LOCUS       sequence-382506-        6992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CDS1 overexpression. Diverts glycerolipid synthesis pathway towards 
            phospholipids. Uses antibiotic resistance marker hphMX6..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6992)
  AUTHORS   Li D, Yang SG, He CW, Zhang ZT, Liang Y, Li H, Zhu J, Su X, Gong Q, 
            Xie Z
  TITLE     Excess diacylglycerol at the endoplasmic reticulum disrupts 
            endomembrane homeostasis and autophagy.
  JOURNAL   BMC Biol. 2020 Aug 28;18(1):107. doi: 10.1186/s12915-020-00837-w.
  PUBMED    32859196
REFERENCE   2  (bases 1 to 6992)
  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: "2020-08-28"; volume: "18(1):107. doi"; pages: " 
            10.1186/s12915-020-00837-w"
COMMENT     Sequence Label: UG75-CDS1-3HA
FEATURES             Location/Qualifiers
     source          1..6992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1862..1951
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     terminator      1990..2177
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2045..2069)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    2229..2262
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            2317..3892
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        2317..2660
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2661..3689
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             2661..3689
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      3695..3892
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    3955..3988
                     /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(4041..4060)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4042..4060)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4147..4164)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4318..4906)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4398..4417)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5077..5868,5869..5937))
                     /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     5700..5719
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5938..6042)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6110..6128
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6166..6188)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6288..6307
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6388..6406
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6388..6405
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ctgatattta ttgctcgttt ttcgctattt tgatgcagta catctcagaa ttcaacggag
       61 aacgtggata tattaggcac aaagaatcat aaacaaacgc ccatgtaaat aactatcaat
      121 acactacttt ctaaaccaat ctgcaggagc tccaccttga tcgctttggc ttattccagg
      181 cgttttcagg cactgtatat tggtactata tcgctatgta tttgacagtc attatcaacg
      241 gatacccttc ttgaaaattt ttcagatggg atgacatgtg aaaagtgtaa caatcgagaa
      301 ctgtaaacag catgggggag cattcatcaa tattgttatt tagtgtaagc cagattgtga
      361 agacgcactt cttgccaagt atcttcccaa acagcgaacc tgtcatactt aactgttcca
      421 cttccatata cctaaaatgt ctgacaaccc tgagatgaaa ccacatggta cgagcaagga
      481 gattgtggag tcggttactg acgccacctc aaaggcgatt gataaattgc aagaagaact
      541 ccacaaggac gccagcgaat ccgtcacgcc ggtgaccaag gaaagcactg ctgctacaaa
      601 ggaaagcagg aaatacaact ttttcattag aacagtttgg acgtttgtta tgatcagtgg
      661 tttcttcatc accttagcat cgggtcatgc atggtgtata gtgctgattt tgggctgcca
      721 aattgctact tttaaagagt gtattgccgt aacaagtgca tctggtcgcg aaaagaattt
      781 gccattgaca aagacgttga actggtacct tctcttcacc actatctatt acttagatgg
      841 gaaatcactc ttcaagtttt tccaagctac tttttacgag tatcctgtat tgaactttat
      901 cgtaacaaat cataagttta tctgctattg tctctatctt atgggatttg ttctgttcgt
      961 ttgtagttta agaaagggat ttttgaaatt ccagttcgga tcattgtgcg ttactcatat
     1021 ggttcttctt ctggtagtat ttcaggctca tttgatcatc aaaaacgtgc tcaatgggct
     1081 attctggttc ttattaccat gcgggttggt aattgttaat gatatcttcg cctacctgtg
     1141 cggcattaca ttcggtaaga ctaaactaat agaaatctcg cccaagaaaa ctttagaagg
     1201 tttccttggt gcctggtttt tcactgctct agctagtatt atattaacaa ggattctgag
     1261 cccttatact tacttgacgt gccctgttga agatctccat accaattttt tctctaattt
     1321 gacatgtgaa ctaaacccag ttttccttcc acaagtttac agacttccac ccatcttttt
     1381 cgataaagtt caaatcaatt caattacagt aaaaccaatt tatttccatg ctttaaactt
     1441 agctaccttt gcatcattat ttgcgccatt tggaggcttt ttcgcatctg gtctaaagag
     1501 aacttttaaa gttaaagact ttggccactc catcccaggc catggtggta tcacagacag
     1561 ggttgattgc caattcataa tgggttcttt tgccaatttg tactatgaaa cattcatcag
     1621 cgaacacaga ataacagtag atacagtttt atccactatt ttaatgaacc tgaacgacaa
     1681 gcagattata gaattgattg atatcttgat tagattccta tctaaaaaag gtataatatc
     1741 agcgaagaat tttgaaaagt tggctgacat ctttaatgtg accaagaaat cattgaccaa
     1801 tcactctcaa gcttcgtacg ctgcaggtcg tgggattcct gggttaatta acatcgctag
     1861 ctacccatac gatgttcctg actatgcggg ctatccctat gacgtcccgg actatgcagg
     1921 atcctatcca tatgacgttc cagattacgc tgctcagtgc cctaggtgag gcgcgccact
     1981 tctaaataag cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa
     2041 ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg
     2101 agtaactctt tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt
