LOCUS       sequence-472483-        7640 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ScVPS38 delta BARA2 (320-440)-EGFP under its own 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7640)
  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 7640)
  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: pYO766
FEATURES             Location/Qualifiers
     source          1..7640
                     /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(3907..3909,3910..3912,3913..4626)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3907..3909,3910..3912,3913..4623)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3952..3973)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4213..4232)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4560..4581
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      4633..4820
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4688..4712)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(4867..4887)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4867..4885)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4906..4922)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4906..4922)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4919..4941)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4930..4946
                     /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(4954..4960,4961..4978,4979..4984))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4999..5020
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5137..5154)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5308..5896)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5388..5407)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6067..6858,6859..6927))
                     /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     6690..6709
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6928..7032)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7069..7572
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7613..7631
                     /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 atgtttcatg
     2941 ggcatgaaac ggttcttact cagtcgaaga cagagacacc ttcgtatgat ttgctttcac
     3001 aatataagtc tctttagagc taatggtgac tccaaattga tcaaggaata tggtgatgga
     3061 tttataccat gtttctttat cttggaatcc ataagagggg agctgcttta tgtgagcgag
     3121 gtacaatcag gttctttgcg taaactctct tttcaggaat taccaaaact aactggtgca
     3181 tctactatga ttgtcctaaa attggtagga ctagttccga gtgatatttt gtgtaccata
     3241 agttcagata aaaatggtat cattgacgat aaatggtgcg ttttatgtac ctacacgata
     3301 gatcttaaca agttacaacc cataaatgaa gacacagttc taattacagg caccaacgca
     3361 ccagtacttg atttaataga tggatcctac acattggctg cagaaaaaat taagccacta
     3421 aagggccttg ttagttctca caaaaggaat ataagtcaag tcaagatcaa attttcattg
     3481 gcatatagct ctttactaaa attaaacaaa ctattggagt attcttccca agtccatgag
     3541 gaaatcaatg aaatttccag caaaatagaa gatgactttt tgtcactcaa gaatcaaaat
     3601 cattggtata tgagaacagt acagaagtcc atagaaacgc tagaaaagga agtcctacaa
     3661 aggaaaaaaa gcaagaaaaa tatagaaatg gcgcaactcg agagtaatga tactatcaac
     3721 cattctaaaa cagaactgtc tctgatgtcg caggacgagt ctataaatga tgactatgga
     3781 agcatatatt ctagatttgt gcaaataaaa gataggttgg accaactgag gtttaagaaa
     3841 ctgtatcagc tgatagggat cttccattca acggatttat ttaattcaga tagaggctat
     3901 gtcgagatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     3961 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     4021 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     4081 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     4141 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     4201 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     4261 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     4321 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     4381 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     4441 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     4501 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     4561 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     4621 aagtaagagc tcgcgaattt cttatgattt atgattttta ttattaaata agttataaaa
     4681 aaaataagtg tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc
     4741 ttgagtaact ctttcctgta ggtcaggttg ctttctcagg tatagcatga ggtcgctctt
     4801 attgaccaca cctctaccgg caggccggcc gcggccgcca ccgcggtgga gctccagctt
     4861 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     4921 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4981 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5041 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5101 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5161 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5221 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5281 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5341 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5401 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5461 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5521 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5581 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5641 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5701 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5761 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5821 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5881 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5941 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6001 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6061 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6121 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6181 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6241 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6301 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6361 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6421 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6481 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6541 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6601 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6661 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6721 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6781 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6841 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6901 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6961 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7021 aggggttccg cgcacatttc cccgaaaagt gccacctggg tccttttcat cacgtgctat
     7081 aaaaataatt ataatttaaa ttttttaata taaatatata aattaaaaat agaaagtaaa
     7141 aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac tctaggggga
     7201 tcgccaacaa atactacctt ttatcttgct cttcctgctc tcaggtatta atgccgaatt
     7261 gtttcatctt gtctgtgtag aagaccacac acgaaaatcc tgtgatttta cattttactt
     7321 atcgttaatc gaatgtatat ctatttaatc tgcttttctt gtctaataaa tatatatgta
     7381 aagtacgctt tttgttgaaa ttttttaaac ctttgtttat ttttttttct tcattccgta
     7441 actcttctac cttctttatt tactttctaa aatccaaata caaaacataa aaataaataa
     7501 acacagagta aattcccaaa ttattccatc attaaaagat acgaggcgcg tgtaagttac
     7561 aggcaagcga tccgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     7621 tatcacgagg ccctttcgtc
//
