LOCUS       sequence-337534-        8904 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the purifcation of human PI3KC3 complex .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8904)
  AUTHORS   Chang C, Shi X, Jensen LE, Yokom AL, Fracchiolla D, Martens S, 
            Hurley JH
  TITLE     Reconstitution of cargo-induced LC3 lipidation in mammalian 
            selective autophagy.
  JOURNAL   Sci Adv. 2021 Apr 23;7(17). pii: 7/17/eabg4922. doi: 
            10.1126/sciadv.abg4922. Print 2021 Apr.
  PUBMED    33893090
REFERENCE   2  (bases 1 to 8904)
  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: "Sci Adv."; 
            date: "2021-04-23"; volume: "7(17). pii: 7/17/eabg4922. doi"; pages:
            " 10.1126/sciadv.abg4922. Print 2021 Apr"
COMMENT     Sequence Label: VPS15
FEATURES             Location/Qualifiers
     source          1..8904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..663
                     /label=chicken beta-actin promoter
     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"
     primer_bind     complement(5926..5945)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5991..6046
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6045..6064)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6407..6423)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6407..6423)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6420..6442)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6431..6447
                     /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(6455..6461,6462..6479,6480..6485))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6500..6521
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6580..6775
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6626..6761
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6781..6915
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6818..6837)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6872..6891
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6983..7000)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7154..7742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7234..7253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7913..8704,8705..8773))
                     /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     8536..8555
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8774..8878)
                     /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 gcgccrggsg gggsggggsg gggsgrgggg
      541 sggggsgggg sgaggcggag aggtgcggcg 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 aggtgggggt gccgggcggg
     1261 gcggggccgc ctcgggccgg ggagggctcg ggggaggggc 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 attcctcgat
     1741 cgacggtatc gaagccacca tgggaaatca gctggccggc atcgctccta gccagattct
     1801 gtccgtggaa tcttacttca gtgacatcca tgattttgag tatgacaaat cactgggaag
     1861 cacccggttc tttaaggtgg cccgggctaa acacagagaa ggcctggtgg tggtgaaggt
     1921 gttcgctatc caggatccca ctctgcctct gaccagttat aaacaggagc tggaggaact
     1981 gaagattaga ctgaactcag cccagaattg cctgccattc cagaaagcta gcgagaaggc
     2041 ctccgaaaaa gccgctatgc tgtttaggca gtacgtgcgc gacaacctgt atgatcggat
     2101 ttccacaaga cccttcctga acaatatcga gaagcggtgg atcgcctttc agattctgac
     2161 tgctgtggac caggcccata aatctggagt gagacacggc gatatcaaga ctgagaacgt
     2221 gatggtgacc agctggaatt gggtgctgct gacagacttc gcttccttta agccaactta
     2281 cctgcccgaa gacaaccctg ccgatttcaa ttatttcttt gacactagcc ggagaaggac
     2341 ctgctacatc gctcctgagc ggttcgtgga tggcgggatg tttgccaccg agctggaata
     2401 catgagagac ccatcaacac ccctggtgga tctgaacagc aatcagagga ggagaggaga
     2461 gctgaagagg gctatggaca tcttttctgc tgggtgtgtg attgccgagc tgttcaccga
     2521 aggagtgcct ctgtttgatc tgagtcagct gctggcctat aggaacggac acttctttcc
     2581 agagcaggtg ctgaataaga tcgaggatca tagcattcgc gaactggtga cacagatgat
     2641 ccacagggaa ccagacaaac gcctggaggc tgaagattac ctgaagcagc agagaggcaa
     2701 cgccttccca gagatcttct acacatttct gcagccctat atggcccagt tcgctaaaga
     2761 gacttttctg agcgccgacg aacggatcct ggtcatcaga aaggatctgg gaaacatcat
     2821 tcataatctg tgcggccacg acctgcccga aaaagccgag ggcgaaccta aggagaatgg
     2881 gctggtcatc ctggtgtctg tgattacaag ttgcctgcag actctgaagt attgtgatag
     2941 caaactggcc gctctggaac tgatcctgca tctggctcca aggctgtccg tggagatcct
     3001 gctggacaga attacccctt acctgctgca cttctccaac gattctgtgc cacgggtgag
     3061 agccgaggct ctgaggacac tgactaaagt gctggccctg gtgaaggaag tgcctcgcaa
     3121 cgacatcaat atctaccccg agtatatcct gcctggcatt gcccacctgg ctcaggacga
     3181 tgccactatt gtgaggctgg cctatgctga gaatatcgcc ctgctggctg aaaccgccct
     3241 gcgctttctg gagctggtgc agctgaagaa cctgaatatg gaaaacgacc ccaacaatga
     3301 ggaaatcgat gaggtgaccc atcctaacgg gaattacgac acagagctgc aggccctgca
     3361 cgaaatggtg cagcagaaag tggtgaccct gctgtccgat ccagaaaaca ttgtgaagca
     3421 gactctgatg gagaatggga tcaccaggct gtgtgtgttc tttggacgcc agaaagctaa
     3481 cgacgtgctg ctgtcacata tgatcacatt cctgaacgac aagaatgatt ggcacctgcg
     3541 gggcgccttc tttgatagca tcgtgggagt ggccgcttat gtgggctggc agagctcctc
     3601 tattctgaag cctctgctgc agcaggggct gtctgacgcc gaggaatttg tgatcgtgaa
     3661 agctctgtat gccctgacct gcatgtgtca gctgggactg ctgcagaagc cacatgtgta
     3721 cgagttcgct agcgatatcg ccccatttct gtgccacccc aatctgtgga ttcggtatgg
     3781 ggccgtggga ttcatcactg tggtggctag acagattagc accgccgacg tgtactgtaa
     3841 actgatgcca tacctggatc cctatatcac acagcccatc attcagattg aacggaaact
     3901 ggtgctgctg tccgtgctga aggagcctgt gtctaggagt attttcgact acgccctgcg
