LOCUS       sequence-401054-        5744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of GST tagged ATG13 (450-517).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5744)
  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 5744)
  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-GST-ATG13 (450-517)
FEATURES             Location/Qualifiers
     source          1..5744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        111..490
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        492..767
                     /label=chicken beta-actin promoter
     intron          768..1784
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1708..1730
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1792..1811
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1859..1868
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1865..2521
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MASPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
                     KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIR
                     YGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVV
                     LYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2479..2501
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2549..2569
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2872..2891)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2937..2992
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2991..3010)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3353..3369)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3353..3369)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3366..3388)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3377..3393
                     /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(3401..3407,3408..3425,3426..3431))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3446..3467
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3526..3721
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3572..3707
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3727..3861
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3764..3783)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3818..3837
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3929..3946)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4100..4688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4180..4199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4859..5650,5651..5719))
                     /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     5482..5501
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5720..5744,1..80))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
       61 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgggtc gacattgatt
      121 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      181 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      241 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      301 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      361 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      421 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      481 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
      541 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
      601 gggggggggg gsgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc
      661 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
      721 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
      781 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
      841 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
      901 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc
      961 tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg
     1021 tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg
     1081 cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc
     1141 ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt
     1201 gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag
     1261 ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg
     1321 ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg
     1381 ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg
     1441 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt
     1501 tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc
     1561 gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt
     1621 cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct
     1681 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     1741 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     1801 tggttattgt gctgtctcat cattttggca aagaattcct cgatcgacgg tatcgatagc
     1861 caccatggca tcccctatac taggttattg gaaaattaag ggccttgtgc aacccactcg
     1921 acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg
     1981 tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc ttccttatta
     2041 tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata tagctgacaa
     2101 gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc
     2161 ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact ttgaaactct
     2221 caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag atcgtttatg
     2281 tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt tgtatgacgc
     2341 tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa aattagtttg
     2401 ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat ccagcaagta
     2461 tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc atcctccaaa
     2521 atcggatgat tacgatatcc caacgaccga aaacctgtat tttcagggtg gtaccaaacc
     2581 cgctttttcc aaggacgata tcctccctat ggatctggga accttctaca gggagtttca
     2641 gaatccccct cagctcagta gcctgagcat tgacatcggt gcccagtcca tggctgagga
     2701 cctcgattct ctgcctgaaa aactggctgt gcacgagaag aacgtcagag aattcgacgc
     2761 atttgtggag accctgcagt gactcgagga attcactcct caggtgcagg ctgcctatca
     2821 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc
     2881 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
     2941 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     3001 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc
     3061 aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta
     3121 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
     3181 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt
     3241 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc
     3301 tcttctctta tggagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc
     3361 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3421 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3481 cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc
     3541 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3601 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3661 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3721 aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3781 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3841 gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3901 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3961 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4021 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4081 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4141 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4201 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4261 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4321 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4381 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4441 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4501 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4561 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4621 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4681 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4741 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4801 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4861 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4921 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4981 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5041 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5101 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5161 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5221 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5281 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5341 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5401 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5461 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5521 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5581 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5641 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5701 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttga
//
