LOCUS       sequence-472468-        7805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ScVPS34 delta C154-V202-3xFLAG under its own promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7805)
  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 7805)
  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: pYO933
FEATURES             Location/Qualifiers
     source          1..7805
                     /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        194..409
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             410..1213
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1344..1799)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1481..1502)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1693..1712
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1925..1947
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1939..1956
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1940..1956
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1966..1984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2187..2198
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             4694..4759
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      4776..4963
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4831..4855)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(5032..5052)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5032..5050)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5071..5087)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5071..5087)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5084..5106)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5095..5111
                     /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(5119..5125,5126..5143,5144..5149))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5164..5185
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5302..5319)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5473..6061)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5553..5572)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6232..7023,7024..7092))
                     /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     6855..6874
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7093..7197)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7234..7737
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7778..7796
                     /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 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
      241 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
      301 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
      361 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc
      421 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
      481 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
      541 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
      601 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
      661 caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca
      721 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
      781 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
      841 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
      901 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
      961 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     1021 gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt
     1081 ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     1141 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     1201 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     1261 agcttcaatt taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg
     1321 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     1381 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1441 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     1501 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1561 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     1621 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     1681 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     1741 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     1801 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1861 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1921 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta
     1981 tagggcgaat tgggtaccgg ggctgaagct ttcgacaaat cttaaattgt gactttgtgc
     2041 atgaaatttt tatatgaacg ttttaaataa gtggacatag tccttcattc ggtatatgca
     2101 gatccggcat caaacacttt ttcaagaaag tttgagtcaa aaggattaaa ggataaagta
     2161 aaacaacaat aacatctccg tgaagcattg agggaagggt ttaactccaa caatgtcact
     2221 gaacaacata acattctgtg tctcacagga tctggatgtt cccctgaaag tgaaaatcaa
     2281 gtcattggaa ggacataagc cactgttgaa gccatctcaa aaaatcctga accctgaatt
     2341 aatgctgata gggtcaaatg tattcccttc tagtgatcta attgtatcct tacaggtatt
     2401 tgataaagag agaaacagaa atttaactct tccgatatac acgccgtata ttcctttcag
     2461 aaactctaga acctgggatt attggttgac tttgcccata cgtatcaaac agcttacctt
     2521 tagcagtcat ttacgcatta ttttgtggga atacaatgga tccaagcaaa ttcccttttt
     2581 caatctagaa acgagcatct ttaacttaaa agactgtact ttaaaaagag ggtttgaatc
     2641 cttaaaattt cgttatgatg tgattgacca cttaaactta gaatttccaa tgttagagct
     2701 tcctgttgtg ttcatcgaaa gggaaattat gaatactcaa atgaatattc caactttgaa
     2761 gaacaatcct ggactaagca cagacttacg agaaccgaat aggaatgatc ctcaaatcaa
     2821 aatttctttg ggggacaaat atcactccac attgaagttt tacgatcctg accaaccaaa
     2881 caatgatcca atagaggaaa agtataggag attggaaaga gcatctaaaa atgcaaactt
