LOCUS       sequence-400839-        7490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of VPS34.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7490)
  AUTHORS   Chen M, Ren X, Nguyen TN, Khuu G, Cook ASI, Lazarou M, Hurley JH
  TITLE     Structure and activation of the human autophagy-initiating 
            ULK1C:PI3KC3-C1 supercomplex
  JOURNAL   bioRxiv 2023.06.01.543278
REFERENCE   2  (bases 1 to 7490)
  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: "bioRxiv 
            2023.06.01.543278"
COMMENT     Sequence Label: pCAG-VPS34
FEATURES             Location/Qualifiers
     source          1..7490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        373..648
                     /label=chicken beta-actin promoter
     intron          649..1665
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1589..1611
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1673..1692
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1740..1749
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4499..4518)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4564..4619
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4618..4637)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4980..4996)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4980..4996)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4993..5015)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5004..5020
                     /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(5028..5034,5035..5052,5053..5058))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5073..5094
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5153..5348
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5199..5334
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5354..5488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5391..5410)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5445..5464
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5556..5573)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5727..6315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5807..5826)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6486..7277,7278..7346))
                     /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     7109..7128
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7347..7451)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(7482..7490,1..371)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
      421 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg
      481 gggggggggg ggsgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
      541 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
      601 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg
      661 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
      721 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
      781 tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg
      841 ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc
      901 gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc
      961 gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg
     1021 cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg
     1081 tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga
     1141 gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc
     1201 gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg
     1261 gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc
     1321 gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct
     1381 ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg
     1441 cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg
     1501 tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc
     1561 tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct
     1621 agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg
     1681 ctggttattg tgctgtctca tcattttggc aaagaattcc tcgatcgacg gtatcgatag
     1741 ccaccatggg cgaggctgaa aagttccatt acatctattc ctgcgacctg gatatcaacg
     1801 tgcagctgaa gattggatcc ctggagggca aacgggaaca gaaatcttat aaggccgtgc
     1861 tggaggaccc catgctgaag tttagcggac tgtatcagga aacatgctcc gatctgtacg
     1921 tgacttgtca ggtgttcgct gagggaaagc cactggctct gcctgtgagg acctcctaca
     1981 aagccttttc tacaagatgg aactggaatg aatggctgaa actgcctgtg aagtatcctg
     2041 acctgccaag aaacgcccag gtggctctga ccatctggga tgtgtacgga cctggaaagg
     2101 ctgtgccagt gggagggacc acagtgtccc tgttcgggaa gtacggaatg tttcggcagg
     2161 ggatgcacga cctgaaagtg tggcccaatg tggaggccga tggctctgaa ccaactaaga
     2221 cccccgggag aacaagctcc actctgagtg aggaccagat gtcaaggctg gccaaactga
     2281 ctaaggctca ccgccaggga catatggtga aggtggactg gctggatcgg ctgaccttca
     2341 gagagatcga aatgattaac gagagcgaaa aacggtctag taatttcatg tatctgatgg
     2401 tggagtttag atgcgtgaaa tgtgacgata aggaatacgg aatcgtgtac tatgagaagg
     2461 acggcgatga atcaagcccc attctgacca gctttgagct ggtgaaagtg ccagaccccc
     2521 agatgagtat ggagaacctg gtggaatcaa aacaccataa gctggcccgg tccctgagat
     2581 ctggccctag tgaccatgat ctgaagccaa acgccgctac aagggaccag ctgaatatca
     2641 ttgtgagcta tccccctact aaacagctga cctacgagga acaggatctg gtgtggaagt
     2701 tccgctacta tctgactaac caggagaaag ccctgaccaa atttctgaag tgcgtgaatt
     2761 gggacctgcc tcaggaggcc aagcaggctc tggaactgct gggcaaatgg aagccaatgg
     2821 acgtggagga tagcctggaa ctgctgtcct ctcactatac caatccaaca gtgcggagat
     2881 acgctgtggc taggctgagg caggctgacg atgaggatct gctgatgtat ctgctgcagc
     2941 tggtgcaggc cctgaagtac gagaacttcg acgatatcaa aaatgggctg gaacccacca
     3001 agaaagacag ccagagttca gtgtccgaga acgtgtcaaa tagcggaatc aacagcgctg
     3061 aaattgatag ctcccagatc attacctcac ctctgccaag cgtgtctagt ccaccccctg
     3121 cctccaaaac aaaggaggtg cctgacggcg agaatctgga acaggatctg tgcaccttcc
     3181 tgatctcacg ggcctgtaag aacagcacac tggctaatta cctgtattgg tacgtgattg
     3241 tggagtgtga agaccaggat acacagcaga gagacccaaa gactcatgag atgtacctga
     3301 acgtgatgag gcgcttcagc caggccctgc tgaaagggga caagtccgtg agggtgatgc
     3361 gctctctgct ggccgctcag cagacctttg tggatcggct ggtgcacctg atgaaggctg
     3421 tgcagaggga gtctgggaac cgcaagaaaa agaatgaaag actgcaggcc ctgctgggag
     3481 acaacgagaa aatgaatctg agtgatgtgg agctgatccc cctgcctctg gaaccccagg
     3541 tgaagattag aggaatcatt cctgagacag ccactctgtt caaatccgcc ctgatgccag
     3601 ctcagctgtt ctttaaaaca gaagacggcg gcaagtatcc cgtgatcttt aaacatggcg
     3661 acgatctgag gcaggatcag ctgattctgc agatcatttc tctgatggac aaactgctgc
     3721 gcaaggagaa cctggatctg aaactgacac cctacaaggt gctggctacc agtacaaagc
     3781 acggcttcat gcagtttatc cagtcagtgc ctgtggccga ggtgctggac actgaaggga
     3841 gcattcagaa tttctttagg aagtatgccc cctctgagaa cggccctaat gggatcagtg
     3901 ctgaagtgat ggatacttac gtgaaaagct gcgccggcta ttgtgtgatc acctacattc
     3961 tgggagtggg cgacaggcat ctggataacc tgctgctgac taaaaccggg aagctgttcc
     4021 acatcgactt tgggtatatt ctgggaaggg atccaaagcc actgccacca cctatgaaac
     4081 tgaataagga gatggtggaa ggcatggggg gaacacagtc cgagcagtat caggaattcc
     4141 gcaagcagtg ttacactgcc tttctgcacc tgcggagata ctcaaacctg atcctgaatc
     4201 tgttcagcct gatggtggac gccaatatcc cagatattgc tctggagccc gacaagaccg
     4261 tgaaaaaggt gcaggataaa tttaggctgg acctgtctga tgaggaagcc gtgcactata
     4321 tgcagtctct gatcgacgag agtgtgcatg ctctgttcgc cgctgtggtg gaacagattc
     4381 acaagtttgc ccagtactgg cgcaaatgac tcgaggaatt cactcctcag gtgcaggctg
     4441 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
     4501 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     4561 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     4621 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     4681 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     4741 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     4801 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     4861 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     4921 ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg
     4981 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5041 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5101 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     5161 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5221 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5281 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5341 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5401 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5461 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     5521 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5581 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5641 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5701 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5761 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5821 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5881 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5941 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6001 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6061 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6121 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6181 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6241 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6301 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6361 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6421 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6481 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6541 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6601 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6661 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6721 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6781 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6841 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6901 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6961 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7021 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7081 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7141 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7201 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7261 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7321 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7381 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7441 ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt cgacattgat
//
