LOCUS       sequence-321472-        5694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Positive control plasmid for pJYDN3n plasmid expression attempts. It
            contains both eUnaG2 bilirubin dependent fluorescent protein and 
            ALFA-tag for DnbALFA based detection on the cell surface..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5694)
  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 5694)
  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: pJYDN3p
FEATURES             Location/Qualifiers
     source          1..5694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     488..506
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     567..586
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        607..887
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             888..1562
                     /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"
     CDS             complement(2222..2251)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(2318..2344)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(2345..2356)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(2645..2683)
                     /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"
     promoter        2798..3450
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     primer_bind     complement(2811..2834)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    3129..3246
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     promoter        complement(3480..3498)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3519..3535)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3519..3535)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3532..3554)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3543..3559
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3567..3573,3574..3591,3592..3597))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3612..3633
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3750..3767)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3922..4510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4002..4021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4681..5496)
                     /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             complement(4681..5496)
                     /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     5404..5423
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5497..5601)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(5638..5694,1..447)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 aaaatagaaa gtaaaaaaag aaattaaaga aaaaatagtt tttgttttcc gaagatgtaa
       61 aagactctag ggggatcgcc aacaaatact accttttatc ttgctcttcc tgctctcagg
      121 tattaatgcc gaattgtttc atcttgtctg tgtagaagac cacacacgaa aatcctgtga
      181 ttttacattt tacttatcgt taatcgaatg tatatctatt taatctgctt ttcttgtcta
      241 ataaatatat atgtaaagta cgctttttgt tgaaattttt taaacctttg tttatttttt
      301 tttcttcatt ccgtaactct tctaccttct ttatttactt tctaaaatcc aaatacaaaa
      361 cataaaaata aataaacaca gagtaaattc ccaaattatt ccatcattaa aagatacgag
      421 gcgcgtgtaa gttacaggca agcgatccgt ctaagaaacc attattatca tgacattaac
      481 ctataaaaat aggcgtatca cgaggccctt tcgtctcgag ggcgcgtcag cgggtgttgg
      541 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
      601 tagatcacga cattactata tatataatat aggaagcatt taatagaaca gcatcgtaat
      661 atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa
      721 aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa
      781 gaattaattc ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat
      841 tgagcacgtg agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta
      901 atttcacagg tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg
      961 cagaatgtgc tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa
     1021 agagaacaat tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa
     1081 atagttcagg cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg
     1141 ttttggctct ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt
     1201 ggcaagaata ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag
     1261 actgcaacat actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg
     1321 attcagaagc aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg
     1381 gaaggcaaga gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa
     1441 atgttggtga tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg
     1501 agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat
     1561 aggttattac tgagtagtat ttatttaagt attgtttgtg cacttgccga tctatgcggt
     1621 gtgaaatacc gcaaaagtat gtgtaaagtt ggtaacggaa cgaaaaatag aaaaggatat
     1681 tacatgggaa aacatgttgt ttacggagaa atgaaaagta tattgtattt tgtacgagct
     1741 aaaagtacag tgggaacaaa gtcgattttg ttacatctac actgttgtta tcagatctct
     1801 attattcaga ggcccttcta taagatctaa cagcaacaac atcacccatg gtcaaagtaa
     1861 caaccaattt accgtccttg atttctctga cgtaagtggt ttctttacca tcccactttt
     1921 gaacgtaaac caacttctca ccagacaagt taacaacgga cttaacaccc tttcttctat
     1981 cggatggaaa ttcgtcgaat tcttcaccca acttgaaaga aacttgggta tccaaaaaag
     2041 ttggtggacc gttttcaatc ttgacggtca tcttatcacc atccttttga gaaatgtaca
     2101 agactggagt agtagcatca ccagcatcag ccaattcttt tggggcaccg atagccttca
     2161 agtattcacc aaaattttca gaggattcga tcttccaggt gccaacaaat ttttctaaca
     2221 tcaaatcttc ttcagagatc aacttctgtt catggctagc cgaccctccg cctccgctac
     2281 cacctccacc agagcctcct ccaccactag cctgcagagc gtagtctgga acgtcgtatg
     2341 ggtatctacc ttcaatcgtc gagctattgt ccttaaaaac atactgtgtg tttatggggc
     2401 tgcctttgct agttgttgag gggtgagaac cgcaattact gacaaacgtt actgatttgt
     2461 aatattcaaa aactccttgc attgccttcc cgttggccaa aatagtagtc gttgacaatg
     2521 agtacggcgt cgattctaaa gttggtgagg ggatttgctc gcatatagtt gtcagttcct
     2581 gtgcagaaac ggatccgccc ccctcgagat aaggcgcctt aattaagcat gcttccatat
     2641 gtggttcggt taatcttctt ctcaattctt cttccaatct ggatggacca ttagctggag
     2701 cagccaaagc agaagaagca gcaaaaacaa cagcggtgaa aatagatggg aacctcatct
     2761 taattgaaaa tgtatgaagt agggaattcg atccggggtt ttttctcctt gacgttaaag
     2821 tatagaggta tattaacaat tttttgttga tacttttatt acatttgaat aagaagtaat
     2881 acaaaccgaa aatgttgaaa gtattagtta aagtggttat gcagtttttg catttatata
     2941 tctgttaata gatcaaaaat catcgcttcg ctgattaatt accccagaaa taaggctaaa
     3001 aaactaatcg cattatcatc ctatggttgt taatttgatt cgttcatttg aaggtttgtg
     3061 gggccaggtt actgccaatt tttcctcttc ataaccataa aagctagtat tgtagaatct
     3121 ttattgttcg gagcagtgcg gcgcgaggca catctgcgtt tcaggaacgc gaccggtgaa
     3181 gacgaggacg cacggaggag agtcttcctt cggagggctg tcacccgctc ggcggcttct
     3241 aatccgtact tcaatatagc aatgagcagt taagcgtatt actgaaagtt ccaaagagaa
     3301 ggttttttta ggctaagata atggggctct ttacatttcc acaacatata agtaagatta
     3361 gatatggata tgtatatggt ggtaatgcca tgtaatatga ttattaaact tctttgcgtc
     3421 catccaaaaa aaaagtaaga atttttgaaa attcaaggaa ttggtaccag cttttgttcc
     3481 ctttagtgag ggttaatttc gagcttggcg taatcatggt catagctgtt tcctgtgtga
     3541 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3601 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3661 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3721 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3781 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3841 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaatgacag ctcagttaca
     3901 aagtgccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3961 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4021 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4081 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4141 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4201 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4261 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4321 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4381 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4441 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4501 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4561 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4621 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4681 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     4741 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     4801 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     4861 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     4921 tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca
     4981 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     5041 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     5101 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     5161 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     5221 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     5281 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     5341 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     5401 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     5461 cctcgagcaa gacgtttccc gttgaatatg gctcatactc ttcctttttc aatattattg
     5521 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5581 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt ccttttcatc
     5641 acgtgctata aaaataatta taatttaaat tttttaatat aaatatataa atta
//
