LOCUS       sequence-321460-        5727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Positive control plasmid for pJYDNg carrying the eUnaG2 bilirubin 
            dependent fluorescent protein fused to the C-terminus of Aga2p 
            protein. This plasmid contains a long 2G linker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5727)
  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 5727)
  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: pJYDNgp
FEATURES             Location/Qualifiers
     source          1..5727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        90..194
                     /gene="bla"
                     /label=AmpR promoter
     CDS             195..1010
                     /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             195..1010
                     /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(268..287)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1181..1769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1670..1689
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1924..1941
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2058..2079
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2094..2099,2100..2117,2118..2124)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2132..2148
                     /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)."
     primer_bind     2137..2159
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2156..2172
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2156..2172
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2193..2211
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2241..2893
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    2445..2562
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2857..2880
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             3365..3376
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3377..3403
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3470..3499
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(4159..4833)
                     /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(4834..5114)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(5135..5154)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5215..5233)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    join(5274..5727,1..53)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 ttatatattt atattaaaaa atttaaatta taattatttt tatagcacgt gatgaaaagg
       61 acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
      121 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
      181 gaaaaaggaa gagtatgagc catattcaac gggaaacgtc ttgctcgagg ccgcgattaa
      241 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
      301 caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac
      361 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
      421 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
      481 tactcaccac tgcgatcccc gggaaaacag cattccaggt attagaagaa tatcctgatt
      541 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
      601 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
      661 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
      721 aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca gtcgtcactc
      781 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
      841 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
      901 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
      961 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa ctgtcagacc
     1021 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1081 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1141 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1201 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1261 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1321 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     1381 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     1441 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     1501 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     1561 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     1621 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     1681 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     1741 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggcactt tgtaactgag
     1801 ctgtcatttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     1861 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     1921 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     1981 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     2041 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     2101 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     2161 aacagctatg accatgatta cgccaagctc gaaattaacc ctcactaaag ggaacaaaag
