LOCUS       sequence-191086-        5550 bp    DNA     linear   UNA 14-APR-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 5550)
  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 5550)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-BECN1
FEATURES             Location/Qualifiers
     source          1..5550
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     341..363
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     425..444
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      491..500
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             506..529
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             566..589
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             596..619
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1728..1737
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2068..2087)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2133..2188
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2187..2206)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2549..2565)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2549..2565)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2562..2584)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2573..2589
                     /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(2597..2603,2604..2621,2622..2627))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2642..2663
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2722..2917
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2768..2903
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2923..3057
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2960..2979)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3014..3033
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3125..3142)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3296..3884)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3376..3395)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4055..4846,4847..4915))
                     /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     4678..4697
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4916..5020)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5051..5430
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
       61 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
      121 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
      181 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
      241 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
      301 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
      361 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
      421 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt cctcgatcga
      481 cggtatcgat gccaccatgg ctagctggag ccacccgcag ttcgagaaag gtggaggtgc
      541 ccgaggtgga tcgggaggtg gatcgtggag ccacccgcag ttcgaaaaag gattcgatta
      601 caaggatgac gatgacaagg gtaccatgga ggggtcaaag acaagcaaca attccactat
      661 gcaggtgtct tttgtgtgcc agcggtgtag tcagcctctg aagctggaca catccttcaa
      721 aatcctggat agagtgacca ttcaggagct gacagctcca ctgctgacca cagctcaggc
      781 taaacctggc gagactcagg aggaagagac caatagcggg gaagagcctt ttatcgagac
      841 cccaaggcag gacggagtgt cccggagatt cattccccct gcccgcatga tgtctactga
      901 gagtgctaat agcttcaccc tgatcggaga ggcttccgat ggagggacca tggaaaacct
      961 gtctaggcgc ctgaaggtga caggggacct gttcgatatt atgtctggac agactgacgt
     1021 ggatcaccca ctgtgcgaag agtgtactga caccctgctg gaccagctgg atacacagct
     1081 gaacgtgact gagaatgaat gccagaacta taagcggtgt ctggagatcc tggaacagat
     1141 gaatgaggac gatagcgaac agctgcagat ggagctgaaa gaactggccc tggaagagga
     1201 acggctgatc caggagctgg aagacgtgga gaagaataga aaaattgtgg ccgagaacct
     1261 ggaaaaggtg caggccgagg ctgaaaggct ggatcaggag gaagctcagt accagaggga
     1321 gtattctgag ttcaagcgcc agcagctgga gctggacgat gaactgaaaa gtgtggagaa
     1381 ccagatgcgc tacgcccaga ctcagctgga taaactgaag aaaaccaacg tgttcaatgc
     1441 tacatttcac atctggcata gcggacagtt cggcaccatt aacaatttta ggctgggaag
     1501 gctgccatcc gtgcccgtgg agtggaacga aatcaatgcc gcttggggac agacagtgct
     1561 gctgctgcac gctctggcta ataagatggg actgaaattc cagcggtaca gactggtgcc
     1621 atatggcaac catagctacc tggagtccct gactgacaag tcaaaagaac tgcccctgta
     1681 ttgcagcgga ggcctgcggt tcttttggga caataagttc gatcatgcca tggtggcttt
     1741 tctggattgc gtgcagcagt tcaaggagga agtggagaaa ggggaaaccc ggttttgtct
     1801 gccctataga atggacgtgg agaagggaaa aatcgaagat accgggggaa gtggcgggtc
     1861 ttacagtatt aagacacagt ttaactcaga ggaacagtgg acaaaggccc tgaaattcat
     1921 gctgactaat ctgaagtggg gcctggcttg ggtgagctcc cagttctaca acaaatgact
     1981 cgaggaattc actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa
     2041 tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat
     2101 catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat
     2161 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     2221 aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc
     2281 catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca
     2341 ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt
     2401 gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt
     2461 cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga gatccctcga
     2521 cctgcagccc aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2581 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     2641 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2701 acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc cgcccctaac
     2761 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     2821 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     2881 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct
     2941 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3001 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatccgc
     3061 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3121 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3181 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3241 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3301 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3361 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3421 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3481 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3541 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3601 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3661 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3721 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3781 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3841 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3901 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3961 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4021 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4081 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4141 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4201 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4261 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4321 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4381 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4441 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4501 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4561 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4621 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4681 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4741 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4801 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4861 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4921 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4981 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5041 cacctgggtc gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     5101 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     5161 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     5221 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     5281 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     5341 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     5401 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact
     5461 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     5521 tgtgcagcga tgggggcggg gggggggggg
//
