LOCUS       sequence-471571-        5665 bp    DNA     circular SYN 05-MAY-2025
DEFINITION  2µ plasmid for constitutive expression of TP901 under RNR2 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5665)
  AUTHORS   Pisera A, Yu Y, Williams RL, Liu CC
  TITLE     Ultra-Efficient Integration of Gene Libraries onto Yeast Cytosolic 
            Plasmids.
  JOURNAL   bioRxiv [Preprint]. 2024 Nov 30:2024.11.29.626108. doi: 
            10.1101/2024.11.29.626108.
  PUBMED    39651169
REFERENCE   2  (bases 1 to 5665)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Nov 30:2024.11.29.626108. doi: 
            10.1101/2024.11.29.626108."
COMMENT     Sequence Label: pYY10-2µ-TP901
FEATURES             Location/Qualifiers
     source          1..5665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      471..668
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     terminator      2892..3081
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(3138..3726)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3218..3237)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      3748..4730
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(4711..4758)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     gene            join(4977..5665,1..668)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        4977..5320
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             join(5321..5665,1..465)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5388..5407)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
       61 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
      121 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
      181 gtctggaaag aaatgcataa gcttttgcca ttctcaccgg attcagtcgt cactcatggt
      241 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
      301 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
      361 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
      421 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcagt actgacaata
      481 aaaagattct tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc
      541 tattttaatc aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag
      601 atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg
      661 ctatactgct gtcgattcga tactaacgcc gccatccagt gtcgaaaacg agctctcgag
      721 aacccttaat agtcgaacaa gaagcaggca aagtttagag cactgcccct ccgcactcaa
      781 aaaagaaaaa actaggagga aaataaaatt ctcaaccaca caaacacata aacacataca
      841 aatacaaata caagcttatt tacttgacat cgcgcgatct tccactattc agcgccgtcc
      901 gccctctctc gtgttttttg tttacgcgac aactatgcga aatccggagc aacgggcaac
      961 cgtttgggga aagaccacac ccacgcgcga tcgccatggc aacgaggtcg cacacgcccc
     1021 acacccagac ctccctgcga gcgggcatgg gtacaatgtc cccgttgcca cagacaccac
     1081 ttcgtagcac agcgcagagc gtagcgtgtt gttgctgctg acaaaagaaa atttttctta
     1141 gcaaagcaaa ggaggggaag cacgggcaga tagcaccgta ccataccctt ggaaactcga
     1201 aatgaacgaa gcaggaaatg agagaatgag agttttgtag gtatatatag cggtagtgtt
     1261 tgcgcgttac catcatcttc tggatctatc tattgttctt ttcctcatca ctttcccctt
     1321 tttcgctctt cttcttgtct tttatttctt tctttttttt aattgttccc tcgattggct
     1381 atctaccaaa gaatccaaac ttaatacacg tatttatttg tccaattacc atgacgaaaa
     1441 aggtcgcaat ctatactcgt gtttccacaa caaatcaagc cgaagaagga tttagcatag
     1501 atgaacaaat cgaccgtctg acaaaatatg ctgaggcgat ggggtggcag gttagcgaca
     1561 cgtacactga cgcaggattc tctggagcaa aactagaaag acctgcaatg cagcgtctta
     1621 ttaatgacat agaaaacaag gcgttcgata cagttctagt ctacaaattg gataggctat
     1681 ctcgttctgt gagagacaca ctatacctag ttaaagatgt ctttaccaaa aacaagatag
     1741 acttcatatc tctgaacgaa agcatagata catccagcgc aatgggatca ttgtttctta
     1801 caattttatc agctataaac gaatttgaaa gagaaaacat caaggaaagg atgacgatgg
     1861 gcaagctggg cagagccaaa agcgggaagt caatgatgtg gacaaagacc gctttcggct
     1921 actaccacaa cagaaagacg gggatcttgg agatagttcc tttgcaggct accattgtag
     1981 aacagatttt tacggattac ctttccggga tctccctgac aaaacttcgt gataaattga
     2041 acgaatcagg ccatatcggc aaagatatac cctggagcta tagaactctt cgtcagacgc
     2101 tggataatcc ggtttattgt gggtatataa aattcaaaga cagtttgttc gaaggtatgc
     2161 acaagcccat cattccatat gaaacgtatc tgaaagtgca gaaagagctg gaagaaaggc
     2221 agcagcagac ctacgaaagg aataacaatc ctagaccctt ccaagcgaag tatatgttga
     2281 gcgggatggc taggtgtggg tactgcggtg ctcctctgaa aatagtattg gggcataaga
     2341 ggaaagacgg cagtaggaca atgaaatatc attgtgccaa taggttcccg cgtaaaacaa
     2401 aagggattac cgtttataat gataataaga agtgcgacag cggcacatat gatttaagca
     2461 acctggaaaa tactgtgatc gacaacctga tcggtttcca agagaataat gattcccttt
     2521 taaaaatcat caatggtaat aaccagccaa tactagacac aagctcattc aagaagcaaa
     2581 tttcccaaat agacaagaaa atccaaaaaa atagcgacct atacttaaat gactttatca
     2641 caatggatga gttaaaagac agaactgata gcttacaggc agaaaagaaa ttactgaagg
     2701 cgaagatatc agagaacaag ttcaacgact caactgacgt attcgaacta gtaaaaaccc
     2761 aactgggcag cataccaata aatgaactgt catacgacaa caagaagaag atcgtgaaca
     2821 atcttgtaag taaggtggac gtgacagcag ataatgttga tataatcttc aagttccagt
     2881 tagcataata aatccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
     2941 cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
     3001 cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag
     3061 aaggttttgg gacgctcgaa gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3121 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3181 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3241 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3301 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3361 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3421 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3481 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3541 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3601 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3661 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3721 tctcaagaag atcctttgat cttttctacc gaagcatctg tgcttcattt tgtagaacaa
     3781 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     3841 cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa
     3901 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     3961 cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt
     4021 tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt
     4081 tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc
     4141 gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac
     4201 tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag
     4261 gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga
     4321 tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct
     4381 atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata
     4441 gttcttacta caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag
     4501 aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat
     4561 atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc
     4621 gcaatatttt agtagcccgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtctt
     4681 cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag agaataggaa
     4741 cttcggaata ggaacttcaa agcgtttccg aaaacgagcg cttccgaaaa tgcaacgcga
     4801 gctgcgcaca tacagctcac tgttcacgtc gcacctatat ctgcgtgttg cctgtatata
     4861 tatatacatg agaagaacgg catagtgcgt gtttatgctt aaatgcgtat atgtgttatg
     4921 tataggtcta gagatctgtt tagcttgcct cgtccccgcc gggtcacccg gccagcgaca
     4981 tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg
     5041 actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca tccatacatt
     5101 ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcagacc tgcgagcagg
     5161 gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg tagagaaata
     5221 taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt aaaatcttgc
     5281 taggatacag ttctcacatc acatccgaac ataaacaacc atgggtaagg aaaagactca
     5341 cgtttcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     5401 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc
     5461 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     5521 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     5581 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggcaaaacag cattccaggt
     5641 attagaagaa tatcctgatt caggt
//
