LOCUS       sequence-472629-        8179 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression of ScVPS34 HELCAT .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8179)
  AUTHORS   Rostislavleva K, Soler N, Ohashi Y, Zhang L, Pardon E, Burke JE, 
            Masson GR, Johnson C, Steyaert J, Ktistakis NT, Williams RL
  TITLE     Structure and flexibility of the endosomal Vps34 complex reveals the
            basis of its function on membranes.
  JOURNAL   Science. 2015 Oct 9;350(6257):aac7365. doi: 10.1126/science.aac7365.
  PUBMED    26450213
REFERENCE   2  (bases 1 to 8179)
  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: "Science."; 
            date: "2015-10-9"; volume: "350(6257):aac7365. doi"; pages: " 
            10.1126/science.aac7365"
COMMENT     Sequence Label: pYO361
FEATURES             Location/Qualifiers
     source          1..8179
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        191..472
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             473..1147
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1250..1705)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1387..1408)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1599..1618
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1831..1853
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1845..1862
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1846..1862
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1872..1891
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1872..1890
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4171..4194)
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     protein_bind    4375..4492
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(4577..4597)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4577..4595)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4616..4632)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4616..4632)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4629..4651)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4640..4656
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4664..4670,4671..4688,4689..4694))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4709..4730
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4847..4864)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5018..5606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5098..5117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5777..6568,6569..6637))
                     /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     6400..6419
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6638..6742)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6769..8111
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(7732..7779)
                     /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)."
     primer_bind     8152..8170
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatagatc aacgacatta ctatatatat aatataggaa gcatttaata gaacagcatc
      241 gtaatatatg tgtactttgc agttatgacg ccagatggca gtagtggaag atattcttta
      301 ttgaaaaata gcttgtcacc ttacgtacaa tcttgatccg gagcttttct ttttttgccg
      361 attaagaatt aattcggtcg aaaaaagaaa aggagagggc caagagggag ggcattggtg
      421 actattgagc acgtgagtat acgtgattaa gcacacaaag gcagcttgga gtatgtctgt
      481 tattaatttc acaggtagtt ctggtccatt ggtgaaagtt tgcggcttgc agagcacaga
      541 ggccgcagaa tgtgctctag attccgatgc tgacttgctg ggtattatat gtgtgcccaa
      601 tagaaagaga acaattgacc cggttattgc aaggaaaatt tcaagtcttg taaaagcata
      661 taaaaatagt tcaggcactc cgaaatactt ggttggcgtg tttcgtaatc aacctaagga
      721 ggatgttttg gctctggtca atgattacgg cattgatatc gtccaactgc atggagatga
      781 gtcgtggcaa gaataccaag agttcctcgg tttgccagtt attaaaagac tcgtatttcc
      841 aaaagactgc aacatactac tcagtgcagc ttcacagaaa cctcattcgt ttattccctt
      901 gtttgattca gaagcaggtg ggacaggtga acttttggat tggaactcga tttctgactg
      961 ggttggaagg caagagagcc ccgaaagctt acattttatg ttagctggtg gactgacgcc
     1021 agaaaatgtt ggtgatgcgc ttagattaaa tggcgttatt ggtgttgatg taagcggagg
     1081 tgtggagaca aatggtgtaa aagactctaa caaaatagca aatttcgtca aaaatgctaa
     1141 gaaataggtt attactgagt agtatttatt taagtattgt ttgtgcactt gccgatctat
     1201 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc
     1261 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     1321 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     1381 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     1441 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     1501 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     1561 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     1621 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     1681 cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag
     1741 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     1801 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     1861 gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccgggccc cccctcgagc
     1921 ggtccggcgc gccggccgcc agatcgatct tcgaggggaa ttctcttagg attcgattca
     1981 cattcatctt tttttagcta ctaataggat aaattatagg aatttataac aaattgaaat
     2041 atggcaggca gcaaaattaa aattgtctta gttttttagt gtatagaagt gaatagctat
     2101 ataaagtatg tgtaaagttg gtaacggaac gaaaaataga aaaggatatt acatgggaaa
     2161 acatgttgtt tacggagaaa tgaaaagtat attgtatttt gtacgagcta aaagtacagt
     2221 gggaacaaag tcgacttagg tccgccagta ttgtgccaga ttatgtaaat gatcaatcac
     2281 gataggcagc aaagcattta cactatcatt gattagattc tgaaaatgca ctgtggcatc
     2341 ttcttcggac atattcaaat taaatctctc tcttacacgt aatatagcac cattgggatc
     2401 tattctgata tcaggtatgt tcgacgtctt cattaattca aacaggttta agattaagcc
     2461 tgcattcctt cttaaaatcg aatatgccac aaaacagtag ctgcgaaatt tatcataatt
     2521 tgatgattct gcacccccaa acgcctctat aatttgaggg ggcaatttca ttaatggcgg
     2581 aaaaggtttg gggtcctgac ccaagatata accaaagtct gcgtgaaaaa agtgcccatc
     2641 tggtgtgact agtaagttgt ctaagtgcct atcgccgaca cctaagatgt atgtaataac
     2701 acaatagcca gcacaagatt tgacaaaatt atctaaaacc cagccctgta cgcctaatgt
