LOCUS       sequence-332451-        8903 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 8903)
  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 8903)
  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..8903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1753..1762
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5925..5944)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5990..6045
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6044..6063)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6406..6422)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6406..6422)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6419..6441)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6430..6446
                     /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(6454..6460,6461..6478,6479..6484))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6499..6520
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6579..6774
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6625..6760
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6780..6914
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6817..6836)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6871..6890
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6982..6999)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7153..7741)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7233..7252)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7912..8703,8704..8772))
                     /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     8535..8554
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8773..8877)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgcgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcctcgatc
     1741 gacggtatcg atgccaccat gggaaatcag ctggccggca tcgctcctag ccagattctg
     1801 tccgtggaat cttacttcag tgacatccat gattttgagt atgacaaatc actgggaagc
     1861 acccggttct ttaaggtggc ccgggctaaa cacagagaag gcctggtggt ggtgaaggtg
     1921 ttcgctatcc aggatcccac tctgcctctg accagttata aacaggagct ggaggaactg
     1981 aagattagac tgaactcagc ccagaattgc ctgccattcc agaaagctag cgagaaggcc
     2041 tccgaaaaag ccgctatgct gtttaggcag tacgtgcgcg acaacctgta tgatcggatt
     2101 tccacaagac ccttcctgaa caatatcgag aagcggtgga tcgcctttca gattctgact
     2161 gctgtggacc aggcccataa atctggagtg agacacggcg atatcaagac tgagaacgtg
     2221 atggtgacca gctggaattg ggtgctgctg acagacttcg cttcctttaa gccaacttac
     2281 ctgcccgaag acaaccctgc cgatttcaat tatttctttg acactagccg gagaaggacc
     2341 tgctacatcg ctcctgagcg gttcgtggat ggcgggatgt ttgccaccga gctggaatac
     2401 atgagagacc catcaacacc cctggtggat ctgaacagca atcagaggag gagaggagag
     2461 ctgaagaggg ctatggacat cttttctgct gggtgtgtga ttgccgagct gttcaccgaa
     2521 ggagtgcctc tgtttgatct gagtcagctg ctggcctata ggaacggaca cttctttcca
     2581 gagcaggtgc tgaataagat cgaggatcat agcattcgcg aactggtgac acagatgatc
     2641 cacagggaac cagacaaacg cctggaggct gaagattacc tgaagcagca gagaggcaac
     2701 gccttcccag agatcttcta cacatttctg cagccctata tggcccagtt cgctaaagag
     2761 acttttctga gcgccgacga acggatcctg gtcatcagaa aggatctggg aaacatcatt
     2821 cataatctgt gcggccacga cctgcccgaa aaagccgagg gcgaacctaa ggagaatggg
     2881 ctggtcatcc tggtgtctgt gattacaagt tgcctgcaga ctctgaagta ttgtgatagc
     2941 aaactggccg ctctggaact gatcctgcat ctggctccaa ggctgtccgt ggagatcctg
     3001 ctggacagaa ttacccctta cctgctgcac ttctccaacg attctgtgcc acgggtgaga
     3061 gccgaggctc tgaggacact gactaaagtg ctggccctgg tgaaggaagt gcctcgcaac
     3121 gacatcaata tctaccccga gtatatcctg cctggcattg cccacctggc tcaggacgat
     3181 gccactattg tgaggctggc ctatgctgag aatatcgccc tgctggctga aaccgccctg
     3241 cgctttctgg agctggtgca gctgaagaac ctgaatatgg aaaacgaccc caacaatgag
     3301 gaaatcgatg aggtgaccca tcctaacggg aattacgaca cagagctgca ggccctgcac
     3361 gaaatggtgc agcagaaagt ggtgaccctg ctgtccgatc cagaaaacat tgtgaagcag
     3421 actctgatgg agaatgggat caccaggctg tgtgtgttct ttggacgcca gaaagctaac
     3481 gacgtgctgc tgtcacatat gatcacattc ctgaacgaca agaatgattg gcacctgcgg
     3541 ggcgccttct ttgatagcat cgtgggagtg gccgcttatg tgggctggca gagctcctct
     3601 attctgaagc ctctgctgca gcaggggctg tctgacgccg aggaatttgt gatcgtgaaa
     3661 gctctgtatg ccctgacctg catgtgtcag ctgggactgc tgcagaagcc acatgtgtac
     3721 gagttcgcta gcgatatcgc cccatttctg tgccacccca atctgtggat tcggtatggg
     3781 gccgtgggat tcatcactgt ggtggctaga cagattagca ccgccgacgt gtactgtaaa
     3841 ctgatgccat acctggatcc ctatatcaca cagcccatca ttcagattga acggaaactg
     3901 gtgctgctgt ccgtgctgaa ggagcctgtg tctaggagta ttttcgacta cgccctgcgc
     3961 agcaaggata tcacttccct gtttagacac ctgcatatga ggcagaagaa acgcaacgga
     4021 tccctgcctg actgcccacc tccagaggat ccagccatcg ctcagctgct gaagaaactg
     4081 ctgtctcagg ggatgaccga ggaagaggaa gacaagctgc tggccctgaa agatttcatg
     4141 atgaagtcta acaaggccaa agctaatatc gtggaccaga gtcacctgca tgatagttca
     4201 cagaagggcg tgatcgacct ggccgctctg ggcattacag ggaggcaggt ggatctggtg
     4261 aagactaaac aggagcccga cgataaaagg gcccgcaagc acgtgaaaca ggacagcaac
