LOCUS       sequence-472643-        9090 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of FL human VPS15 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9090)
  AUTHORS   Tremel S, Ohashi Y, Morado DR, Bertram J, Perisic O, Brandt LTL, von
            Wrisberg MK, Chen ZA, Maslen SL, Kovtun O, Skehel M, Rappsilber J, 
            Lang K, Munro S, Briggs JAG, Williams RL
  TITLE     Structural basis for VPS34 kinase activation by Rab1 and Rab5 on 
            membranes.
  JOURNAL   Nat Commun. 2021 Mar 10;12(1):1564. doi: 10.1038/s41467-021-21695-2.
  PUBMED    33692360
REFERENCE   2  (bases 1 to 9090)
  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: "Nat 
            Commun."; date: "2021-03-10"; volume: "12(1):1564. doi"; pages: " 
            10.1038/s41467-021-21695-2"
COMMENT     Sequence Label: pYO350
FEATURES             Location/Qualifiers
     source          1..9090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(5098..5115)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5104..5328
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5374..5802
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5461..5480)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5671..5692
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5811..5831)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5816..6145
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5996..6131
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6058..6077
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6193..6240
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6259..6633
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    6763..6884
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6800..6819)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6854..6873
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6933..6949)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6933..6949)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6946..6968)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6957..6973
                     /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(6981..6987,6988..7005,7006..7011))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7026..7047
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7164..7181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7335..7923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7415..7434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8094..8885,8886..8954))
                     /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     8717..8736
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8955..9059)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttatgggaa atcagcttgc tggcattgct ccctcccaga
     1021 tcctttctgt agagagttat ttttcagata ttcatgactt tgaatatgat aaaagcctgg
     1081 ggagtactcg gttttttaaa gttgctcgag ccaagcaccg agaaggcctg gtcgttgtga
     1141 aggtttttgc aattcaggat cccacattgc ctttaaccag ctataaacaa gagctggagg
     1201 aactgaaaat caggcttaat tctgcacaga attgtctacc tttccagaaa gcatcagaaa
     1261 aagcatctga gaaagcagct atgctcttta ggcagtatgt gcgagacaat ctctatgatc
     1321 gcatcagtac ccgtccattc ttgaataaca ttgagaagcg ctggattgct ttccagatcc
     1381 tgacagctgt ggaccaagca cacaaatctg gagttcgtca tggggacatc aagactgaga
     1441 atgtgatggt caccagttgg aattgggttc ttctaactga ttttgccagt tttaagccca
     1501 cttatcttcc agaagacaac ccggcagatt tcaattattt ctttgacaca tcacggagga
     1561 gaacttgcta tattgctcct gaacgttttg ttgatggtgg gatgtttgcc actgagttag
     1621 aatatatgag agatccttca actccgcttg tagacttaaa tagcaatcag agaacaagag
     1681 gagagttgaa gagagcaatg gacatctttt cagcaggttg tgtgatagct gagcttttta
     1741 cagaaggtgt accattattt gatctctctc aacttttggc ttatagaaat ggacattttt
     1801 tccctgaaca agtgctaaat aaaattgaag atcacagtat cagagaattg gtaactcaga
     1861 tgattcaccg tgagccagat aaacgtttag aggcagaaga ttacttaaaa cagcagcgtg
     1921 gcaatgcctt tcctgaaata ttttacactt ttcttcagcc ctacatggcc cagtttgcca
     1981 aggaaacgtt tctttctgca gatgagcgta ttctggttat acggaaggat ttgggcaaca
     2041 ttattcacaa tctctgtgga catgatctgc cagaaaaagc cgaaggagag cctaaggaaa
     2101 atgggctggt tatcttggta tctgttataa catcctgcct acagaccctt aaatactgtg
     2161 attccaaact agctgctttg gaactgattc ttcatttggc tccaagatta agtgttgaaa
     2221 tccttttgga tcgtattact ccatatcttt tgcatttcag caatgactct gttcctaggg
     2281 tgagggctga agccttgagg acgttgacca aagttcttgc tctcgtcaaa gaggttcctc
     2341 gtaatgatat caatatttat ccggaataca ttctgccagg catagcccac ttagcccaag
     2401 atgatgctac tatcgttaga ctagcctatg ctgaaaacat agctctgctg gcagaaacag
     2461 ctctgagatt cctggaatta gtacagttaa aaaatcttaa tatggaaaat gaccccaata
     2521 atgaagaaat agatgaggtt acacatccaa atggaaatta tgacacagag ctgcaagcct
     2581 tacatgaaat ggtccagcag aaagttgtta ctttgctaag tgaccctgaa aatattgtaa
     2641 aacaaacctt gatggaaaat ggaataacac ggctgtgtgt attctttgga cgtcagaaag
     2701 ccaacgatgt tttgttgtcc cacatgatta ctttcctaaa tgataagaat gattggcatc
     2761 tacgtggagc attttttgat agtatagttg gtgttgctgc ctatgttggc tggcaaagct
     2821 cctcaattct caagcctctg ctgcaacaag gtcttagtga tgctgaggaa tttgtcattg
     2881 tgaaagctct ttatgccctt acttgtatgt gccagttagg actgctacaa aaaccccatg
     2941 tttacgaatt tgccagtgat attgccccct tcctgtgtca tcccaattta tggatacgtt
     3001 atggtgccgt gggatttatc acagtggtag ctcgtcaaat aagtacagct gatgtctact
     3061 gtaaactgat gccttatctt gacccatata ttacccaacc aataatacag attgaaagaa
     3121 aacttgttct gctcagtgtt ttaaaggaac cagtaagtcg ttctatattt gattatgctt
     3181 tgaggtctaa agatattact agcttgttca gacatcttca catgcgtcag aagaaacgaa
     3241 atggttctct tcccgactgc cctccgccag aggatcctgc catagcacag cttctgaaga
     3301 agttgctctc acagggaatg acagaggaag aggaagacaa acttctggca ctgaaagact
     3361 tcatgatgaa atctaataaa gcaaaggcca atatagtgga ccagagccat cttcatgata
     3421 gtagtcagaa aggtgtaatt gacttggcag ctttaggcat aactgggaga caagttgatc
     3481 ttgttaaaac caaacaagaa ccagatgaca aacgggccag aaaacatgta aaacaagact
     3541 caaatgtaaa tgaagaatgg aaaagcatgt ttgggtcact ggacccacca aacatgccac