     2161 gaccacacct ctaccggcag atccgctagg gataacaggg taatatctgc aggtcgacaa
     2221 cccttaatat aacttcgtat aatgtatgct atacgaagtt attaggtcta gagatctgtt
     2281 tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc
     2341 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
     2401 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
     2461 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
     2521 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
     2581 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
     2641 acatccgaac ataaacaacc atgggtaaaa agcctgaact caccgcgacg tctgtcgaga
     2701 agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag
     2761 aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct
     2821 gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc
     2881 cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc
     2941 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc
     3001 agccggtcgc ggaggcaatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt
     3061 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg
     3121 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt
     3181 ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc
     3241 acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag
     3301 cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct
     3361 tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc
     3421 atccggagct tgcaggatcg ccacggctcc gggcgtatat gctccgcatt ggtcttgacc
     3481 aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat
     3541 gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa
     3601 gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc
     3661 ccagcactcg tccgagggca aaggaataat cagtactgac aataaaaaga ttcttgtttt
     3721 caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt
     3781 tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc
     3841 gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat
     3901 tcgatactaa cgccgccatc cagtgtcgaa aacgagctct cgagaaccct taatataact
     3961 tcgtataatg tatgctatac gaagttatta ggtgatatca gatccactag tggcctatgc
     4021 ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa gccaggttaa
     4081 cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4141 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4201 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4261 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4321 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4381 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4441 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4501 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4561 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4621 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4681 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4741 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4801 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4861 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4921 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4981 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5041 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5101 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5161 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5221 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5281 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5341 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5401 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5461 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5521 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5581 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5641 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5701 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5761 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5821 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5881 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5941 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6001 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6061 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     6121 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     6181 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     6241 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     6301 gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct acaattaata
     6361 cataacctta tgtatcatac acatacgatt taggtgacac tatagaacgc ggccgccagc
     6421 tgaagcttcc atgatttctt cttctggtgc caacgaatcg aataaggtat cgaaactggt
     6481 tccttttaaa agtcgcttag catccgcatg ccaaaagctt ggcggcaggt ttaagtgccc
     6541 ttcgtcatca cgaattcgga tagttcgcag aaatgggtac caatgactag aatttctgaa
     6601 aactgtagtt tcgtaaatga atgcgatgtt tagagctaac attccatcga tgctactgtc
     6661 acataacaaa ttggcaatgg agctgttaga tgcagaaaaa atcgaggatt tgttcaacgt
     6721 caatattgac tcaccttctg caatatcagt ctttgcaaag acacccaatc caccgagagg
     6781 cgattctctt atatcacatt tggaatcttc aaacctgcca ttgcaattag caacaaactt
     6841 cagaatccga tgaacatcgt ccttaaaagt tacggacatc tcactctttc aaatttattt
     6901 tttaattgct attatatatg gcaacaaatc tttcttattg tatctgccac ctcatctcag
     6961 ctgccttcat cgagtatact tcatttgatt tt
//