     2761 cgcgttctca tcaggatagt ggagtttaag gtagccaaga ataccgtgat atttgcttag
     2821 tatgctagct aatgtgtcat tagggataaa ttcaatggca ccttcttgtg gtcctgttgc
     2881 caaaattttg tatggtgtca atttcaagtc cacgttttcg ttttttaata actcattcat
     2941 taaacttatg atctgtacta ccagttgatc ttgtctcaaa tcatcgccaa ccttaaacat
     3001 taagtgatat ggttggttta aggtcgtttt gaatgttatt tttagtggcg ataaggagct
     3061 tttaaatacc ttcgaagttt caggacaaac gtcgcatatc aacacatcag ggtctagtgg
     3121 taaagcaatt ggtcgtactt tgacaagagg cctgactttc gtctccaata aatgcactag
     3181 taattccatc tttttggctg tagtatcttt cagcctctta attgtttcac aacattccct
     3241 gagcacattt attagcctga cttgatcgtt cagtatattc cgagattttt tatccagtct
     3301 actccaaaag gaacttaaaa tctggtctaa atatggcttg tcctcagact cggactttaa
     3361 ataccaatag aagaaacttc ctaaccttgg gttgaccagt gctctcctaa tcaaaaactc
     3421 agccaaggga gaaatgacga ttggtagcga ttctgtcccg gaagtttccg attcactcga
     3481 gatggacttt aataattttt ggttggcatg cgatgtagat agtaacatag aatcacccga
     3541 aagcttttgc gaagatacgg catccacaat agtgaattca ctgttagatt tgtcggaaaa
     3601 agtggacagg ttctcaaaac agacagcttc taccaactgt aataagtaca attccagctc
     3661 cttgtctgat gccttcttca gcctatttac ggcgtacgat ctcactgaaa ggtttttaaa
     3721 agttgagcct aatagttcta atgcgtcgtc tatatcaatt tctgcccatg agtccattag
     3781 ttctaaaacc tctactcttt ccgattcttc tctcaaattt gtactctgta ataactttgt
     3841 aagagctttt ttattgttca tcaaataata cctatatttc catattgaac ccttttcatg
     3901 tgctgttaat ttagtgccag gggggtagtt gatgatttta ttcaagtagt ccctcttttt
     3961 aatgtccggc ttcacttgct tgtccaagtt tgcattttta gatgctcttt ccaatctcct
     4021 atacttttcc tctattggat cattgtttgg ttggtcagga tccatggttt atttatgtgt
     4081 gtttattcga aactaagttc ttggtgtttt aaaactaaaa aaaagactaa ctataaaagt
     4141 agaattcaag aagtttaaga aatagattta cagaattaca atcaatacct accgtcttta
     4201 tatacttatt agtcaagtag gggaataatt tcagggaact ggtttaaacc ttttttttca
     4261 gtcgagatat ctggttgtta atttgattcg ttcatttgag gtttgtgggg ccaggttact
     4321 gccaattttc ctcttcataa ccataaaagc tagtattgta gaatctttat tgttcggagc
     4381 agtgcggcgc gaggcacatc tgcgtttcag gaacgcgacc ggtgaagacg aggacgcacg
     4441 gaggagagtc ttccttcgga gggctgtcac ccgctcggcg gcttctaatc cgtacgcatg
     4501 ctttttttgg atctttgcaa agatccgcgg ccggcgcgcc gcggccgcca ccgcggtgga
     4561 gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     4621 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4681 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4741 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4801 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4861 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4921 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4981 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5041 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5101 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5161 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5221 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5281 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5341 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5401 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5461 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5521 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5581 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5641 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5701 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5761 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5821 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5881 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5941 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6001 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6061 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6121 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6181 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6241 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6301 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6361 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6421 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6481 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6541 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6601 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6661 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6721 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgaa cgaagcatct
     6781 gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     6841 ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga
     6901 atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     6961 agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac
     7021 aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta
     7081 acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat
     7141 aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct
     7201 cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg
     7261 gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca
     7321 tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac
     7381 ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt
     7441 tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag
     7501 agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg
     7561 atgggtaggt tatataggga tatagcacag agatatatag caaagagata cttttgagca
     7621 atgtttgtgg aagcggtatt cgcaatattt tagtagctcg ttacagtccg gtgcgttttt
     7681 ggttttttga aagtgcgtct tcagagcgct tttggttttc aaaagcgctc tgaagttcct
     7741 atactttcta gagaatagga acttcggaat aggaacttca aagcgtttcc gaaaacgagc
     7801 gcttccgaaa atgcaacgcg agctgcgcac atacagctca ctgttcacgt cgcacctata
     7861 tctgcgtgtt gcctgtatat atatatacat gagaagaacg gcatagtgcg tgtttatgct
     7921 taaatgcgta cttatatgcg tctatttatg taggatgaaa ggtagtctag tacctcctgt
     7981 gatattatcc cattccatgc ggggtatcgt atgcttcctt cagcactacc ctttagctgt
     8041 tctatatgct gccactcctc aattggatta gtctcatcct tcaatgctat catttccttt
     8101 gatattggat catactaaga aaccattatt atcatgacat taacctataa aaataggcgt
     8161 atcacgaggc cctttcgtc
//
