LOCUS       sequence-471411-        3518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids for generation of donor DNA for construction of a 
            TP901-compatible landing pad p1 with URA3 selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3518)
  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 3518)
  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]. 2024 Nov 30:2024.11.29.626108. doi: 
            10.1101/2024.11.29.626108."
COMMENT     Sequence Label: pYY13-URA3-TP901LP
FEATURES             Location/Qualifiers
     source          1..3518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      686..1274
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1175..1194
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1428..1445
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             1865..2668
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     primer_bind     2922..2944
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2982..3000)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3068..3172
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3173..3241,3242..3518,1..515)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEVDKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3391..3410)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
       61 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
      121 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
      181 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
      241 tacttactct agcttcccgg caacaattaa tagactggat ggaggtggat aaagttgcag
      301 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
      361 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
      421 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
      481 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
      541 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
      601 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
      661 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
      721 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      781 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
      841 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      901 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      961 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1021 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     1081 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     1141 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     1201 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     1261 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     1321 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     1381 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     1441 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     1501 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     1561 ttaatagtac tatctatttt ttcagtacat aatttaatta aatcatcatg acctttatct
     1621 aattttacat ctcttctacc aagacctaat tctttattaa tactatcaaa atctgttatt
     1681 ctattgcctt gaggatcttt gaaacaccaa tctgagtctg agggcatgca acacaattaa
     1741 catctcaatc aaggtaaatg cttttttttc gccggagtca attaggtcat acttttctat
     1801 ataatccaaa tcccaaaaat caattgaatg attcttaata tgatttaata gtttatgatt
     1861 ataaatgtcg aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc
     1921 caagctattt aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg
     1981 taccaccaag gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa
     2041 aacacatgtg gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc
     2101 attatccgcc aagtacaatt ttttactctt cgaggacaga aaatttgctg acattggtaa
     2161 tacagtcaaa ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac
     2221 gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga
     2281 agtaacaaag gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct
     2341 atctactgga gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt
     2401 tatcggcttt attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat
     2461 tatgacaccc ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac agtatagaac
     2521 cgtggatgat gtggtttcta caggatctga cattattatt gttggaagag gactatttgc
     2581 aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata
     2641 tttgagaaga tgcggccagc aaaactaaaa agcatttacc ttgatttgga tgttaattgt
     2701 gttaatccat aatggcttca tctaatgtgg actcttctga gccccgtatt tccagagaaa
     2761 gctctttgaa gaggtctcta tctatttcaa agtcgttaaa agggttgttc aagtcaggag
     2821 ggggaaacgc caatactggc cctactactg ctgctgctgc tgctgcacca tctagtatct
     2881 tttaaattaa taatgtaatt catatattat agtactcgtc tccgggagct gcatgtgtca
     2941 gaggttttca ccgtcatcac cgaaacgcgt gagacgaaag ggcctcgtga tacgcctatt
     3001 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3061 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3121 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3181 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3241 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3301 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3361 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3421 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3481 ctcaccagtc acagaaaagc atcttacgga tggcatga
//
