LOCUS       sequence-430578-        2775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stuffer plasmid with golden gate compatible bidirectional promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2775)
  AUTHORS   Petersen BM, Kirby MB, Chrispens KM, Irvin OM, Strawn IK, Haas CM, 
            Walker AM, Baumer ZT, Ulmer SA, Ayala E, Rhodes ER, Guthmiller JJ, 
            Steiner PJ, Whitehead TA
  TITLE     An integrated technology for quantitative wide mutational scanning 
            of human antibody Fab libraries.
  JOURNAL   Nat Commun. 2024 May 10;15(1):3974. doi: 10.1038/s41467-024-48072-z.
  PUBMED    38730230
REFERENCE   2  (bases 1 to 2775)
  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: "Nat 
            Commun."; date: "2024-05-10"; volume: "15(1):3974. doi"; pages: " 
            10.1038/s41467-024-48072-z"
COMMENT     Sequence Label: pBDP
FEATURES             Location/Qualifiers
     source          1..2775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..760
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     primer_bind     complement(109..132)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    427..544
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             complement(1068..1883)
                     /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(1068..1883)
                     /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     1791..1810
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1875..1894
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2108..2696
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2597..2616
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgattctgt ggataaccgt gcggccgcgc ggtctcaacc tgctaaaacg ctagcaataa
       61 cagaaaatat tgaaaaacag cgaagtaact gcatggtttt ttctccttga cgttaaagta
      121 tagaggtata ttaacaattt tttgttgata cttttattac atttgaataa gaagtaatac
      181 aaaccgaaaa tgttgaaagt attagttaaa gtggttatgc agtttttgca tttatatatc
      241 tgttaataga tcaaaaatca tcgcttcgct gattaattac cccagaaata aggctaaaaa
      301 actaatcgca ttatcatcct atggttgtta atttgattcg ttcatttgaa ggtttgtggg
      361 gccaggttac tgccaatttt tcctcttcat aaccataaaa gctagtattg tagaatcttt
      421 attgttcgga gcagtgcggc gcgaggcaca tctgcgtttc aggaacgcga ccggtgaata
      481 cgaggacgca cggaggagaa tcttccttcg gagggctgtc acccgctcgg cggcttctaa
      541 tccgtacttc aatatagcaa tgagcagtta agcgtattac tgaaagttcc aaagagaagg
      601 tttttttagg ctaagataat ggggctcttt acatttccac aacatataag taagattaga
      661 tatggatatg tatatggata tgtatatggt ggtaatgcca tgtaatatga ttattaaact
      721 tctttgcgtc catccaaaaa aaaagtaaga atttttgaaa attcaccatg agatttcctt
      781 caatttttac tgcagtttta ttcgcagcat cctccgcatt agctgctcca gctaacacta
      841 caacagaaga tgaaacggca caaattccgg ctgaagctgt catcgattac tcagatttag
      901 aaggggattt cgatgctgct gctttgccat tgtccaacag cacaaataac gggttatcct
      961 ccacaaatac tactattgcc agcattgctg ctaaagaaga aggggtacaa ctcgattgag
     1021 accccgcggc cgccctcagc gtgttacaac caattaacca attctgatta gaaaaactca
     1081 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     1141 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     1201 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     1261 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     1321 aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     1381 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgagg
     1441 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     1501 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     1561 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     1621 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     1681 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     1741 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     1801 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct ggagcaagac
     1861 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
     1921 tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac
     1981 acaacgtggc tttgttgaat aaatcgaact tttgctgagt tgaaggatca gtcatgacca
     2041 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     2101 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     2161 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2221 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     2281 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2341 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2401 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2461 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     2521 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     2581 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2641 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     2701 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     2761 ttcctgcgtt atccc
//