     3601 aggccctacc taaagggagt gatcaggagg tgattcagac tgggaaacct cctcgttccg
     3661 agtcctctgc tggcatttgt gtccctttgt caacttcttc acaggttcca gaagtgacaa
     3721 ctgtccaaaa taaaaaacca gtaataccgg ttttaagtag tacaatctta ccatccacct
     3781 atcagattcg aattacaact tgtaaaactg aacttcagca actcatccag caaaagcggg
     3841 agcagtgcaa tgctgagaga atagctaagc agatgatgga aaatgctgaa tgggagagta
     3901 aaccaccacc acctggatgg cgtcctaaag ggctgttagt tgcccatctt catgagcata
     3961 aatctgctgt gaatcgaatt agagtctctg atgaacactc actttttgca acatgttcaa
     4021 atgatggcac agtgaaaatc tggaacagtc aaaagatgga ggggaagacc accactacca
     4081 gatctattct tacatacagc cgaattggag gacgagtcaa gacgctcaca ttctgccaag
     4141 gctcccacta tttagccata gcatctgata atggtgctgt ccagcttctt ggaattgagg
     4201 cttctaagct gcccaagtct cctaaaatcc atcctctaca aagcagaatt ctagatcaga
     4261 aggaggacgg ttgtgttgtg gatatgcatc acttcaactc tggagcacag tctgttcttg
     4321 cctatgccac tgtgaatggc tctctggttg gctgggacct taggtcttca agcaatgcgt
     4381 ggactttaaa gcatgattta aagtcgggcc tcatcacttc ctttgctgtg gacatccacc
     4441 aatgctggct ctgcattggt acaagcagtg gcaccatggc ttgttgggac atgaggttcc
     4501 agttgccaat ttcaagtcac tgtcatcctt ccagggctcg aatcagacgc ctctcaatgc
     4561 accctctgta tcagtcctgg gtgattgcag ctgttcaggg caacaacgaa gtgtccatgt
     4621 gggacatgga gactggtgac agaagattta ctctctgggc cagcagtgca ccaccacttt
     4681 ctgaattaca gccttctcct catagcgtcc atggtatcta ctgtagtcct gcagatggaa
     4741 atcctatcct actaacagct ggctcagata tgaaaataag gttttgggac ttggcttacc
     4801 cagaaaggtc ctatgttgtt gcaggaagta ctagttcccc atctgtgtcc tactacagga
     4861 aaataattga aggcactgaa gttgtccagg aaattcagaa taagcagaaa gtaggaccaa
     4921 gtgatgacac ccctcgaagg ggcccagagt ccctgcccgt gggacatcat gacatcatca
     4981 ctgatgtcgc cacattccag accacacagg gcttcatcgt aactgcttct agagatggga
     5041 ttgtgaaggt gtggaaataa gtcgagtcta gagggcccgt ttaaacccgc tgatcagcct
     5101 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     5161 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     5221 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     5281 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     5341 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     5401 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     5461 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     5521 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     5581 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     5641 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     5701 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     5761 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     5821 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5881 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5941 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     6001 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6061 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     6121 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     6181 tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
     6241 aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc
     6301 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg
     6361 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag
     6421 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
     6481 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg
     6541 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac
     6601 tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc
     6661 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     6721 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     6781 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6841 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     6901 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6961 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     7021 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     7081 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     7141 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7201 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7261 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7321 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7381 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7441 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7501 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     7561 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7621 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7681 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7741 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     7801 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7861 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7921 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7981 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     8041 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     8101 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     8161 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     8221 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     8281 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8341 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8401 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8461 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8521 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8581 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8641 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8701 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8761 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8821 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8881 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8941 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     9001 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     9061 acatttcccc gaaaagtgcc acctgacgtc
//
