LOCUS       sequence-472853-        9477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of FL human VPS15-ZZ in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9477)
  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 9477)
  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: pYO1015
FEATURES             Location/Qualifiers
     source          1..9477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          664..1680
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1604..1626
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1688..1707
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1754..1763
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5855..5875
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             5897..5917
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             5999..6019
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             6035..6208
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             6209..6382
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     primer_bind     complement(6499..6518)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6564..6619
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6618..6637)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6980..6996)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6980..6996)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6993..7015)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7004..7020
                     /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(7028..7034,7035..7052,7053..7058))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7073..7094
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7153..7348
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7199..7334
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    7354..7488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7391..7410)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7445..7464
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7556..7573)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7727..8315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7807..7826)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8486..9277,9278..9346))
                     /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     9109..9128
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9347..9451)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gcgccrgggg gggggggggg gggsgggggg
      541 gggggsgggg sgrggsggrg rggkgcggcg gcagccaatc agagcggcgc gctccgaaag
      601 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
      661 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
      721 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      781 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
      841 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
      961 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     1021 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     1081 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
     1141 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
     1201 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggkgggggt gccgggcggg
     1261 gcggggccgc ckcgggccgg ggggggctcg ggggrggggc gcggcggccc ccggagcgcc
     1321 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     1381 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     1441 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1501 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     1561 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     1621 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     1681 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attgctcgat
     1741 cgacggtatc gatgccacca tgggaaatca gcttgctggc attgctccct cccagatcct
     1801 ttctgtagag agttattttt cagatattca tgactttgaa tatgataaaa gcctggggag
     1861 tactcggttt tttaaagttg ctcgagccaa gcaccgagaa ggcctggtcg ttgtgaaggt
     1921 ttttgcaatt caggatccca cattgccttt aaccagctat aaacaagagc tggaggaact
     1981 gaaaatcagg cttaattctg cacagaattg tctacctttc cagaaagcat cagaaaaagc
     2041 atctgagaaa gcagctatgc tctttaggca gtatgtgcga gacaatctct atgatcgcat
     2101 cagtacccgt ccattcttga ataacattga gaagcgctgg attgctttcc agatcctgac
     2161 agctgtggac caagcacaca aatctggagt tcgtcatggg gacatcaaga ctgagaatgt
     2221 gatggtcacc agttggaatt gggttcttct aactgatttt gccagtttta agcccactta
     2281 tcttccagaa gacaacccgg cagatttcaa ttatttcttt gacacatcac ggaggagaac
     2341 ttgctatatt gctcctgaac gttttgttga tggtgggatg tttgccactg agttagaata
     2401 tatgagagat ccttcaactc cgcttgtaga cttaaatagc aatcagagaa caagaggaga
     2461 gttgaagaga gcaatggaca tcttttcagc aggttgtgtg atagctgagc tttttacaga
     2521 aggtgtacca ttatttgatc tctctcaact tttggcttat agaaatggac attttttccc
     2581 tgaacaagtg ctaaataaaa ttgaagatca cagtatcaga gaattggtaa ctcagatgat
     2641 tcaccgtgag ccagataaac gtttagaggc agaagattac ttaaaacagc agcgtggcaa
     2701 tgcctttcct gaaatatttt acacttttct tcagccctac atggcccagt ttgccaagga
     2761 aacgtttctt tctgcagatg agcgtattct ggttatacgg aaggatttgg gcaacattat
     2821 tcacaatctc tgtggacatg atctgccaga aaaagccgaa ggagagccta aggaaaatgg
     2881 gctggttatc ttggtatctg ttataacatc ctgcctacag acccttaaat actgtgattc
     2941 caaactagct gctttggaac tgattcttca tttggctcca agattaagtg ttgaaatcct
     3001 tttggatcgt attactccat atcttttgca tttcagcaat gactctgttc ctagggtgag
     3061 ggctgaagcc ttgaggacgt tgaccaaagt tcttgctctc gtcaaagagg ttcctcgtaa
     3121 tgatatcaat atttatccgg aatacattct gccaggcata gcccacttag cccaagatga
     3181 tgctactatc gttagactag cctatgctga aaacatagct ctgctggcag aaacagctct
     3241 gagattcctg gaattagtac agttaaaaaa tcttaatatg gaaaatgacc ccaataatga
     3301 agaaatagat gaggttacac atccaaatgg aaattatgac acagagctgc aagccttaca
     3361 tgaaatggtc cagcagaaag ttgttacttt gctaagtgac cctgaaaata ttgtaaaaca
     3421 aaccttgatg gaaaatggaa taacacggct gtgtgtattc tttggacgtc agaaagccaa
     3481 cgatgttttg ttgtcccaca tgattacttt cctaaatgat aagaatgatt ggcatctacg
     3541 tggagcattt tttgatagta tagttggtgt tgctgcctat gttggctggc aaagctcctc
     3601 aattctcaag cctctgctgc aacaaggtct tagtgatgct gaggaatttg tcattgtgaa
     3661 agctctttat gcccttactt gtatgtgcca gttaggactg ctacaaaaac cccatgttta
     3721 cgaatttgcc agtgatattg cccccttcct gtgtcatccc aatttatgga tacgttatgg
     3781 tgccgtggga tttatcacag tggtagctcg tcaaataagt acagctgatg tctactgtaa
     3841 actgatgcct tatcttgacc catatattac ccaaccaata atacagattg aaagaaaact