     2221 ctggtaccaa ttccttgaat tttcaaaaat tcttactttt tttttggatg gacgcaaaga
     2281 agtttaataa tcatattaca tggcattacc accatataca tatccatatc taatcttact
     2341 tatatgttgt ggaaatgtaa agagccccat tatcttagcc taaaaaaacc ttctctttgg
     2401 aactttcagt aatacgctta actgctcatt gctatattga agtacggatt agaagccgcc
     2461 gagcgggtga cagccctccg aaggaagact ctcctccgtg cgtcctcgtc ttcaccggtc
     2521 gcgttcctga aacgcagatg tgcctcgcgc cgcactgctc cgaacaataa agattctaca
     2581 atactagctt ttatggttat gaagaggaaa aattggcagt aacctggccc cacaaacctt
     2641 caaatgaacg aatcaaatta acaaccatag gatgataatg cgattagttt tttagcctta
     2701 tttctggggt aattaatcag cgaagcgatg atttttgatc tattaacaga tatataaatg
     2761 caaaaactgc ataaccactt taactaatac tttcaacatt ttcggtttgt attacttctt
     2821 attcaaatgt aataaaagta tcaacaaaaa attgttaata tacctctata ctttaacgtc
     2881 aaggagaaaa aaccccggat cgaattccct acttcataca ttttcaatta agatgaggtt
     2941 cccatctatt ttcaccgctg ttgtttttgc tgcttcttct gctttggctg ctccagctaa
     3001 tggtcatatg gaagcatgct taattaaggc gccttatctc gaggggggcg gatccttctc
     3061 ccaaaagttg gacatcaact tgttggataa cgttgtcaac tcttcatatc atggtgaagg
     3121 tgtttctgga ggttctgcac aggaactgac aactatatgc gagcaaatcc cctcaccaac
     3181 tttagaatcg acgccgtact ctttgtcaac gactactatt ttggccaacg ggaaggcaat
     3241 gcaaggagtt tttgaatatt acaaatcagt aacgtttgtc agtaattgcg gttctcaccc
     3301 ctcaacaact agcaaaggca gccccataaa cacacagtat gtttttaagg acaatagctc
     3361 gacgattgaa ggtagatacc catacgacgt tccagactac gctctgcagg ctagtggtgg
     3421 aggaggctct ggtggaggtg gtagcggagg cggagggtcg gctagccatg aacagaagtt
     3481 gatctctgaa gaagatttga tgttagaaaa atttgttggc acctggaaga tcgaatcctc
     3541 tgaaaatttt ggtgaatact tgaaggctat cggtgcccca aaagaattgg ctgatgctgg
     3601 tgatgctact actccagtct tgtacatttc tcaaaaggat ggtgataaga tgaccgtcaa
     3661 gattgaaaac ggtccaccaa cttttttgga tacccaagtt tctttcaagt tgggtgaaga
     3721 attcgacgaa tttccatccg atagaagaaa gggtgttaag tccgttgtta acttgtctgg
     3781 tgagaagttg gtttacgttc aaaagtggga tggtaaagaa accacttacg tcagagaaat
     3841 caaggacggt aaattggttg ttactttgac catgggtgat gttgttgctg ttagatctta
     3901 tagaagggcc tctgaataat agagatctga taacaacagt gtagatgtaa caaaatcgac
     3961 tttgttccca ctgtactttt agctcgtaca aaatacaata tacttttcat ttctccgtaa
     4021 wcaacatgtt ttcccatgta atatcctttt ctatttttcg ttccgttacc aactttacac
     4081 atacttttgc ggtatttcac accgcataga tcggcaagtg cacaaacaat acttaaataa
     4141 atactactca gtaataacct atttcttagc atttttgacg aaatttgcta ttttgttaga
     4201 gtcttttaca ccatttgtct ccacacctcc gcttacatca acaccaataa cgccatttaa
     4261 tctaagcgca tcaccaacat tttctggcgt cagtccacca gctaacataa aatgtaagct
     4321 ttcggggctc tcttgccttc caacccagtc agaaatcgag ttccaatcca aaagttcacc
     4381 tgtcccacct gcttctgaat caaacaaggg aataaacgaa tgaggtttct gtgaagctgc
     4441 actgagtagt atgttgcagt cttttggaaa tacgagtctt ttaataactg gcaaaccgag
     4501 gaactcttgg tattcttgcc acgactcatc tccatgcagt tggacgatat caatgccgta
     4561 atcattgacc agagccaaaa catcctcctt aggttgatta cgaaacacgc caaccaagta
     4621 tttcggagtg cctgaactat ttttatatgc ttttacaaga cttgaaattt tccttgcaat
     4681 aaccgggtca attgttctct ttctattggg cacacatata atacccagca agtcagcatc
     4741 ggaatctaga gcacattctg cggcctctgt gctctgcaag ccgcaaactt tcaccaatgg
     4801 accagaacta cctgtgaaat taataacaga catactccaa gctgcctttg tgtgcttaat
     4861 cacgtatact cacgtgctca atagtcacca atgccctccc tcttggccct ctccttttct
     4921 tttttcgacc gaattaattc ttaatcggca aaaaaagaaa agctccggat caagattgta
     4981 cgtaaggtga caagctattt ttcaataaag aatatcttcc actactgcca tctggcgtca
     5041 taactgcaaa gtacacatat attacgatgc tgttctatta aatgcttcct atattatata
     5101 tatagtaatg tcgtgatcta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     5161 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctcgagacga aagggcctcg
     5221 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acggatcgct
     5281 tgcctgtaac ttacacgcgc ctcgtatctt ttaatgatgg aataatttgg gaatttactc
     5341 tgtgtttatt tatttttatg ttttgtattt ggattttaga aagtaaataa agaaggtaga
     5401 agagttacgg aatgaagnaa aaaaaaaaat aaacaaaggt ttaaaaaatt tcaacaaaaa
     5461 gcgtacttta catatatatt tattagacaa gaaaagcaga ttaaatagat atacattcga
     5521 ttaacgataa gtaaaatgta aaatcacagg attttcgtgt gtggtcttct acacagacaa
     5581 gatgaaacaa ttcggcatta atacctgaga gcaggaagag caagataaaa ggtagtattt
     5641 gttggcgatc cccctagagt cttttacatc ttcggaaaac aaaaactatt ttttctttaa
     5701 tttctttttt tactttctat ttttaat
//
