LOCUS       sequence-471367-        3524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmids for generation of donor DNA for construction of a 
            BXB1-compatible landing pad p1 with URA3 selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3524)
  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 3524)
  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: pRW398-URA3-BXB1-LP
FEATURES             Location/Qualifiers
     source          1..3524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             300..1103
                     /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"
     protein_bind    1104..1141
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     primer_bind     1360..1382
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1420..1438)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1506..1610
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1611..1679,1680..2471)
                     /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(1829..1848)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2642..3230
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3131..3150
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3384..3401
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 actatctatt ttttcagtac ataatttaat taaatcatca tgacctttat ctaattttac
       61 atctcttcta ccaagaccta attctttatt aatactatca aaatctgtta ttctattgcc
      121 ttgaggatct ttgaaacacc aatctgagtc tgagggcatg cggcttgtcg acgacggcgt
      181 gctccgtcgt caggatcatt ttttcgccgg agtcaattag gtcatacttt tctatataat
      241 ccaaatccca aaaatcaatt gaatgattct taatatgatt taatagttta tgattataaa
      301 tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc
      361 tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca
      421 ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac
      481 atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat
      541 ccgccaagta caatttttta ctcttcgagg acagaaaatt tgctgacatt ggtaatacag
      601 tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg
      661 cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa
      721 caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta
      781 ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg
      841 gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga
      901 cacccggtgt gggtttagat gacaagggag atgcattggg tcaacagtat agaaccgtgg
      961 atgatgtggt ttctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg
     1021 gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga
     1081 gaagatgcgg ccagcaaaac taaatgatcc tgacgacgga gagcgccgtc gtcgacaagc
     1141 caatccataa tggcttcatc taatgtggac tcttctgagc cccgtatttc cagagaaagc
     1201 tctttgaaga ggtctctatc tatttcaaag tcgttaaaag ggttgttcaa gtcaggaggg
     1261 ggaaacgcca atactggccc tactactgct gctgctgctg ctgcaccatc tagtatcttt
     1321 taaattaata atgtaattca tatattatag tactcgtctc cgggagctgc atgtgtcaga
     1381 ggttttcacc gtcatcaccg aaacgcgtga gacgaaaggg cctcgtgata cgcctatttt
     1441 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     1501 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1561 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1621 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1681 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1741 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1801 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     1861 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1921 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     1981 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2041 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2101 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     2161 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2221 aattaataga ctggatggag gtggataaag ttgcaggacc acttctgcgc tcggcccttc
     2281 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2341 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2401 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2461 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     2521 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2581 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2641 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2701 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2761 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     2821 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2881 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2941 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3001 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3061 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3121 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3181 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3241 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3301 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3361 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     3421 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     3481 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tagt
//
