LOCUS       sequence-191085-        6714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Component for mammalian expression of recombinant PI3KC3-C1 complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6714)
  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 6714)
  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: pLEXm-GST-ATG14
FEATURES             Location/Qualifiers
     source          1..6714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          168..1184
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1108..1130
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1192..1211
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1260..1269
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1266..1922
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MASPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
                     KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIR
                     YGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVV
                     LYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1880..1902
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1950..1970
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3578..3597)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3643..3698
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3697..3716)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4059..4075)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4059..4075)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4072..4094)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4083..4099
                     /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(4107..4113,4114..4131,4132..4137))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4152..4173
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4302..4319)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4473..5061)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4553..4572)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5232..6023,6024..6092))
                     /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     5855..5874
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6093..6197)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6228..6607
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6609..6714,1..167)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc
       61 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
      121 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
      181 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
      241 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
      301 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc
      361 tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg
      421 tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg
      481 cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc
      541 ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt
      601 gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag
      661 ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg
      721 ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg
      781 ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg
      841 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt
      901 tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc
      961 gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt
     1021 cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct
     1081 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     1141 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     1201 tggttattgt gctgtctcat cattttggca aagaattgcg gccgtctcag gccgaattcg
     1261 ccaccatggc atcccctata ctaggttatt ggaaaattaa gggccttgtg caacccactc
     1321 gacttctttt ggaatatctt gaagaaaaat atgaagagca tttgtatgag cgcgatgaag
     1381 gtgataaatg gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat cttccttatt
     1441 atattgatgg tgatgttaaa ttaacacagt ctatggccat catacgttat atagctgaca
     1501 agcacaacat gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag
     1561 cggttttgga tattagatac ggtgtttcga gaattgcata tagtaaagac tttgaaactc
     1621 tcaaagttga ttttcttagc aagctacctg aaatgctgaa aatgttcgaa gatcgtttat
     1681 gtcataaaac atatttaaat ggtgatcatg taacccatcc tgacttcatg ttgtatgacg
     1741 ctcttgatgt tgttttatac atggacccaa tgtgcctgga tgcgttccca aaattagttt
     1801 gttttaaaaa acgtattgaa gctatcccac aaattgataa gtacttgaaa tccagcaagt
     1861 atatagcatg gcctttgcag ggctggcaag ccacgtttgg tggtggcgac catcctccaa
     1921 aatcggatga ttacgatatc ccaacgaccg aaaacctgta ttttcagggt gtcgacggta
     1981 ccatggcttc cccatctgga aagggagcta gggctctgga ggctcctgga tgcggaccac
     2041 ggcccctggc cagagacctg gtggatagcg tggacgatgc tgaggggctg tacgtggccg
     2101 tggaaagatg ccccctgtgt aataccacac ggagaaggct gacatgcgcc aagtgtgtgc
     2161 agtccgggga tttcgtgtat tttgacggac gcgatcggga gagattcatt gacaagaaag
     2221 aaaggctgtc ccgcctgaag tctaaacagg aggaatttca gaaagaggtg ctgaaggcca
     2281 tggagggcaa gtggatcact gatcagctga ggtggaaaat catgtcctgc aagatgcgca
     2341 ttgagcagct gaaacagacc atctgtaagg gaaacgagga aatggagaaa aatagcgaag
     2401 gcctgctgaa gacaaaagaa aagaatcaga agctgtactc ccgggctcag agacaccagg
     2461 agaagaaaga aaaaattcag aggcataacc gcaagctggg cgatctggtg gagaagaaaa
     2521 ccatcgacct gaggtctcac tatgaacgcc tggccaatct gcgccggagc catatcctgg
     2581 agctgacatc cgtgatcttc ccaattgagg aagtgaaaac tggagtgagg gaccccgctg
     2641 atgtgagctc cgagagtgac tcagccatga cctctagtac agtgagtaag ctggctgaag
     2701 ccagaaggac tacctacctg tcaggccgct gggtgtgcga cgatcacaac ggcgatacaa
     2761 gcatctccat tactgggccc tggattagcc tgcctaacaa tggcgactat agtgcctact
     2821 attcatgggt ggaggaaaag aaaacaactc aggggccaga tatggagcag tctaatcccg
     2881 cttacactat cagtgccgct ctgtgctatg ccacccagct ggtgaacatc ctgtctcata
     2941 ttctggacgt gaacctgccc aagaaactgt gcaattcaga gttctgtggc gaaaacctga
     3001 gcaaacagaa gtttactagg gctgtgaaga aactgaacgc caatattctg tacctgtgtt
     3061 ttagccagca cgtgaatctg gatcagctgc agcctctgca taccctgcgc aacctgatgt
     3121 atctggtgtc cccttcaagc gagcacctgg gaaggtctgg accattcgaa gtgagagctg
     3181 atctggagga aagcatggag tttgtggacc ctggagtggc cggcgagtct gacgaaagtg
     3241 gggatgagcg ggtgtctgac gaggaaactg atctgggaac cgactgggaa aacctgccta
     3301 gtccaagatt ctgtgacatc ccatctcaga gtgtggaggt gtcacagagc cagtccacac
     3361 aggcttcccc ccctattgct tcctctagtg ctggaggaat gatctcaagc gccgctgcct
     3421 cagtgaccag ctggtttaag gcctacacag gccataggtg agcggccgcg catgcctcga
     3481 gactagtatc gcgataattc actcctcagg tgcaggctgc ctatcagaag gtggtggctg
     3541 gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt
     3601 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     3661 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     3721 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     3781 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     3841 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     3901 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     3961 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga
     4021 gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     4081 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4141 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4201 cagtcgggaa acctgtcgtg ccagcggatc gatccgctgc attaatgaat cggccaacgc
     4261 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4321 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4381 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4441 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4501 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4561 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4621 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4681 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4741 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4801 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4861 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4921 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4981 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5041 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5101 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5161 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5221 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5281 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5341 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5401 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5461 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5521 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5581 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5641 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5701 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5761 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5821 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5881 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5941 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6001 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6061 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6121 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6181 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcgac attgattatt
     6241 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     6301 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     6361 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     6421 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     6481 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     6541 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     6601 taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
     6661 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggc
//
