LOCUS       sequence-193663-        6848 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 6848)
  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 6848)
  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-VPS34
FEATURES             Location/Qualifiers
     source          1..6848
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     331..353
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     415..434
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      481..490
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             496..519
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             556..579
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             586..609
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3369..3388)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3434..3489
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3488..3507)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3850..3866)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3850..3866)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3863..3885)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3874..3890
                     /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(3898..3904,3905..3922,3923..3928))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3943..3964
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4023..4218
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4069..4204
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4224..4358
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4261..4280)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4315..4334
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4426..4443)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4597..5185)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4677..4696)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5356..6147,6148..6216))
                     /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     5979..5998
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6217..6321)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6352..6731
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
       61 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
      121 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
      181 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
      241 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
      301 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
      361 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
      421 tgctggttat tgtgctgtct catcattttg gcaaagaatt cctcgatcga cggtatcgat
      481 gccaccatgg ctagctggag ccacccgcag ttcgagaaag gtggaggtgc ccgaggtgga
      541 tcgggaggtg gatcgtggag ccacccgcag ttcgaaaaag gattcgatta caaggatgac
      601 gatgacaagg gtaccatggg cgaggctgaa aagttccatt acatctattc ctgcgacctg
      661 gatatcaacg tgcagctgaa gattggatcc ctggagggca aacgggaaca gaaatcttat
      721 aaggccgtgc tggaggaccc catgctgaag tttagcggac tgtatcagga aacatgctcc
      781 gatctgtacg tgacttgtca ggtgttcgct gagggaaagc cactggctct gcctgtgagg
      841 acctcctaca aagccttttc tacaagatgg aactggaatg aatggctgaa actgcctgtg
      901 aagtatcctg acctgccaag aaacgcccag gtggctctga ccatctggga tgtgtacgga
      961 cctggaaagg ctgtgccagt gggagggacc acagtgtccc tgttcgggaa gtacggaatg
     1021 tttcggcagg ggatgcacga cctgaaagtg tggcccaatg tggaggccga tggctctgaa
     1081 ccaactaaga cccccgggag aacaagctcc actctgagtg aggaccagat gtcaaggctg
     1141 gccaaactga ctaaggctca ccgccaggga catatggtga aggtggactg gctggatcgg
     1201 ctgaccttca gagagatcga aatgattaac gagagcgaaa aacggtctag taatttcatg
     1261 tatctgatgg tggagtttag atgcgtgaaa tgtgacgata aggaatacgg aatcgtgtac
     1321 tatgagaagg acggcgatga atcaagcccc attctgacca gctttgagct ggtgaaagtg
     1381 ccagaccccc agatgagtat ggagaacctg gtggaatcaa aacaccataa gctggcccgg
     1441 tccctgagat ctggccctag tgaccatgat ctgaagccaa acgccgctac aagggaccag
     1501 ctgaatatca ttgtgagcta tccccctact aaacagctga cctacgagga acaggatctg
     1561 gtgtggaagt tccgctacta tctgactaac caggagaaag ccctgaccaa atttctgaag
     1621 tgcgtgaatt gggacctgcc tcaggaggcc aagcaggctc tggaactgct gggcaaatgg
     1681 aagccaatgg acgtggagga tagcctggaa ctgctgtcct ctcactatac caatccaaca
     1741 gtgcggagat acgctgtggc taggctgagg caggctgacg atgaggatct gctgatgtat
     1801 ctgctgcagc tggtgcaggc cctgaagtac gagaacttcg acgatatcaa aaatgggctg
     1861 gaacccacca agaaagacag ccagagttca gtgtccgaga acgtgtcaaa tagcggaatc
     1921 aacagcgctg aaattgatag ctcccagatc attacctcac ctctgccaag cgtgtctagt
     1981 ccaccccctg cctccaaaac aaaggaggtg cctgacggcg agaatctgga acaggatctg
     2041 tgcaccttcc tgatctcacg ggcctgtaag aacagcacac tggctaatta cctgtattgg
     2101 tacgtgattg tggagtgtga agaccaggat acacagcaga gagacccaaa gactcatgag
     2161 atgtacctga acgtgatgag gcgcttcagc caggccctgc tgaaagggga caagtccgtg
     2221 agggtgatgc gctctctgct ggccgctcag cagacctttg tggatcggct ggtgcacctg
     2281 atgaaggctg tgcagaggga gtctgggaac cgcaagaaaa agaatgaaag actgcaggcc
     2341 ctgctgggag acaacgagaa aatgaatctg agtgatgtgg agctgatccc cctgcctctg
     2401 gaaccccagg tgaagattag aggaatcatt cctgagacag ccactctgtt caaatccgcc
     2461 ctgatgccag ctcagctgtt ctttaaaaca gaagacggcg gcaagtatcc cgtgatcttt
     2521 aaacatggcg acgatctgag gcaggatcag ctgattctgc agatcatttc tctgatggac
     2581 aaactgctgc gcaaggagaa cctggatctg aaactgacac cctacaaggt gctggctacc
     2641 agtacaaagc acggcttcat gcagtttatc cagtcagtgc ctgtggccga ggtgctggac
     2701 actgaaggga gcattcagaa tttctttagg aagtatgccc cctctgagaa cggccctaat
     2761 gggatcagtg ctgaagtgat ggatacttac gtgaaaagct gcgccggcta ttgtgtgatc
     2821 acctacattc tgggagtggg cgacaggcat ctggataacc tgctgctgac taaaaccggg
     2881 aagctgttcc acatcgactt tgggtatatt ctgggaaggg atccaaagcc actgccacca
     2941 cctatgaaac tgaataagga gatggtggaa ggcatggggg gaacacagtc cgagcagtat
     3001 caggaattcc gcaagcagtg ttacactgcc tttctgcacc tgcggagata ctcaaacctg
     3061 atcctgaatc tgttcagcct gatggtggac gccaatatcc cagatattgc tctggagccc
     3121 gacaagaccg tgaaaaaggt gcaggataaa tttaggctgg acctgtctga tgaggaagcc
     3181 gtgcactata tgcagtctct gatcgacgag agtgtgcatg ctctgttcgc cgctgtggtg
     3241 gaacagattc acaagtttgc ccagtactgg cgcaaatgac tcgaggaatt cactcctcag
     3301 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     3361 cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     3421 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     3481 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     3541 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     3601 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     3661 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     3721 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     3781 ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc
     3841 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3901 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3961 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat
     4021 ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4081 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4141 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4201 ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa
     4261 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4321 tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca
     4381 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4441 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4501 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4561 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4621 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4681 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4741 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4801 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4861 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4921 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4981 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5041 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5101 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5161 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5221 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5281 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5341 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5401 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5461 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5521 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5581 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5641 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5701 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5761 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5821 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5881 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5941 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6001 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6061 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6121 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6181 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6241 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6301 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt cgacattgat
     6361 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     6421 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     6481 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     6541 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     6601 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     6661 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     6721 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
     6781 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg
     6841 gggggggg
//
