttttta ctttctattt ttaatttata tatttatatt aaaaaattta aattataatt
     1801 atttttatag cacgtgatga aaaggaccca ggtggcactt ttcggggaaa tgtgcgcgga
     1861 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1921 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagccatat tcaacgggaa
     1981 acgtcttgct cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     2041 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc
     2101 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     2161 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     2221 atccgtactc ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc
     2281 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     2341 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     2401 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     2461 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa gcttttgcca
     2521 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     2581 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     2641 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     2701 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     2761 gatgagtttt tctaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2821 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2881 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2941 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3001 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3061 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3121 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3181 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3241 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3301 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3361 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3421 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3481 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg aaaaaacgcc
     3541 agcaacgcgg cactttgtaa ctgagctgtc attttgctgg cccagctggt cgaccgcggc
     3601 taagcagtac tggtaccgtt gtggaaatgt aaagagcccc attatcttag cctaaaaaaa
     3661 ccttctcttt ggaactttca gtaatacgct taactgctca ttgctatatt gaagtacgga
     3721 ttagaagccg ccgagcgggt gacagccctc cgaaggaaga ctctcctccg tgcgtcctcg
     3781 tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctcgc gccgcactgc tccgaacaat
     3841 aaagattcta caatactagc ttttatggtt atgaagagga aaaattggca gtaacctggc
     3901 cccacaaacc ttcaaatgaa cgaatcaaat taacaaccat aggatgataa tgcgattagt
     3961 tttttagcct tatttctggg gtaattaatc agcgaagcga tgatttttga tctattaaca
     4021 gatatataaa tgcaaaaact gcataaccac tttaactaat actttcaaca ttttcggttt
     4081 gtattacttc ttattcaaat gtaataaaag tatcaacaaa aaattgttaa tatacctcta
     4141 tactttaacg tcaaggagaa aaaaccccgg atcgaattcc ctacttcata cattttcaat
     4201 taagatgagg ttcccatcta ttttcaccgc tgttgttttt gctgcttctt ctgctttggc
     4261 tgctccagct aatggtccat ccagattgga agaagaattg agaagaagat taaccgaacc
     4321 acatatggaa gcataataat taaggcgcct tatctcgagg ggggcggatc ctctggtgga
     4381 ggtggttcgg gcggaggtgg taacggatct aatggttctg gcggtagctc tgcacaggaa
     4441 ctgacaacta tatgcgagca aatcccctca ccaactttag aatcgacgcc gtactctttg
     4501 tcaacgacta ctattttggc caacgggaag gcaatgcaag gagtttttga atattacaaa
     4561 tcagtaacgt ttgtcagtaa ttgcggttct cacccctcaa caactagcaa aggcagcccc
     4621 ataaacacac agtatgtttt taaggacaat agctcgacga ttgaaggtag atacccatac
     4681 gacgttccag actacgctct gcaggctagt ggtggaggag gctctggtgg aggtggtagc
     4741 ggaggcggag ggtcggctag ccatgaacag aagttgatct ctgaagaaga tttgatgtta
     4801 gaaaaatttg ttggcacctg gaagatcgaa tcctctgaaa attttggtga atacttgaag
     4861 gctatcggtg ccccaaaaga attggctgat gctggtgatg ctactactcc agtcttgtac
     4921 atttctcaaa aggatggtga taagatgacc gtcaagattg aaaacggtcc accaactttt
     4981 ttggataccc aagtttcttt caagttgggt gaagaattcg acgaatttcc atccgataga
     5041 agaaagggtg ttaagtccgt tgttaacttg tctggtgaga agttggttta cgttcaaaag
     5101 tgggatggta aagaaaccac ttacgtcaga gaaatcaagg acggtaaatt ggttgttact
     5161 ttgaccatgg gtgatgttgt tgctgttaga tcttatagaa gggcctctga ataatagaga
     5221 tctgataaca acagtgtaga tgtaacaaaa tcgac
//
