LOCUS       sequence-191081-        8267 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Component for mammalian expression of recombinant PI3KC3-C1 complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8267)
  AUTHORS   Baskaran S, Carlson LA, Stjepanovic G, Young LN, Kim DJ, Grob P, 
            Stanley RE, Nogales E, Hurley JH
  TITLE     Architecture and dynamics of the autophagic phosphatidylinositol 
            3-kinase complex.
  JOURNAL   Elife. 2014 Dec 9;3. doi: 10.7554/eLife.05115.
  PUBMED    25490155
REFERENCE   2  (bases 1 to 8267)
  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: "Elife."; 
            date: "2014-12-9"; volume: "3. doi"; pages: " 10.7554/eLife.05115"
COMMENT     Sequence Label: pCAG-OSF-VPS15
FEATURES             Location/Qualifiers
     source          1..8267
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     336..358
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     420..439
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      486..495
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             501..524
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             561..584
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             591..614
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4787..4806)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4852..4907
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4906..4925)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5268..5284)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5268..5284)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5281..5303)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5292..5308
                     /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(5316..5322,5323..5340,5341..5346))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5361..5382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5441..5636
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5487..5622
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5642..5776
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5679..5698)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5733..5752
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5844..5861)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6015..6603)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6095..6114)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6774..7565,7566..7634))
                     /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     7397..7416
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7635..7739)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7770..8149
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg
       61 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg
      121 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc
      181 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
      241 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
      301 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
      361 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
      421 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattcctcg atcgacggta
      481 tcgatgccac catggctagc tggagccacc cgcagttcga gaaaggtgga ggtgcccgag
      541 gtggatcggg aggtggatcg tggagccacc cgcagttcga aaaaggattc gattacaagg
      601 atgacgatga caagggtacc atgggaaatc agctggccgg catcgctcct agccagattc
      661 tgtccgtgga atcttacttc agtgacatcc atgattttga gtatgacaaa tcactgggaa
      721 gcacccggtt ctttaaggtg gcccgggcta aacacagaga aggcctggtg gtggtgaagg
      781 tgttcgctat ccaggatccc actctgcctc tgaccagtta taaacaggag ctggaggaac
      841 tgaagattag actgaactca gcccagaatt gcctgccatt ccagaaagct agcgagaagg
      901 cctccgaaaa agccgctatg ctgtttaggc agtacgtgcg cgacaacctg tatgatcgga
      961 tttccacaag acccttcctg aacaatatcg agaagcggtg gatcgccttt cagattctga
     1021 ctgctgtgga ccaggcccat aaatctggag tgagacacgg cgatatcaag actgagaacg
     1081 tgatggtgac cagctggaat tgggtgctgc tgacagactt cgcttccttt aagccaactt
     1141 acctgcccga agacaaccct gccgatttca attatttctt tgacactagc cggagaagga
     1201 cctgctacat cgctcctgag cggttcgtgg atggcgggat gtttgccacc gagctggaat
     1261 acatgagaga cccatcaaca cccctggtgg atctgaacag caatcagagg aggagaggag
     1321 agctgaagag ggctatggac atcttttctg ctgggtgtgt gattgccgag ctgttcaccg
     1381 aaggagtgcc tctgtttgat ctgagtcagc tgctggccta taggaacgga cacttctttc
     1441 cagagcaggt gctgaataag atcgaggatc atagcattcg cgaactggtg acacagatga
     1501 tccacaggga accagacaaa cgcctggagg ctgaagatta cctgaagcag cagagaggca
     1561 acgccttccc agagatcttc tacacatttc tgcagcccta tatggcccag ttcgctaaag
     1621 agacttttct gagcgccgac gaacggatcc tggtcatcag aaaggatctg ggaaacatca
     1681 ttcataatct gtgcggccac gacctgcccg aaaaagccga gggcgaacct aaggagaatg
     1741 ggctggtcat cctggtgtct gtgattacaa gttgcctgca gactctgaag tattgtgata
     1801 gcaaactggc cgctctggaa ctgatcctgc atctggctcc aaggctgtcc gtggagatcc
     1861 tgctggacag aattacccct tacctgctgc acttctccaa cgattctgtg ccacgggtga
     1921 gagccgaggc tctgaggaca ctgactaaag tgctggccct ggtgaaggaa gtgcctcgca
     1981 acgacatcaa tatctacccc gagtatatcc tgcctggcat tgcccacctg gctcaggacg
     2041 atgccactat tgtgaggctg gcctatgctg agaatatcgc cctgctggct gaaaccgccc
     2101 tgcgctttct ggagctggtg cagctgaaga acctgaatat ggaaaacgac cccaacaatg
     2161 aggaaatcga tgaggtgacc catcctaacg ggaattacga cacagagctg caggccctgc
     2221 acgaaatggt gcagcagaaa gtggtgaccc tgctgtccga tccagaaaac attgtgaagc
     2281 agactctgat ggagaatggg atcaccaggc tgtgtgtgtt ctttggacgc cagaaagcta
     2341 acgacgtgct gctgtcacat atgatcacat tcctgaacga caagaatgat tggcacctgc
     2401 ggggcgcctt ctttgatagc atcgtgggag tggccgctta tgtgggctgg cagagctcct
     2461 ctattctgaa gcctctgctg cagcaggggc tgtctgacgc cgaggaattt gtgatcgtga
     2521 aagctctgta tgccctgacc tgcatgtgtc agctgggact gctgcagaag ccacatgtgt
     2581 acgagttcgc tagcgatatc gccccatttc tgtgccaccc caatctgtgg attcggtatg
     2641 gggccgtggg attcatcact gtggtggcta gacagattag caccgccgac gtgtactgta
     2701 aactgatgcc atacctggat ccctatatca cacagcccat cattcagatt gaacggaaac
