LOCUS       sequence-392931-        6710 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 6710)
  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 6710)
  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-ADH1-CDS1-3HA
FEATURES             Location/Qualifiers
     source          1..6710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        53..71
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     53..70
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    122..155
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            210..1785
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        210..553
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             554..1582
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             554..1582
                     /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      1588..1785
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    1848..1881
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1922..2626
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             4082..4171
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     terminator      4210..4397
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4265..4289)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(4416..4435)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4417..4435)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4522..4539)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4693..5281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4773..4792)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5452..6243,6244..6312))
                     /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     6075..6094
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6313..6417)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6485..6503
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6541..6563)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6663..6682
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgttagaacg cggctacaat taatacataa ccttatgtat catacacata cgatttaggt
       61 gacactatag aacgcggccg ccagctgaag cttcgtacgc tgcaggtcga caacccttaa
      121 tataacttcg tataatgtat gctatacgaa gttattaggt ctagagatct gtttagcttg
      181 cctcgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc ctccttgaca
      241 gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt tagcccatac
      301 atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg cgaagcaaaa
      361 attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca gacgcgttga
      421 attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg ccactgaggt
      481 tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac atcacatccg
      541 aacataaaca accatgggta aaaagcctga actcaccgcg acgtctgtcg agaagtttct
      601 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
      661 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
      721 tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc
      781 ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc
      841 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
      901 cgcggaggca atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc
      961 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     1021 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     1081 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     1141 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     1201 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     1261 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
     1321 gcttgcagga tcgccacggc tccgggcgta tatgctccgc attggtcttg accaactcta
     1381 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     1441 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc
     1501 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
     1561 tcgtccgagg gcaaaggaat aatcagtact gacaataaaa agattcttgt tttcaagaac
     1621 ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg
     1681 atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa
     1741 gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc gattcgatac
     1801 taacgccgcc atccagtgtc gaaaacgagc tctcgagaac ccttaatata acttcgtata
     1861 atgtatgcta tacgaagtta ttaggtgata tcagatccac tagtggccta tgcggccgcg
     1921 gatccttttg ttgtttccgg gtgtacaata tggacttcct cttttctggc aaccaaaccc
     1981 atacatcggg attcctataa taccttcgtt ggtctcccta acatgtaggt ggcggagggg
     2041 agatatacaa tagaacagat accagacaag acataatggg ctaaacaaga ctacaccaat
     2101 tacactgcct cattgatggt ggtacataac gaactaatac tgtagcccta gacttgatag
     2161 ccatcatcat atcgaagttt cactaccctt tttccatttg ccatctattg aagtaataat
     2221 aggcgcatgc aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc
     2281 atatccgcaa tgacaaaaaa atgatggaag acactaaagg aaaaaattaa cgacaaagac
     2341 agcaccaaca gatgtcgttg ttccagagct gatgaggggt atctcgaagc acacgaaact
     2401 ttttccttcc ttcattcacg cacactactc tctaatgagc aacggtatac ggccttcctt
     2461 ccagttactt gaatttgaaa taaaaaaaag tttgctgtct tgctatcaag tataaataga
     2521 cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt
     2581 ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactatct catatacaat
     2641 gtctatccca atcgatatgt ctgacaaccc tgagatgaaa ccacatggta cgagcaagga
     2701 gattgtggag tcggttactg acgccacctc aaaggcgatt gataaattgc aagaagaact
     2761 ccacaaggac gccagcgaat ccgtcacgcc ggtgaccaag gaaagcactg ctgctacaaa
     2821 ggaaagcagg aaatacaact ttttcattag aacagtttgg acgtttgtta tgatcagtgg
     2881 tttcttcatc accttagcat cgggtcatgc atggtgtata gtgctgattt tgggctgcca
     2941 aattgctact tttaaagagt gtattgccgt aacaagtgca tctggtcgcg aaaagaattt
     3001 gccattgaca aagacgttga actggtacct tctcttcacc actatctatt acttagatgg
     3061 gaaatcactc ttcaagtttt tccaagctac tttttacgag tatcctgtat tgaactttat
     3121 cgtaacaaat cataagttta tctgctattg tctctatctt atgggatttg ttctgttcgt
     3181 ttgtagttta agaaagggat ttttgaaatt ccagttcgga tcattgtgcg ttactcatat
     3241 ggttcttctt ctggtagtat ttcaggctca tttgatcatc aaaaacgtgc tcaatgggct
     3301 attctggttc ttattaccat gcgggttggt aattgttaat gatatcttcg cctacctgtg
     3361 cggcattaca ttcggtaaga ctaaactaat agaaatctcg cccaagaaaa ctttagaagg
     3421 tttccttggt gcctggtttt tcactgctct agctagtatt atattaacaa ggattctgag
     3481 cccttatact tacttgacgt gccctgttga agatctccat accaattttt tctctaattt
     3541 gacatgtgaa ctaaacccag ttttccttcc acaagtttac agacttccac ccatcttttt
     3601 cgataaagtt caaatcaatt caattacagt aaaaccaatt tatttccatg ctttaaactt
     3661 agctaccttt gcatcattat ttgcgccatt tggaggcttt ttcgcatctg gtctaaagag
     3721 aacttttaaa gttaaagact ttggccactc catcccaggc catggtggta tcacagacag
     3781 ggttgattgc caattcataa tgggttcttt tgccaatttg tactatgaaa cattcatcag
     3841 cgaacacaga ataacagtag atacagtttt atccactatt ttaatgaacc tgaacgacaa
     3901 gcagattata gaattgattg atatcttgat tagattccta tctaaaaaag gtataatatc
     3961 agcgaagaat tttgaaaagt tggctgacat ctttaatgtg accaagaaat cattgaccaa
     4021 tcactctcaa gcttcgtacg ctgcaggtcg tgggattcct gggttaatta acatcgctag
     4081 ctacccatac gatgttcctg actatgcggg ctatccctat gacgtcccgg actatgcagg
     4141 atcctatcca tatgacgttc cagattacgc tgctcagtgc cctaggtgag gcgcgccact
     4201 tctaaataag cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa
     4261 ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg
     4321 agtaactctt tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt
     4381 gaccacacct ctaccggccg cggatctgcc ggtctcccta tagtgagtcg tattaatttc
     4441 gataagccag gttaacctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4501 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4561 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4621 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4681 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4741 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4801 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4861 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4921 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4981 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5041 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5101 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5161 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5221 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5281 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5341 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5401 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5461 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5521 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5581 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5641 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5701 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5761 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5821 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5881 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5941 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6001 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6061 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6121 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6181 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6241 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6301 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6361 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6421 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     6481 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     6541 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     6601 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     6661 tgcggcatca gagcagattg tactgagagt gcaccatatg gacatattgt
//