     2941 ggacaagcaa gtgaagccgg acattaaaaa gagggactac ttgaataaaa tcatcaacta
     3001 cccccctggc actaaattaa cagcacatga aaagggttca atatggaaat ataggtatta
     3061 tttgatgaac aataaaaaag ctcttacaaa gttattacag agtacaaatt tgagagaaga
     3121 atcggaaaga gtagaggttt tagaactaat ggactcatgg gcagaaattg atatagacga
     3181 cgcattagaa ctattaggct caacttttaa aaacctttca gtgagatcgt acgccgtaaa
     3241 taggctgaag aaggcatcag acaaggagct ggaattgtac ttattacagt tggtagaagc
     3301 tgtctgtttt gagaacctgt ccactttttc cgacaaatct aacagtgaat tcactattgt
     3361 ggatgccgta tcttcgcaaa agctttcggg tgattctatg ttactatcta catcgcatgc
     3421 caaccaaaaa ttattaaagt ccatctcgag tgaatcggaa acttccggga cagaatcgct
     3481 accaatcgtc atttctccct tggctgagtt tttgattagg agagcactgg tcaacccaag
     3541 gttaggaagt tttttctatt ggtatttaaa gtccgagtct gaggacaagc catatttaga
     3601 ccagatttta agttcctttt ggagtagact ggataaaaaa tctcggaata tactgaacga
     3661 tcaagtcagg ctaataaatg tgctcaggga atgttgtgaa acaattaaga ggctgaaaga
     3721 tactacagcc aaaaagatgg aattactagt gcatttattg gagacgaaag tcaggcctct
     3781 tgtcaaagta cgaccaattg ctttaccact agaccctgat gtgttgatat gcgacgtttg
     3841 tcctgaaact tcgaaggtat ttaaaagctc tttatcgcca ctaaaaataa cattcaaaac
     3901 gaccttaaac caaccatatc acttaatgtt taaggttggc gatgatttga gacaagatca
     3961 actggtagta cagatcataa gtttaatgaa tgagttatta aaaaatgaaa acgtggactt
     4021 gaaattgaca ccatacaaaa ttttggcaac aggaccacaa gaaggtgcca ttgaatttat
     4081 ccctaatgac acattagcta gcatactaag caaatatcac ggtattcttg gctaccttaa
     4141 actccactat cctgatgaga acgcgacatt aggcgtacaa ggctgggttt tagataattt
     4201 tgtcaaatct tgtgctggct attgtgttat tacatacatc ttaggtgtcg gcgataggca
     4261 tttagacaac ttactagtca cgccagatgg gcactttttt cacgcagact ttggttatat
     4321 cttgggtcag gaccccaaac cttttccgcc attaatgaaa ttgccccctc aaattataga
     4381 ggcgtttggg ggtgcagaat catcaaatta tgataaattt cgcagctact gttttgtggc
     4441 atattcgatt ttaagaagga atgcaggctt aatcttaaac ctgtttgaat taatgaagac
     4501 ttcgaacata cctgatatca gaatagatcc caatggtgct atattacgtg taagagagag
     4561 atttaatttg aatatgtccg aagaagatgc cacagtgcat tttcagaatc taatcaatga
     4621 tagtgtaaat gctttgctgc ctatcgtgat tgatcattta cataatctgg cacaatactg
     4681 gcggaccgtc gaggactaca aagaccatga cggtgattat aaagatcatg atatcgatta
     4741 caaggatgac gatgacaagt agaattcgtg agctcgcgaa tttcttatga tttatgattt
     4801 ttattattaa ataagttata aaaaaaataa gtgtatacaa attttaaagt gactcttagg
     4861 ttttaaaacg aaaattctta ttcttgagta actctttcct gtaggtcagg ttgctttctc
     4921 aggtatagca tgaggtcgct cttattgacc acacctctac cggcatgccg agcaaatgcc
     4981 tgcaaatcgc tccccgcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg
     5041 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5101 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5161 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5221 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5281 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5341 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5401 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5461 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5521 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5581 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5641 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5701 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5761 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5821 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5881 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5941 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6001 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6061 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6121 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6181 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6241 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6301 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6361 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6421 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6481 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6541 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6601 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6661 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6721 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6781 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6841 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6901 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6961 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7021 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7081 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7141 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7201 atttccccga aaagtgccac ctgggtcctt ttcatcacgt gctataaaaa taattataat
     7261 ttaaattttt taatataaat atataaatta aaaatagaaa gtaaaaaaag aaattaaaga
     7321 aaaaatagtt tttgttttcc gaagatgtaa aagactctag ggggatcgcc aacaaatact
     7381 accttttatc ttgctcttcc tgctctcagg tattaatgcc gaattgtttc atcttgtctg
     7441 tgtagaagac cacacacgaa aatcctgtga ttttacattt tacttatcgt taatcgaatg
     7501 tatatctatt taatctgctt ttcttgtcta ataaatatat atgtaaagta cgctttttgt
     7561 tgaaattttt taaacctttg tttatttttt tttcttcatt ccgtaactct tctaccttct
     7621 ttatttactt tctaaaatcc aaatacaaaa cataaaaata aataaacaca gagtaaattc
     7681 ccaaattatt ccatcattaa aagatacgag gcgcgtgtaa gttacaggca agcgatccgt
     7741 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     7801 tcgtc
//