     3961 cagcaaggat atcacttccc tgtttagaca cctgcatatg aggcagaaga aacgcaacgg
     4021 atccctgcct gactgcccac ctccagagga tccagccatc gctcagctgc tgaagaaact
     4081 gctgtctcag gggatgaccg aggaagagga agacaagctg ctggccctga aagatttcat
     4141 gatgaagtct aacaaggcca aagctaatat cgtggaccag agtcacctgc atgatagttc
     4201 acagaagggc gtgatcgacc tggccgctct gggcattaca gggaggcagg tggatctggt
     4261 gaagactaaa caggagcccg acgataaaag ggcccgcaag cacgtgaaac aggacagcaa
     4321 cgtgaatgag gaatggaagt caatgtttgg aagcctggac ccccctaata tgcctcaggc
     4381 tctgccaaaa ggcagcgatc aggaagtgat ccagaccggg aagccaccaa gatcagagag
     4441 ctccgccggc atttgtgtgc ccctgagcac atctagtcag gtgcctgagg tgaccacagt
     4501 gcagaacaag aaacctgtga tcccagtgct gtcaagcacc attctgccaa gcacatacca
     4561 gatcaggatt actacctgca aaaccgaact gcagcagctg atccagcaga agcgggagca
     4621 gtgtaacgct gaaagaattg ccaagcagat gatggagaat gccgagtggg aaagtaaacc
     4681 tccaccacct ggatggaggc caaagggact gctggtggct cacctgcatg aacacaaaag
     4741 cgccgtgaat cggatcagag tgtccgacga gcactctctg ttcgccacat gctccaacga
     4801 tggaactgtg aagatttgga attctcagaa gatggagggc aaaacaacta ccacaaggtc
     4861 tatcctgacc tatagtcgga ttggaggcag agtgaagacc ctgacatttt gtcagggcag
     4921 ccattacctg gccatcgctt ccgacaacgg ggctgtgcag ctgctgggaa ttgaagcctc
     4981 aaagctgccc aaaagcccta agatccaccc cctgcagtcc aggattctgg accagaagga
     5041 ggatggctgc gtggtggaca tgcaccattt caacagtggg gcccagtcag tgctggccta
     5101 cgctacagtg aatgggtctc tggtgggatg ggatctgcgc tcctctagta acgcttggac
     5161 tctgaagcat gacctgaaat ccggcctgat cacctctttc gccgtggata ttcaccagtg
     5221 ctggctgtgt atcggcacat caagcgggac tatggcttgc tgggacatgc gctttcagct
     5281 gccaatctcc tctcattgtc acccaagcag ggctaggatt aggaggctgt ctatgcaccc
     5341 tctgtatcag agttgggtca tcgccgctgt gcaggggaac aatgaagtgt ccatgtggga
     5401 catggagacc ggagatcgga gattcacact gtgggctagt tcagccccac ccctgtctga
     5461 gctgcagcct agtccacatt cagtgcacgg gatctactgt tcccctgctg atggaaatcc
     5521 aatcctgctg accgccggct ctgacatgaa aattaggttt tgggatctgg cttacccaga
     5581 gcgcagttat gtggtggccg gctcaaccag ctccccctca gtgagctact atcggaagat
     5641 cattgagggc acagaagtgg tgcaggaaat ccagaacaag cagaaagtgg gaccaagtga
     5701 cgatactcca aggaggggac ctgagtcact gccagtggga caccatgaca tcattaccga
     5761 tgtggctaca ttccagacta cccagggctt catcgtgacc gcctccagag acggcatcgt
     5821 gaaagtgtgg aagtgactcg aggaattcac tcctcaggtg caggctgcct atcagaaggt
     5881 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc
     5941 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     6001 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     6061 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     6121 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa
     6181 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     6241 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     6301 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     6361 cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc
     6421 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6481 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6541 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac
     6601 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     6661 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     6721 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     6781 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6841 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6901 tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6961 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     7021 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     7081 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     7141 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     7201 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7261 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7321 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7381 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7441 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7501 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7561 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7621 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7681 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7741 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7801 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7861 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7921 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7981 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     8041 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     8101 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     8161 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     8221 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     8281 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8341 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8401 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8461 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8521 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8581 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8641 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8701 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8761 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8821 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8881 catttccccg aaaagtgcca cctg
//