     2761 tggtgctgct gtccgtgctg aaggagcctg tgtctaggag tattttcgac tacgccctgc
     2821 gcagcaagga tatcacttcc ctgtttagac acctgcatat gaggcagaag aaacgcaacg
     2881 gatccctgcc tgactgccca cctccagagg atccagccat cgctcagctg ctgaagaaac
     2941 tgctgtctca ggggatgacc gaggaagagg aagacaagct gctggccctg aaagatttca
     3001 tgatgaagtc taacaaggcc aaagctaata tcgtggacca gagtcacctg catgatagtt
     3061 cacagaaggg cgtgatcgac ctggccgctc tgggcattac agggaggcag gtggatctgg
     3121 tgaagactaa acaggagccc gacgataaaa gggcccgcaa gcacgtgaaa caggacagca
     3181 acgtgaatga ggaatggaag tcaatgtttg gaagcctgga cccccctaat atgcctcagg
     3241 ctctgccaaa aggcagcgat caggaagtga tccagaccgg gaagccacca agatcagaga
     3301 gctccgccgg catttgtgtg cccctgagca catctagtca ggtgcctgag gtgaccacag
     3361 tgcagaacaa gaaacctgtg atcccagtgc tgtcaagcac cattctgcca agcacatacc
     3421 agatcaggat tactacctgc aaaaccgaac tgcagcagct gatccagcag aagcgggagc
     3481 agtgtaacgc tgaaagaatt gccaagcaga tgatggagaa tgccgagtgg gaaagtaaac
     3541 ctccaccacc tggatggagg ccaaagggac tgctggtggc tcacctgcat gaacacaaaa
     3601 gcgccgtgaa tcggatcaga gtgtccgacg agcactctct gttcgccaca tgctccaacg
     3661 atggaactgt gaagatttgg aattctcaga agatggaggg caaaacaact accacaaggt
     3721 ctatcctgac ctatagtcgg attggaggca gagtgaagac cctgacattt tgtcagggca
     3781 gccattacct ggccatcgct tccgacaacg gggctgtgca gctgctggga attgaagcct
     3841 caaagctgcc caaaagccct aagatccacc ccctgcagtc caggattctg gaccagaagg
     3901 aggatggctg cgtggtggac atgcaccatt tcaacagtgg ggcccagtca gtgctggcct
     3961 acgctacagt gaatgggtct ctggtgggat gggatctgcg ctcctctagt aacgcttgga
     4021 ctctgaagca tgacctgaaa tccggcctga tcacctcttt cgccgtggat attcaccagt
     4081 gctggctgtg tatcggcaca tcaagcggga ctatggcttg ctgggacatg cgctttcagc
     4141 tgccaatctc ctctcattgt cacccaagca gggctaggat taggaggctg tctatgcacc
     4201 ctctgtatca gagttgggtc atcgccgctg tgcaggggaa caatgaagtg tccatgtggg
     4261 acatggagac cggagatcgg agattcacac tgtgggctag ttcagcccca cccctgtctg
     4321 agctgcagcc tagtccacat tcagtgcacg ggatctactg ttcccctgct gatggaaatc
     4381 caatcctgct gaccgccggc tctgacatga aaattaggtt ttgggatctg gcttacccag
     4441 agcgcagtta tgtggtggcc ggctcaacca gctccccctc agtgagctac tatcggaaga
     4501 tcattgaggg cacagaagtg gtgcaggaaa tccagaacaa gcagaaagtg ggaccaagtg
     4561 acgatactcc aaggagggga cctgagtcac tgccagtggg acaccatgac atcattaccg
     4621 atgtggctac attccagact acccagggct tcatcgtgac cgcctccaga gacggcatcg
     4681 tgaaagtgtg gaagtgactc gaggaattca ctcctcaggt gcaggctgcc tatcagaagg
     4741 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     4801 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     4861 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     4921 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     4981 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     5041 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     5101 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     5161 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     5221 tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt
     5281 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5341 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5401 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa
     5461 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5521 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     5581 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     5641 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5701 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5761 ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5821 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5881 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5941 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6001 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6061 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6121 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6181 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6241 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6301 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6361 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6421 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6481 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6541 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6601 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6661 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6721 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6781 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6841 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6901 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6961 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7021 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7081 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7141 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7201 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7261 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7321 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7381 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7441 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7501 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7561 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7621 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7681 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7741 acatttcccc gaaaagtgcc acctgggtcg acattgatta ttgactagtt attaatagta
     7801 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     7861 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     7921 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     7981 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     8041 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     8101 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
     8161 ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt
     8221 atttattttt taattatttt gtgcagcgat gggggcgggg ggggggg
//
