LOCUS       sequence-349766-        4319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  USER cloning-compatible 2u vector (URA3).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4319)
  AUTHORS   Jensen ED, Laloux M, Lehka BJ, Pedersen LE, Jakociunas T, Jensen MK,
            Keasling JD
  TITLE     A synthetic RNA-mediated evolution system in yeast.
  JOURNAL   Nucleic Acids Res. 2021 Sep 7;49(15):e88. doi: 10.1093/nar/gkab472.
  PUBMED    34107026
REFERENCE   2  (bases 1 to 4319)
  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: "Nucleic 
            Acids Res."; date: "2021-09-7"; volume: "49(15):e88. doi"; pages: " 
            10.1093/nar/gkab472"
COMMENT     Sequence Label: pEDJ400
FEATURES             Location/Qualifiers
     source          1..4319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        49..264
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             265..1068
                     /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
                     ELVEXLGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1411..1999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1491..1510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2170..2961,2962..3030))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2793..2812
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3031..3135)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3162..4144
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(4125..4172)
                     /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)."
ORIGIN
        1 acctgcactc aacggaatgc gtgcgatcgc gtgcattcca taaagctttt caattcatca
       61 tttttttttt attctttttt ttgatttcgg tttccttgaa atttttttga ttcggtaatc
      121 tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata
      181 tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa
      241 acctgcagga aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact
      301 catcctagtc ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg
      361 tgtgcttcat tggatgttcg taccaccaag gaattactgg agttagttga agyattaggt
      421 cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc
      481 acagttaagc cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaagacaga
      541 aaatttgctg acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata
      601 gcagaatggg cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt
      661 ttgaagcagg cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa
      721 ttgtcatgca agggctccct atctactgga gaatatacta agggtactgt tgacattgcg
      781 aagagcgaca aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat
      841 gaaggttacg attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagacgca
      901 ttgggtcaac agtatagaac cgtggatgat gtggtctcta caggatctga cattattatt
      961 gttggaagag gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga
     1021 aaagcaggct gggaagcata tttgagaaga tgcggccagc aaaactaaat ctgtcatgga
     1081 gattgataag acttttctag ttgcatatct tttatattta aatcttatct attagttaat
     1141 tttttgtaat ttatccttat atatagtctg gttattctaa aatatcattt cagtatctaa
     1201 aaattcccct cttttttcag ttatatctta acaggcgaca gtccaaatgt tgatttatcc
     1261 cagtccgatt catcagggtt gtgaagcatt ttgtcaatgg tcgaaatcac atcagtaata
     1321 gtgcctctta cttgcctcat agacatcgac gagttgagca aaaggccagc aaaaggccag
     1381 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1441 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1501 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1561 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1621 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1681 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1741 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1801 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1861 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1921 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1981 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2041 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2101 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2161 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2221 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2281 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     2341 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2401 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2461 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2521 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2581 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2641 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2701 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     2761 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     2821 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     2881 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     2941 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3001 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3061 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3121 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gaacraagca tctgtgcttc
     3181 attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     3241 gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg
     3301 cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     3361 gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat
     3421 ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc
     3481 atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt
     3541 tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat
     3601 aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt
     3661 ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg
     3721 tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct
     3781 tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat
     3841 tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa
     3901 cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta
     3961 ggttatatag ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg
     4021 tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt
     4081 tgaaagtgcg tcttcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt
     4141 ctagagaata ggaacttcgg aataggaact tcaaagcgtt tccgaaaacg agcgcttccg
     4201 aaaatgcaac gcgagctgcg cacatacagc tcactgttca cgtygcacct atatctgcgt
     4261 gttgcctgta tatatatata catgagaaga acggcatagt gcgtgtttat gcttaaatg
//
