LOCUS       sequence-487916-        4164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dropout vector for genomic integration into yeast strain LFYalpha. 
            GFP is flanked by BamHI sites for easy cloning using Gibson 
            assembly. Construct contains attB site for the serine recombinase 
            BxB1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4164)
  AUTHORS   Lester Frei, Sai Tosa Reddy
  TITLE     A yeast mating-based platform enables the generation and screening 
            of ultra large antibody libraries
  JOURNAL   bioRxiv
REFERENCE   2  (bases 1 to 4164)
  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"
COMMENT     Sequence Label: pLFYalpha
FEATURES             Location/Qualifiers
     source          1..4164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     115..134
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     199..218
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      432..1020
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     921..940
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        complement(1339..1440)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     promoter        1641..2082
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1641..1758
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2053..2076
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     RBS             2286..2297
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2304..3017
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      3029..3100
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3116..3143
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    3163..3212
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     CDS             complement(join(3556..4164,1..207))
                     /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(join(3556..4164,1..207))
                     /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"
ORIGIN
        1 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
       61 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
      121 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctggagca
      181 agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga
      241 cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg
      301 agacacaacg tggctttgtt gaataaatcg aacttttgct gagttgaagg atcagtcatg
      361 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
      421 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
      481 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      541 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
      601 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      661 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      721 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      781 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      841 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      901 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      961 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     1021 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     1081 ttctttcctg cgttatcccc tgattctgtg gataaccgtc aagtatttcg gagtgcctga
     1141 actattttta tatgctttta caagacttga aattttcctt gcaataaccg ggtcaattgt
     1201 tctctttcta ttgggcacac atataatacc cagcaagtca gcatcggaat ctagagcaca
     1261 ttctgcggcc tctgtgctct gcaagccgca aactttcacc aatggaccag aactacctgt
     1321 gaaattaata acagacatac tccaagctgc ctttgtgtgc ttaatcacgt atactcacgt
     1381 gctcaatagt caccaatgcc ctccctcttg gccctctcct tttctttttt cgaccgaatt
     1441 atacgtaaga gctcaaggca ggtcaattcg cactgtcagg gtcacatggg tgtttggcac
     1501 taccgacacg aacctcagta agcgttcttc ctaccagagg tctgtggcat cgtggtcaaa
     1561 agtgcggatt tcgtatttgc tgctcgtcta tactttcaca atcttgacct gcacggcaaa
     1621 gagacgcttc ttgtggagct cggattagaa gccgccgagc gggtgacagc cctccgaagg
     1681 aagactctcc tccgtgcgtc ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc
     1741 tcgcgccgca ctgctccgaa caataaagat tctacaatac tagcttttat ggttatgaag
     1801 aggaaaaatt ggcagtaacc tggccccaca aaccttcaaa tgaacgaatc aaattaacaa
     1861 ccataggatg ataatgcgat tagtttttta gccttatttc tggggtaatt aatcagcgaa
     1921 gcgatgattt ttgatctatt aacagatata taaatgcaaa aactgcataa ccactttaac
     1981 taatactttc aacattttcg gtttgtatta cttcttattc aaatgtaata aaagtatcaa
     2041 caaaaaattg ttaatatacc tctatacttt aacgtcaagg agaaaaaacc ccggatcgaa
     2101 ttcagctaaa aaaaccaaaa aatttcagat gggatccgaa agtgaaacgt gatttcatgc
     2161 gtcattttga acattttgta aatcttattt aataatgtgt gcggcaattc acatttaatt
     2221 tatgaatgtt ttcttaacat cgcggcaact caagaaacgg caggttcgga tcttagctac
     2281 tagagaaaga ggagaaatac tagatgcgta aaggcgaaga gctgttcact ggtgtcgtcc
     2341 ctattctggt ggaactggat ggtgatgtca acggtcataa gttttccgtg cgtggcgagg
     2401 gtgaaggtga cgcaactaat ggtaaactga cgctgaagtt catctgtact actggtaaac
     2461 tgccggttcc ttggccgact ctggtaacga cgctgactta tggtgttcag tgctttgctc
     2521 gttatccgga ccatatgaag cagcatgact tcttcaagtc cgccatgccg gaaggctatg
     2581 tgcaggaacg cacgatttcc tttaaggatg acggcacgta caaaacgcgt gcggaagtga
     2641 aatttgaagg cgataccctg gtaaaccgca ttgagctgaa aggcattgac tttaaagagg
     2701 acggcaatat cctgggccat aagctggaat acaattttaa cagccacaat gtttacatca
     2761 ccgccgataa acaaaaaaat ggcattaaag cgaattttaa aattcgccac aacgtggagg
     2821 atggcagcgt gcagctggct gatcactacc agcaaaacac tccaatcggt gatggtcctg
     2881 ttctgctgcc agacaatcac tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg
     2941 agaaacgcga tcatatggtt ctgctggagt tcgtaaccgc agcgggcatc acgcatggta
     3001 tggatgaact gtacaaatga ccaggcatca aataaaacga aaggctcagt cgaaagactg
     3061 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc
     3121 accttcgggt gggcctttct gcgtttataa ggatcctaat aatcggccgg cttgtcgacg
     3181 acggcggtct ccgtcgtcag gatcatccgg gctagctcga gatctgataa caacagtgta
     3241 gatgtaacaa aatcgacttt gttcccactg tacttttagc tcgtacaaaa tacaatatac
     3301 ttttcatttc tccgtaaaca acatgttttc ccatgtaata tccttttcta tttttcgttc
     3361 cgttaccaac tttacacata ctttatatag ctattcactt ctatacacta aaaaactaag
     3421 acaattttaa ttttgctgcc tgccatattt caatttgtta taaattccta taatttatcc
     3481 tattagtagc taaaaaaaga tgaatgtgaa tcgaatccta agagaattgt gttacaacca
     3541 attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     3601 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     3661 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     3721 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     3781 accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac
     3841 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     3901 attcattcgt gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa aaggacaatt
     3961 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     4021 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     4081 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     4141 ttccgtcagc cagtttagtc tgac
//
