LOCUS       sequence-472489-        7989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ScVPS38-EGFP under its own promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7989)
  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 7989)
  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: pYO764
FEATURES             Location/Qualifiers
     source          1..7989
                     /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(4267..4269,4270..4983)
                     /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(4267..4269,4270..4980)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4309..4330)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4570..4589)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4917..4938
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      4990..5177
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5045..5069)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(5216..5236)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5216..5234)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5255..5271)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5255..5271)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5268..5290)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5279..5295
                     /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(5303..5309,5310..5327,5328..5333))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5348..5369
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5486..5503)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5657..6245)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5737..5756)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6416..7207,7208..7276))
                     /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     7039..7058
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7277..7381)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7418..7921
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7962..7980
                     /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 atttactttg aaaaaccttc tagcgtaaat gatgttatta ataggctcaa gctaaaacct
     3961 ttgaatatag agatattact tagacaagcc ggcgaatcca caaagcacag agaatatgta
     4021 aacagccagt tgggttatta ccttttattt ctccatttga cagccatcca aattttcaag
     4081 gctcctttac cctatagact aatgtattat ggtagtacat cagtgatcga cagtcaatat
     4141 ccgttgtatt tcactgatca aatgatttct aagcatcaag caaagctgat taaagccata
     4201 cattatttta atgctgatat attgcaattt aaacagattc tggagaatta ccgtccaaca
     4261 gtcgaggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     4321 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     4381 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     4441 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     4501 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     4561 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     4621 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     4681 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     4741 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     4801 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     4861 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     4921 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     4981 taagagctcg cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa
     5041 ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg
     5101 agtaactctt tcctgtaggt caggttgctt tctcaggtat agcatgaggt cgctcttatt
     5161 gaccacacct ctaccggcag cggccgccac cgcggtggag ctccagcttt tgttcccttt
     5221 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5281 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5341 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5401 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5461 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5521 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5581 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5641 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5701 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5761 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5821 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5881 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5941 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6001 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6061 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6121 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6181 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6241 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6301 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6361 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6421 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6481 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6541 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6601 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6661 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6721 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6781 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6841 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6901 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6961 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7021 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7081 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7141 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7201 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7261 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7321 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7381 gcacatttcc ccgaaaagtg ccacctgggt ccttttcatc acgtgctata aaaataatta
     7441 taatttaaat tttttaatat aaatatataa attaaaaata gaaagtaaaa aaagaaatta
     7501 aagaaaaaat agtttttgtt ttccgaagat gtaaaagact ctagggggat cgccaacaaa
     7561 tactaccttt tatcttgctc ttcctgctct caggtattaa tgccgaattg tttcatcttg
     7621 tctgtgtaga agaccacaca cgaaaatcct gtgattttac attttactta tcgttaatcg
     7681 aatgtatatc tatttaatct gcttttcttg tctaataaat atatatgtaa agtacgcttt
     7741 ttgttgaaat tttttaaacc tttgtttatt tttttttctt cattccgtaa ctcttctacc
     7801 ttctttattt actttctaaa atccaaatac aaaacataaa aataaataaa cacagagtaa
     7861 attcccaaat tattccatca ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat
     7921 ccgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     7981 cctttcgtc
//
