LOCUS       sequence-472491-        6887 bp    DNA     circular SYN 13-MAY-2025
DEFINITION  Expression of ScVPS38 delta C2 (1-154)-x3FLAG under its own 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6887)
  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 6887)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 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: pYO760
FEATURES             Location/Qualifiers
     source          1..6887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..52)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     152..171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        195..410
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             411..1214
                     /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(1345..1800)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1482..1503)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1694..1713
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1926..1948
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1940..1957
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1941..1957
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1967..1986
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1967..1985
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3807..3872
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3889..4076
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3944..3968)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(4115..4135)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4115..4133)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4154..4170)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4154..4170)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4167..4189)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4178..4194
                     /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(4202..4208,4209..4226,4227..4232))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4247..4268
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4385..4402)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4556..5144)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4636..4655)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5315..6106,6107..6175))
                     /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     5938..5957
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6176..6280)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6317..6820
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6861..6879
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
       61 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
      121 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
      181 caccatacca ccttttcaat tcatcatttt ttttttattc ttttttttga tttcggtttc
      241 cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
      301 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
      361 tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag
      421 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
      481 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
      541 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
      601 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
      661 acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc
      721 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
      781 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
      841 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
      901 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
      961 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     1021 tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     1081 tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     1141 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     1201 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     1261 gagcttcaat ttaattatat cagttattac cctatgcggt gtgaaatacc gcacagatgc
     1321 gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt
     1381 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     1441 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     1501 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     1561 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     1621 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     1681 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     1741 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     1801 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     1861 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     1921 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact
     1981 atagggcgaa ttgggtaccg cgttgtgaga gagtgtgctc cttggctaga cacttaggtt
     2041 ccttgtttag tgacgccgtt caaactgcat ccgttcaatg gttcgaaaga actctggatg
     2101 attctgctat tagaagaatt tctttaccaa gtgcattttt aaccgcagat attctattat
     2161 ctactttgtt gaacatctca tccggtttag ttgtgtatcc aaaggttatc gaaaggagaa
     2221 ttaagggtga actacctttt atggctactg aaaatatcat catggctatg gtagaaaaga
     2281 atgcctccag acaagaagta catgagcgta ttagagtgct ctctcatcaa gccgcagcag
     2341 tagtcaagga agaaggtggg gaaaatgatt taattgaacg agtaaagagg gatgaatttt
     2401 tcaagcctat ctgggaagaa ttagattctt tactggaacc atccactttt gttggtagag
     2461 ctccacaaca agttgagaaa tttgttcaaa aagacgttaa caatgcttta caacctttcc
     2521 aaaagtacct aaacgatgaa caagtcaagt taaatgttta gaagccttta catgtacaaa
     2581 taatatgttt attaccctat tatatgttac taatgaaaaa aataacaatt ctttatttat
     2641 gtacaatgca agtagaattc cctaaataga agttaactat ttttatttta ctgggatata
     2701 tttttcatct tcttccttca tactcctttc atttttcttg tttttttggc ttacggattt
     2761 tggcaatgtt tatactttag aaaaaaagga aactcataat gctggcatac ttgaaaaacc
     2821 atacggaaag atcgtagttt atttttgtaa taagacactg aagtcagctg tttgaaagag
     2881 tatattgaat tgatggtttt acctattagg gatagtaatc ataatttaaa aatgtttgat
     2941 ccatgaaaat taagccacta aagggccttg ttagttctca caaaaggaat ataagtcaag
     3001 tcaagatcaa attttcattg gcatatagct ctttactaaa attaaacaaa ctattggagt
     3061 attcttccca agtccatgag gaaatcaatg aaatttccag caaaatagaa gatgactttt
     3121 tgtcactcaa gaatcaaaat cattggtata tgagaacagt acagaagtcc atagaaacgc
     3181 tagaaaagga agtcctacaa aggaaaaaaa gcaagaaaaa tatagaaatg gcgcaactcg
     3241 agagtaatga tactatcaac cattctaaaa cagaactgtc tctgatgtcg caggacgagt
     3301 ctataaatga tgactatgga agcatatatt ctagatttgt gcaaataaaa gataggttgg
     3361 accaactgag gtttaagaaa ctgtatcagc tgatagggat cttccattca acggatttat
     3421 ttaattcaga tagaggctat atttactttg aaaaaccttc tagcgtaaat gatgttatta
     3481 ataggctcaa gctaaaacct ttgaatatag agatattact tagacaagcc ggcgaatcca
     3541 caaagcacag agaatatgta aacagccagt tgggttatta ccttttattt ctccatttga
     3601 cagccatcca aattttcaag gctcctttac cctatagact aatgtattat ggtagtacat
     3661 cagtgatcga cagtcaatat ccgttgtatt tcactgatca aatgatttct aagcatcaag
     3721 caaagctgat taaagccata cattatttta atgctgatat attgcaattt aaacagattc
     3781 tggagaatta ccgtccaaca gtcgaggact acaaagacca tgacggtgat tataaagatc
     3841 atgatatcga ttacaaggat gacgatgaca agtagaattc gtgagctcgc gaatttctta
     3901 tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa
     3961 agtgactctt aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc
     4021 aggttgcttt ctcaggtata gcatgaggtc gctcttattg accacacctc taccggcagc
     4081 ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct
     4141 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4201 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4261 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4321 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4381 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4441 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4501 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4561 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4621 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4681 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4741 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4801 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4861 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4921 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4981 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5041 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5101 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5161 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5221 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5281 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5341 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5401 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5461 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5521 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5581 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5641 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5701 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5761 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5821 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5881 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5941 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6001 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6061 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6121 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6181 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6241 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6301 cacctgggtc cttttcatca cgtgctataa aaataattat aatttaaatt ttttaatata
     6361 aatatataaa ttaaaaatag aaagtaaaaa aagaaattaa agaaaaaata gtttttgttt
     6421 tccgaagatg taaaagactc tagggggatc gccaacaaat actacctttt atcttgctct
     6481 tcctgctctc aggtattaat gccgaattgt ttcatcttgt ctgtgtagaa gaccacacac
     6541 gaaaatcctg tgattttaca ttttacttat cgttaatcga atgtatatct atttaatctg
     6601 cttttcttgt ctaataaata tatatgtaaa gtacgctttt tgttgaaatt ttttaaacct
     6661 ttgtttattt ttttttcttc attccgtaac tcttctacct tctttattta ctttctaaaa
     6721 tccaaataca aaacataaaa ataaataaac acagagtaaa ttcccaaatt attccatcat
     6781 taaaagatac gaggcgcgtg taagttacag gcaagcgatc cgtctaagaa accattatta
     6841 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgt
//
