LOCUS       sequence-487915-        4168 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Dropout vector for genomic integration into yeast strain LFYa. GFP 
            is flanked by BamHI sites for easy cloning using Gibson assembly. 
            Construct contains the attP site for the serine recombinase BxB1. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4168)
  TITLE     Reddy lab plasmids 2025
REFERENCE   2  (bases 1 to 4168)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pLFYa
FEATURES             Location/Qualifiers
     source          1..4168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        610..1051
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    610..727
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     1022..1045
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     RBS             1255..1266
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1273..1986
                     /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      1998..2069
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2085..2112
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    2132..2189
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attP site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     CDS             complement(2533..3348)
                     /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(2533..3348)
                     /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     3256..3275
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3340..3359
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3573..4161
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4062..4081
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
       61 ctgcgttatc ccctgattct gtggataacc gtaactccag taattccttg gtggtacgaa
      121 catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat agcttggcag
      181 caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac atgatttatc
      241 ttcgtttcct gcacgttttt gttctgtgca gttgggttaa gaatactggg caatttcatg
      301 tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc ttccttcgtt
      361 cttccttctg ttcggagatt accgaatcaa aaaaatttca aggaaaccga cgtagccttg
      421 tagtcagtac gtagcctggt cgcataagta cagtagatcc tccctgcgca tcctatttat
      481 taagttaatt ctacagcaaa acgatcatat gcagatccgc agtggccggt agacacacgt
      541 ccaccccgct gctctgtgac agggactaaa gaggcgaaga ttatcgtgtg tgccccgtta
      601 tggtcgagtc ggattagaag ccgccgagcg ggtgacagcc ctccgaagga agactctcct
      661 ccgtgcgtcc tcgtcttcac cggtcgcgtt cctgaaacgc agatgtgcct cgcgccgcac
      721 tgctccgaac aataaagatt ctacaatact agcttttatg gttatgaaga ggaaaaattg
      781 gcagtaacct ggccccacaa accttcaaat gaacgaatca aattaacaac cataggatga
      841 taatgcgatt agttttttag ccttatttct ggggtaatta atcagcgaag cgatgatttt
      901 tgatctatta acagatatat aaatgcaaaa actgcataac cactttaact aatactttca
      961 acattttcgg tttgtattac ttcttattca aatgtaataa aagtatcaac aaaaaattgt
     1021 taatatacct ctatacttta acgtcaagga gaaaaaaccc cggatcgaat tcagctaaaa
     1081 aaaccaaaaa atttcagatg ggatccgaaa gtgaaacgtg atttcatgcg tcattttgaa
     1141 cattttgtaa atcttattta ataatgtgtg cggcaattca catttaattt atgaatgttt
     1201 tcttaacatc gcggcaactc aagaaacggc aggttcggat cttagctact agagaaagag
     1261 gagaaatact agatgcgtaa aggcgaagag ctgttcactg gtgtcgtccc tattctggtg
     1321 gaactggatg gtgatgtcaa cggtcataag ttttccgtgc gtggcgaggg tgaaggtgac
     1381 gcaactaatg gtaaactgac gctgaagttc atctgtacta ctggtaaact gccggttcct
     1441 tggccgactc tggtaacgac gctgacttat ggtgttcagt gctttgctcg ttatccggac
     1501 catatgaagc agcatgactt cttcaagtcc gccatgccgg aaggctatgt gcaggaacgc
     1561 acgatttcct ttaaggatga cggcacgtac aaaacgcgtg cggaagtgaa atttgaaggc
     1621 gataccctgg taaaccgcat tgagctgaaa ggcattgact ttaaagagga cggcaatatc
     1681 ctgggccata agctggaata caattttaac agccacaatg tttacatcac cgccgataaa
     1741 caaaaaaatg gcattaaagc gaattttaaa attcgccaca acgtggagga tggcagcgtg
     1801 cagctggctg atcactacca gcaaaacact ccaatcggtg atggtcctgt tctgctgcca
     1861 gacaatcact atctgagcac gcaaagcgtt ctgtctaaag atccgaacga gaaacgcgat
     1921 catatggttc tgctggagtt cgtaaccgca gcgggcatca cgcatggtat ggatgaactg
     1981 tacaaatgac caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     2041 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg
     2101 ggcctttctg cgtttataag gatcctaata agtcggggtt tgtaccgtac accactgaga
     2161 ccgcggtggt tgaccagaca aaccacgact agctcgagat ctgataacaa cagtgtagat
     2221 gtaacaaaat cgactttgtt cccactgtac ttttagctcg tacaaaatac aatatacttt
     2281 tcatttctcc gtaaacaaca tgttttccca tgtaatatcc ttttctattt ttcgttccgt
     2341 taccaacttt acacatactt tatatagcta ttcacttcta tacactaaaa aactaagaca
     2401 attttaattt tgctgcctgc catatttcaa tttgttataa attcctataa tttatcctat
     2461 tagtagctaa aaaaagatga atgtgaatcg aatcctaaga gaattgtgtt acaaccaatt
     2521 aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     2581 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     2641 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     2701 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     2761 atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg
     2821 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     2881 cattcgtgat tgcgcctgag cgaggcgaaa tacgcgatcg ctgttaaaag gacaattaca
     2941 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     3001 tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag
     3061 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     3121 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     3181 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc
     3241 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     3301 atttaatcgc ggcctggagc aagacgtttc ccgttgaata tggctcataa caccccttgt
     3361 attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc
     3421 aatgtaacat cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc
     3481 tgagttgaag gatcagtcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3541 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3601 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3661 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     3721 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3781 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3841 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3901 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3961 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4021 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4081 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4141 ggggggcgga gcctatggaa aaacgcca
//
