LOCUS       sequence-349767-        4187 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  USER cloning-compatible 2u vector (HIS3).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4187)
  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 4187)
  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: pEDJ437
FEATURES             Location/Qualifiers
     source          1..4187
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        49..264
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             274..936
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1279..1867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1359..1378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2038..2829,2830..2898))
                     /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     2661..2680
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2899..3003)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3030..4012
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(3993..4040)
                     /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 aatcatctgt catatgacag agcagaaagc cctagtaaag
      301 cgtattacaa atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg
      361 atagagcact cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa
      421 tcgcaagtga ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg
      481 gccaagcatt ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac
      541 catcacacca ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctaggg
      601 gccgtgcgtg gagtaaaaag gtttggatca ggatttgcgc ctttggatga ggcactttcc
      661 agagcggtgg tagatctttc gaacaggccg tacgcagttg tcgaacttgg tttgcaaagg
      721 gagaaagtag gagatctctc ttgcgagatg atcccgcatt ttcttgaaag ctttgcagag
      781 gctagcagaa ttaccctcca cgttgattgt ctgcgaggca agaatgatca tcaccgtagt
      841 gagagtgcgt tcaaggctct tgcggttgcc ataagagaag ccacctcgcc caatggtacc
      901 aacgatgttc cctccaccaa aggtgttctt atgtagagtg caggtggaga ttgataagac
      961 ttttctagtt gcatatcttt tatatttaaa tcttatctat tagttaattt tttgtaattt
     1021 atccttatat atagtctggt tattctaaaa tatcatttca gtatctaaaa attcccctct
     1081 tttttcagtt atatcttaac aggcgacagt ccaaatgttg atttatccca gtccgattca
     1141 tcagggttgt gaagcatttt gtcaatggtc gaaatcacat cagtaatagt gcctcttact
     1201 tgcctcatag acatcgacga gttgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1261 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1321 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1381 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1441 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1501 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1561 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1621 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1681 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1741 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1801 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1861 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1921 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1981 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2041 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2101 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2161 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2221 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2281 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2341 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2401 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2461 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2521 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2581 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2641 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2701 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2761 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2821 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2881 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2941 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3001 gcgcacattt ccccgaaaag tgccacctga acgaagcatc tgtgcttcat tttgtagaac
     3061 aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag
     3121 aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt
     3181 aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     3241 tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt
     3301 tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac
     3361 tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt
     3421 ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg
     3481 actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa
     3541 aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt
     3601 gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct
     3661 ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa
     3721 tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt
     3781 agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg
     3841 atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat
     3901 tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc
     3961 ttcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct agagaatagg
     4021 aacttcggaa taggaacttc aaagcgtttc cgaaaacgag cgcttccgaa aatgcaacgc
     4081 gagctgcgca catacagctc actgttcacg ttgcacctat atctgcgtgt tgcctgtata
     4141 tatatataca tgagaagaac ggcatagtgc gtgtttatgc ttaaatg
//