     3901 tgttctgctc agtgttttaa aggaaccagt aagtcgttct atatttgatt atgctttgag
     3961 gtctaaagat attactagct tgttcagaca tcttcacatg cgtcagaaga aacgaaatgg
     4021 ttctcttccc gactgccctc cgccagagga tcctgccata gcacagcttc tgaagaagtt
     4081 gctctcacag ggaatgacag aggaagagga agacaaactt ctggcactga aagacttcat
     4141 gatgaaatct aataaagcaa aggccaatat agtggaccag agccatcttc atgatagtag
     4201 tcagaaaggt gtaattgact tggcagcttt aggcataact gggagacaag ttgatcttgt
     4261 taaaaccaaa caagaaccag atgacaaacg ggccagaaaa catgtaaaac aagactcaaa
     4321 tgtaaatgaa gaatggaaaa gcatgtttgg gtcactggac ccaccaaaca tgccacaggc
     4381 cctacctaaa gggagtgatc aggaggtgat tcagactggg aaacctcctc gttccgagtc
     4441 ctctgctggc atttgtgtcc ctttgtcaac ttcttcacag gttccagaag tgacaactgt
     4501 ccaaaataaa aaaccagtaa taccggtttt aagtagtaca atcttaccat ccacctatca
     4561 gattcgaatt acaacttgta aaactgaact tcagcaactc atccagcaaa agcgggagca
     4621 gtgcaatgct gagagaatag ctaagcagat gatggaaaat gctgaatggg agagtaaacc
     4681 accaccacct ggatggcgtc ctaaagggct gttagttgcc catcttcatg agcataaatc
     4741 tgctgtgaat cgaattagag tctctgatga acactcactt tttgcaacat gttcaaatga
     4801 tggcacagtg aaaatctgga acagtcaaaa gatggagggg aagaccacca ctaccagatc
     4861 tattcttaca tacagccgaa ttggaggacg agtcaagacg ctcacattct gccaaggctc
     4921 ccactattta gccatagcat ctgataatgg tgctgtccag cttcttggaa ttgaggcttc
     4981 taagctgccc aagtctccta aaatccatcc tctacaaagc agaattctag atcagaagga
     5041 ggacggttgt gttgtggata tgcatcactt caactctgga gcacagtctg ttcttgccta
     5101 tgccactgtg aatggctctc tggttggctg ggaccttagg tcttcaagca atgcgtggac
     5161 tttaaagcat gatttaaagt cgggcctcat cacttccttt gctgtggaca tccaccaatg
     5221 ctggctctgc attggtacaa gcagtggcac catggcttgt tgggacatga ggttccagtt
     5281 gccaatttca agtcactgtc atccttccag ggctcgaatc agacgcctct caatgcaccc
     5341 tctgtatcag tcctgggtga ttgcagctgt tcagggcaac aacgaagtgt ccatgtggga
     5401 catggagact ggtgacagaa gatttactct ctgggccagc agtgcaccac cactttctga
     5461 attacagcct tctcctcata gcgtccatgg tatctactgt agtcctgcag atggaaatcc
     5521 tatcctacta acagctggct cagatatgaa aataaggttt tgggacttgg cttacccaga
     5581 aaggtcctat gttgttgcag gaagtactag ttccccatct gtgtcctact acaggaaaat
     5641 aattgaaggc actgaagttg tccaggaaat tcagaataag cagaaagtag gaccaagtga
     5701 tgacacccct cgaaggggcc cagagtccct gcccgtggga catcatgaca tcatcactga
     5761 tgtcgccaca ttccagacca cacagggctt catcgtaact gcttctagag atgggattgt
     5821 gaaggtgtgg aaatctagac caactactgc ctctgaaaat ttgtactttc aaggcgagtt
     5881 gaagaccact gcttctgaaa acttgtactt ccaaggtgag ttactcgagg gtgaattgaa
     5941 gactgctgct ttggctcaac acgacgaagc tagatccgga gcaggggcgg gtgctagcga
     6001 gaacctgtac ttccagggcg cccacgatga agccgtagac aacaaattca acaaagaaca
     6061 acaaaacgcg ttctatgaga tcttacattt acctaactta aacgaagaac aacgaaacgc
     6121 cttcatccaa agtttaaaag atgacccaag ccaaagcgct aaccttttag cagaagctaa
     6181 aaagctaaat gatgctcagg cgccgaaagt agacaacaaa ttcaacaaag aacaacaaaa
     6241 cgcgttctat gagatcttac atttacctaa cttaaacgaa gaacaacgaa acgccttcat
     6301 ccaaagttta aaagatgacc caagccaaag cgctaacctt ttagcagaag ctaaaaagct
     6361 aaatgatgct caggcgccga aagtagacgc gaattcgtga gctcgcgaac tcgaggaatt
     6421 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     6481 tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     6541 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     6601 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     6661 gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa
     6721 aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt
     6781 ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt
     6841 ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc
     6901 ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc
     6961 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     7021 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     7081 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     7141 gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     7201 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     7261 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     7321 cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt
     7381 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     7441 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat
     7501 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     7561 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     7621 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     7681 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     7741 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     7801 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7861 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7921 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7981 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     8041 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     8101 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     8161 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     8221 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     8281 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     8341 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     8401 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     8461 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     8521 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     8581 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     8641 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     8701 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     8761 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     8821 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     8881 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     8941 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     9001 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     9061 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     9121 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     9181 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     9241 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     9301 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     9361 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     9421 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctg
//
