LOCUS       sequence-337513-        7126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the purifcation of human ULK1 complex .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7126)
  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 7126)
  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: GFP-ATG13
FEATURES             Location/Qualifiers
     source          1..7126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..663
                     /label=chicken beta-actin promoter
     intron          664..1680
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1604..1626
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1688..1707
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1754..1763
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1760..1762,1763..1765,1766..2476)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1805..1826)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2066..2085)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2413..2434
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3006..3017
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(4148..4167)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4213..4268
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4267..4286)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4629..4645)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4629..4645)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4642..4664)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4653..4669
                     /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(4677..4683,4684..4701,4702..4707))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4722..4743
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4802..4997
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4848..4983
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5003..5137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5040..5059)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5094..5113
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5205..5222)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5376..5964)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5456..5475)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6135..6926,6927..6995))
                     /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     6758..6777
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6996..7100)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gcgccrggsg gggsggggsg gggsgrgggg
      541 sggggsgggg sgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
      601 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
      661 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
      721 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      781 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
      841 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
      961 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     1021 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     1081 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
     1141 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
     1201 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg
     1261 gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc
     1321 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     1381 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     1441 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1501 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     1561 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     1621 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     1681 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcctcgat
     1741 cgacggtatc gatgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1801 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1861 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1921 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     1981 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2041 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2101 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2161 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     2221 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     2281 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2341 gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga
     2401 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2461 ggacgagctg tacaagggta ccgagaccga cctcaacagc caggaccgta aagacctcga
     2521 caagttcatt aagtttttcg ccctcaagac cgtgcaggtg atcgtgcagg cccggctggg
     2581 agagaaaatc tgcacacgca gctcctcttc acccaccggt tctgactggt tcaacctggc
     2641 tattaaggat atccctgaag tgactcacga agcaaagaaa gcactcgctg gacagctgcc
     2701 agcagtgggt cgttcaatgt gtgtcgagat ctccctgaag acatctgaag gagactcaat
     2761 ggagctcgaa atttggtgcc tggagatgaa cgaaaagtgt gataaagaga tcaaagtgag
     2821 ctacaccgtc tataatcgtc tgtccctgct cctgaaatct ctcctggcca tcactcgggt
     2881 gacacctgct taccgcctca gtaggaagca gggtcatgaa tatgtgattc tgtacaggat
     2941 ctatttcggc gaggtccagc tctccggact gggtgaaggc tttcagactg tgcgtgtcgg
     3001 aaccgtggga actccagtcg gcacaattac cctctcttgc gcataccgta tcaatctggc
     3061 cttcatgtca accagacagt ttgagcgtac tccccctatt atgggaatca ttatcgacca
     3121 cttcgtggat agaccatacc ccagtagctc ccctatgcat ccatgcaact atcgtacagc
     3181 cggtgaagac accggcgtga tctaccccag tgtcgaggat agccaggaag tgtgtaccac
     3241 ttccttctct acctcaccac cctcacagct ctcttcaagt cgcctgagtt atcagcctgc
     3301 cgctctcgga gtcggtagcg cagacctggc ctacccagtg gtcttcgcag caggcctcaa
     3361 cgcaacccac ccacatcagc tgatggtgcc aggaaaggag ggtggagtcc cactggctcc
     3421 taatcagcca gtgcacggta ctcaggctga ccaggaaagg ctggcaactt gcacaccatc
     3481 cgatagaact cattgtgctg caacacccag ctcctctgag gacaccgaaa ctgtgagcaa
     3541 ctcaagtgag ggtcgcgcct ccccacacga tgtcctggaa accatcttcg tgagaaaagt
     3601 cggcgctttt gtgaacaagc ccattaatca ggtcacactc acctctctgg acatcccttt
     3661 cgctatgttt gcaccaaaga acctcgagct ggaagacact gatcccatgg tgaatcctcc
     3721 agactcaccc gagacagaaa gtcctctcca gggcagtctg cacagcgatg gaagctccgg
     3781 tggctcttca ggcaacacac atgacgattt cgtcatgatt gacttcaaac ccgctttttc
     3841 caaggacgat atcctcccta tggatctggg aaccttctac agggagtttc agaatccccc
     3901 tcagctcagt agcctgagca ttgacatcgg tgcccagtcc atggctgagg acctcgattc
     3961 tctgcctgaa aaactggctg tgcacgagaa gaacgtcaga gaattcgacg catttgtgga
     4021 gaccctgcag taacccgggt gatatcttaa ttaattaact cgaggaattc actcctcagg
     4081 tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc
     4141 actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     4201 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     4261 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     4321 tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat
     4381 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     4441 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
     4501 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
     4561 cagtcatagc tgtccctctt ctcttatgga gatccctcga cctgcagccc aagcttggcg
     4621 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4681 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4741 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc
     4801 cgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4861 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4921 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4981 gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag
     5041 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5101 gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa
     5161 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5221 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5281 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5341 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5401 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5461 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5521 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5581 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5641 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5701 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5761 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5821 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5881 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5941 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6001 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6061 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6121 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6181 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6241 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6301 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6361 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6421 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6481 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6541 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6601 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6661 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6721 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6781 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6841 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6901 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6961 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7021 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7081 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctg
//