     4321 gtgaatgagg aatggaagtc aatgtttgga agcctggacc cccctaatat gcctcaggct
     4381 ctgccaaaag gcagcgatca ggaagtgatc cagaccggga agccaccaag atcagagagc
     4441 tccgccggca tttgtgtgcc cctgagcaca tctagtcagg tgcctgaggt gaccacagtg
     4501 cagaacaaga aacctgtgat cccagtgctg tcaagcacca ttctgccaag cacataccag
     4561 atcaggatta ctacctgcaa aaccgaactg cagcagctga tccagcagaa gcgggagcag
     4621 tgtaacgctg aaagaattgc caagcagatg atggagaatg ccgagtggga aagtaaacct
     4681 ccaccacctg gatggaggcc aaagggactg ctggtggctc acctgcatga acacaaaagc
     4741 gccgtgaatc ggatcagagt gtccgacgag cactctctgt tcgccacatg ctccaacgat
     4801 ggaactgtga agatttggaa ttctcagaag atggagggca aaacaactac cacaaggtct
     4861 atcctgacct atagtcggat tggaggcaga gtgaagaccc tgacattttg tcagggcagc
     4921 cattacctgg ccatcgcttc cgacaacggg gctgtgcagc tgctgggaat tgaagcctca
     4981 aagctgccca aaagccctaa gatccacccc ctgcagtcca ggattctgga ccagaaggag
     5041 gatggctgcg tggtggacat gcaccatttc aacagtgggg cccagtcagt gctggcctac
     5101 gctacagtga atgggtctct ggtgggatgg gatctgcgct cctctagtaa cgcttggact
     5161 ctgaagcatg acctgaaatc cggcctgatc acctctttcg ccgtggatat tcaccagtgc
     5221 tggctgtgta tcggcacatc aagcgggact atggcttgct gggacatgcg ctttcagctg
     5281 ccaatctcct ctcattgtca cccaagcagg gctaggatta ggaggctgtc tatgcaccct
     5341 ctgtatcaga gttgggtcat cgccgctgtg caggggaaca atgaagtgtc catgtgggac
     5401 atggagaccg gagatcggag attcacactg tgggctagtt cagccccacc cctgtctgag
     5461 ctgcagccta gtccacattc agtgcacggg atctactgtt cccctgctga tggaaatcca
     5521 atcctgctga ccgccggctc tgacatgaaa attaggtttt gggatctggc ttacccagag
     5581 cgcagttatg tggtggccgg ctcaaccagc tccccctcag tgagctacta tcggaagatc
     5641 attgagggca cagaagtggt gcaggaaatc cagaacaagc agaaagtggg accaagtgac
     5701 gatactccaa ggaggggacc tgagtcactg ccagtgggac accatgacat cattaccgat
     5761 gtggctacat tccagactac ccagggcttc atcgtgaccg cctccagaga cggcatcgtg
     5821 aaagtgtgga agtgactcga ggaattcact cctcaggtgc aggctgccta tcagaaggtg
     5881 gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc
     5941 aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa
     6001 tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat
     6061 gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat
     6121 gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa
     6181 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
     6241 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
     6301 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
     6361 ttatggagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc
     6421 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6481 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6541 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc
     6601 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     6661 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     6721 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta
     6781 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6841 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6901 atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6961 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     7021 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7081 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7141 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7201 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7261 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7321 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     7381 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7441 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7501 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7561 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     7621 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7681 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7741 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7801 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7861 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7921 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7981 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     8041 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     8101 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     8161 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     8221 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8281 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8341 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8401 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8461 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8521 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8581 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8641 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8701 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8761 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8821 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8881 atttccccga aaagtgccac ctg
//
