LOCUS       sequence-472487-        7536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ScVPS38 delta C2 (1-154) -EGFP under its own promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7536)
  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 7536)
  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: pYO765
FEATURES             Location/Qualifiers
     source          1..7536
                     /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             join(3806..3808,3809..4522)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(3806..3808,3809..4519)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3848..3869)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4109..4128)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4456..4477
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      4529..4716
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4584..4608)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(4763..4783)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4763..4781)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4802..4818)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4802..4818)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4815..4837)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4826..4842
                     /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(4850..4856,4857..4874,4875..4880))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4895..4916
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5033..5050)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5204..5792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5284..5303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5963..6754,6755..6823))
                     /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     6586..6605
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6824..6928)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6965..7468
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7509..7527
                     /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 tgggtaccgc gttgtgagag agtgtgctcc ttggctagac acttaggttc
     2041 cttgtttagt gacgccgttc aaactgcatc cgttcaatgg ttcgaaagaa ctctggatga
     2101 ttctgctatt agaagaattt ctttaccaag tgcattttta accgcagata ttctattatc
     2161 tactttgttg aacatctcat ccggtttagt tgtgtatcca aaggttatcg aaaggagaat
     2221 taagggtgaa ctacctttta tggctactga aaatatcatc atggctatgg tagaaaagaa
     2281 tgcctccaga caagaagtac atgagcgtat tagagtgctc tctcatcaag ccgcagcagt
     2341 agtcaaggaa gaaggtgggg aaaatgattt aattgaacga gtaaagaggg atgaattttt
     2401 caagcctatc tgggaagaat tagattcttt actggaacca tccacttttg ttggtagagc
     2461 tccacaacaa gttgagaaat ttgttcaaaa agacgttaac aatgctttac aacctttcca
     2521 aaagtaccta aacgatgaac aagtcaagtt aaatgtttag aagcctttac atgtacaaat
     2581 aatatgttta ttaccctatt atatgttact aatgaaaaaa ataacaattc tttatttatg
     2641 tacaatgcaa gtagaattcc ctaaatagaa gttaactatt tttattttac tgggatatat
     2701 ttttcatctt cttccttcat actcctttca tttttcttgt ttttttggct tacggatttt
     2761 ggcaatgttt atactttaga aaaaaaggaa actcataatg ctggcatact tgaaaaacca
     2821 tacggaaaga tcgtagttta tttttgtaat aagacactga agtcagctgt ttgaaagagt
     2881 atattgaatt gatggtttta cctattaggg atagtaatca taatttaaaa atgtttgatc
     2941 catgaaaatt aagccactaa agggccttgt tagttctcac aaaaggaata taagtcaagt
     3001 caagatcaaa ttttcattgg catatagctc tttactaaaa ttaaacaaac tattggagta
     3061 ttcttcccaa gtccatgagg aaatcaatga aatttccagc aaaatagaag atgacttttt
     3121 gtcactcaag aatcaaaatc attggtatat gagaacagta cagaagtcca tagaaacgct
     3181 agaaaaggaa gtcctacaaa ggaaaaaaag caagaaaaat atagaaatgg cgcaactcga
     3241 gagtaatgat actatcaacc attctaaaac agaactgtct ctgatgtcgc aggacgagtc
     3301 tataaatgat gactatggaa gcatatattc tagatttgtg caaataaaag ataggttgga
     3361 ccaactgagg tttaagaaac tgtatcagct gatagggatc ttccattcaa cggatttatt
     3421 taattcagat agaggctata tttactttga aaaaccttct agcgtaaatg atgttattaa
     3481 taggctcaag ctaaaacctt tgaatataga gatattactt agacaagccg gcgaatccac
     3541 aaagcacaga gaatatgtaa acagccagtt gggttattac cttttatttc tccatttgac
     3601 agccatccaa attttcaagg ctcctttacc ctatagacta atgtattatg gtagtacatc
     3661 agtgatcgac agtcaatatc cgttgtattt cactgatcaa atgatttcta agcatcaagc
     3721 aaagctgatt aaagccatac attattttaa tgctgatata ttgcaattta aacagattct
     3781 ggagaattac cgtccaacag tcgaggtgag caagggcgag gagctgttca ccggggtggt
     3841 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     3901 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3961 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     4021 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     4081 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     4141 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     4201 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     4261 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     4321 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     4381 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     4441 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     4501 catggacgag ctgtacaagt aagagctcgc gaatttctta tgatttatga tttttattat
     4561 taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa
     4621 acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata
     4681 gcatgaggtc gctcttattg accacacctc taccggcagg ccggccgcgg ccgccaccgc
     4741 ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     4801 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4861 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4921 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4981 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5041 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5101 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5161 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5221 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5281 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5341 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5401 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5461 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5521 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5581 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5641 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5701 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5761 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5821 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5881 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5941 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6001 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6061 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6121 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6181 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6241 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6301 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6361 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6421 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6481 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6541 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6601 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6661 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6721 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6781 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6841 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6901 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcct
     6961 tttcatcacg tgctataaaa ataattataa tttaaatttt ttaatataaa tatataaatt
     7021 aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc cgaagatgta
     7081 aaagactcta gggggatcgc caacaaatac taccttttat cttgctcttc ctgctctcag
     7141 gtattaatgc cgaattgttt catcttgtct gtgtagaaga ccacacacga aaatcctgtg
     7201 attttacatt ttacttatcg ttaatcgaat gtatatctat ttaatctgct tttcttgtct
     7261 aataaatata tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt gtttattttt
     7321 ttttcttcat tccgtaactc ttctaccttc tttatttact ttctaaaatc caaatacaaa
     7381 acataaaaat aaataaacac agagtaaatt cccaaattat tccatcatta aaagatacga
     7441 ggcgcgtgta agttacaggc aagcgatccg tctaagaaac cattattatc atgacattaa
     7501 cctataaaaa taggcgtatc acgaggccct ttcgtc
//
