LOCUS       sequence-422259-        4274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid backbone for yeast expression, use auxotrophic marker URA3 
            (Kluyveromyces lactis).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4274)
  AUTHORS   He CW, Cui XF, Ma SJ, Xu Q, Ran YP, Chen WZ, Mu JX, Li H, Zhu J, 
            Gong Q, Xie Z
  TITLE     Membrane recruitment of Atg8 by Hfl1 facilitates turnover of 
            vacuolar membrane proteins in yeast cells approaching stationary 
            phase.
  JOURNAL   BMC Biol. 2021 Jun 4;19(1):117. doi: 10.1186/s12915-021-01048-7.
  PUBMED    34088313
REFERENCE   2  (bases 1 to 4274)
  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: "BMC Biol.";
            date: "2021-06-4"; volume: "19(1):117. doi"; pages: " 
            10.1186/s12915-021-01048-7"
COMMENT     Sequence Label: ClhN-URA3
FEATURES             Location/Qualifiers
     source          1..4274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      101..288
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(156..180)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    340..373
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1825..1858
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(1911..1930)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1912..1930)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2017..2034)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2188..2776)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2268..2287)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2947..3738,3739..3807))
                     /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     3570..3589
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3808..3912)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3980..3998
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4036..4058)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4158..4177
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4258..4274,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4258..4274,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg tgggattcct gggttaatta
       61 acatcgctag ccctaggtga ggcgcgccac ttctaaataa gcgaatttct tatgatttat
      121 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
      181 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
      241 ttctcaggta tagtatgagg tcgctcttat tgaccacacc tctaccggca gatccgctag
      301 ggataacagg gtaatatctg caggtcgaca acccttaata taacttcgta taatgtatgc
      361 tatacgaagt tattaggtct agagatccca atacaacaga tcacgtgatc tttttgtaag
      421 atgaagttga agtagtgttg caccgtgcca atgcaggtgg ctattagatt aaatatgtga
      481 tttgttctat taagtttcct gtataattaa tggggagcgc tgattctctt ttggtacgct
      541 tcccatccag catttctgta tctttcacct tcaaccttag gatctctacc cttggcgaaa
      601 agtcctctgc caacaatgat gatatctgat ccaccactta caacttcgtc gacggttctg
      661 tactgctgac ccaatgcatc gcctttgtcg tctaaaccta cacctggggt catgattagc
      721 caatcaaacc cttcttctct tcctcccata tcgttctgag caatgaaccc aataacgaaa
      781 tctttatcac tctttgcaat atcaacggta cccttagtat attcaccgtg tgctagagaa
      841 cccttggaag acaattcagc aagcatcaat aatccccttg gttctttggt gacctcttgc
      901 gcaccttgtt tcaagccagc aacaatacca gcaccagtaa ccccgtgggc gttggtgata
      961 tcagaccatt ctgcgatacg gtaaacgccc gatgtatatt gtaatttgac tgtgttaccg
     1021 atatcggcga attttctgtc ctcaaatatc aagaacttgt atttctctgc caatgctttc
     1081 aatggaacga cagtaccctc ataactgaaa tcatccaaga tatcaacgtg tgttttcaaa
     1141 aggcaaatgt atggacccaa cgtttcaaca agtttcaata gctcatcagt cgaacgaacg
     1201 tcaagagaag cacacaaatt ggtcttcttt tcatccatta aacgtaaaag tttcgatgca
     1261 accggacttg catgagtctc agctctactg gtatatgatt ttgtggacat ggtgcaacta
     1321 attgacggga gtgtattgac gctggcgtac tggctttcac aaaatggccc aatcacaacc
     1381 acatcttaga tagttgaaat gactttagat aacatcaatt gagatgagct taatcatgtc
     1441 aaagctaaaa gtgtcaccat gaacgacaat tcttaagcaa atcacgtgat atagatccac
     1501 gaataaccac catttgatgc tcgaggcaag taatgtgtgt aaaaaaatgc gttaccacca
     1561 tccaatgcag accgatcttc tacccagaat cacatatatt tatgtaccga gtaccttttt
     1621 tctatcttcc aattgcttct cccatatgat tgtctccgta agctcgaaat ttctaagttg
     1681 gattttaatc ttcacgcagg atgacagttc gatgagcttc tgaggagtgt ttagaacata
     1741 atcatttatc catggtctat ctcttcttgt cgctttttct cctcgataga acctaaataa
     1801 aacgagctct cgagaaccct taatataact tcgtataatg tatgctatac gaagttatta
     1861 ggtgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct ccctatagtg
     1921 agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg
     1981 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2041 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2101 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2161 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2221 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2281 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2341 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2401 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2461 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2521 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2581 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2641 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2701 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2761 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2821 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2881 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2941 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3001 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3061 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3121 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3181 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3241 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3301 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3361 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3421 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3481 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3541 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3601 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3661 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3721 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3781 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     3841 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     3901 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     3961 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4021 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4081 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4141 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatggacat
     4201 attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
     4261 taggtgacac tata
//
