LOCUS       sequence-408594-        5834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of GST tagged ATG13 (363-460).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5834)
  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 5834)
  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 (363-460)
FEATURES             Location/Qualifiers
     source          1..5834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        97..476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        478..753
                     /label=chicken beta-actin promoter
     intron          754..1770
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1694..1716
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1778..1797
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1845..1854
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1851..2507
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MASPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
                     KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIR
                     YGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVV
                     LYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2465..2487
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2535..2555
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2948..2967)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3013..3068
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3067..3086)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3429..3445)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3429..3445)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3442..3464)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3453..3469
                     /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(3477..3483,3484..3501,3502..3507))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3522..3543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3602..3797
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3648..3783
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3803..3937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3840..3859)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3894..3913
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4005..4022)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4176..4764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4256..4275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4935..5726,5727..5795))
                     /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     5558..5577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5796..5834,1..66))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
       61 tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca ttgattattg actagttatt
      121 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      181 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      241 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      301 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      361 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      421 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
      481 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
      541 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc
      601 gcgccrgggg gggggggggg gggsgrgggg sggggcgggg cgaggcggag aggtgcggcg
      661 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
      721 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc
      781 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
      841 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa
      901 tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct
      961 ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg
     1021 tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg
     1081 ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc
     1141 tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg
     1201 ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc
     1261 ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg
     1321 gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg
     1381 ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc
     1441 cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt
     1501 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg
     1561 tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt
     1621 ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga
     1681 cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac
     1741 catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg
     1801 tctcatcatt ttggcaaaga attcctcgat cgacggtatc gatagccacc atggcatccc
     1861 ctatactagg ttattggaaa attaagggcc ttgtgcaacc cactcgactt cttttggaat
     1921 atcttgaaga aaaatatgaa gagcatttgt atgagcgcga tgaaggtgat aaatggcgaa
     1981 acaaaaagtt tgaattgggt ttggagtttc ccaatcttcc ttattatatt gatggtgatg
     2041 ttaaattaac acagtctatg gccatcatac gttatatagc tgacaagcac aacatgttgg
     2101 gtggttgtcc aaaagagcgt gcagagattt caatgcttga aggagcggtt ttggatatta
     2161 gatacggtgt ttcgagaatt gcatatagta aagactttga aactctcaaa gttgattttc
     2221 ttagcaagct acctgaaatg ctgaaaatgt tcgaagatcg tttatgtcat aaaacatatt
     2281 taaatggtga tcatgtaacc catcctgact tcatgttgta tgacgctctt gatgttgttt
     2341 tatacatgga cccaatgtgc ctggatgcgt tcccaaaatt agtttgtttt aaaaaacgta
     2401 ttgaagctat cccacaaatt gataagtact tgaaatccag caagtatata gcatggcctt
     2461 tgcagggctg gcaagccacg tttggtggtg gcgaccatcc tccaaaatcg gatgattacg
     2521 atatcccaac gaccgaaaac ctgtattttc agggtggtac ccacgatgtc ctggaaacca
     2581 tcttcgtgag aaaagtcggc gcttttgtga acaagcccat taatcaggtc acactcacct
     2641 ctctggacat ccctttcgct atgtttgcac caaagaacct cgagctggaa gacactgatc
     2701 ccatggtgaa tcctccagac tcacccgaga cagaaagtcc tctccagggc agtctgcaca
     2761 gcgatggaag ctccggtggc tcttcaggca acacacatga cgatttcgtc atgattgact
     2821 tcaaacccgc tttttccaag gacgatatcc tcccttgact cgaggaattc actcctcagg
     2881 tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc
     2941 actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     3001 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     3061 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     3121 tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat
     3181 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     3241 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
     3301 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
     3361 cagtcatagc tgtccctctt ctcttatgga gatccctcga cctgcagccc aagcttggcg
     3421 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3481 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3541 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc
     3601 cgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3661 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3721 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3781 gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag
     3841 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3901 gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa
     3961 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4021 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4081 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4141 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4201 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4261 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4321 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4381 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4441 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4501 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4561 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4621 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4681 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4741 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4801 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4861 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4921 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4981 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5041 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5101 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5161 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5221 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5281 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5341 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5401 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5461 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5521 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5581 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5641 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5701 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5761 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5821 agcatttatc aggg
//
