LOCUS       sequence-400963-        6779 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Make FIP200 NTD:ATG13:ULK1 complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6779)
  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 6779)
  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-FIP200 (1-640)
FEATURES             Location/Qualifiers
     source          1..6779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        112..491
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        493..768
                     /label=chicken beta-actin promoter
     intron          769..1785
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1709..1731
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1793..1812
                     /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"
     primer_bind     complement(3908..3927)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3973..4028
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4027..4046)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4389..4405)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4389..4405)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4402..4424)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4413..4429
                     /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(4437..4443,4444..4461,4462..4467))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4482..4503
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4562..4757
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4608..4743
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4763..4897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4800..4819)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4854..4873
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4965..4982)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5136..5724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5216..5235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5895..6686,6687..6755))
                     /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     6518..6537
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6756..6779,1..81))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
       61 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt cgacattgat
      121 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      181 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      241 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      301 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      361 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      421 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      481 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
      541 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg
      601 gggggggggg ggggcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
      661 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
      721 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg
      781 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
      841 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
      901 tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg
      961 ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc
     1021 gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc
     1081 gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg
     1141 cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg
     1201 tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga
     1261 gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc
     1321 gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg
     1381 gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc
     1441 gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct
     1501 ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg
     1561 cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg
     1621 tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc
     1681 tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct
     1741 agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg
     1801 ctggttattg tgctgtctca tcattttggc aaagaattcc tcgatcgacg gtatcgatgc
     1861 caccatgggt accatgaagc tctatgtgtt tctcgtgaat accggcacta ccctgacctt
     1921 tgataccgaa ctgaccgtcc agaccgtggc tgacctgaag catgctatcc agtctaaata
     1981 caagatcgca attcagcacc aggtgctggt ggtcaacggc ggagagtgca tggccgctga
     2041 tcgcagggtc tgtacctatt cagccggcac cgacactaac ccaatcttcc tctttaataa
     2101 ggagatgatt ctgtgcgata gaccccctgc tatccccaag accactttta gcactgagaa
     2161 cgacatggaa atcaaagtgg aggaatccct gatgatgcca gccgtgttcc acaccgtcgc
     2221 ttctcgcact cagctggcac tcgagatgta cgaagtggcc aagaaactgt gctctttctg
     2281 tgagggtctc gtccacgatg aacatctgca gcaccagggc tgggcagcca ttatggctaa
     2341 cctggaggac tgttcaaata gttatcagaa actgctcttc aagtttgagt ctatctactc
     2401 aaactatctg cagagcatcg aagacattaa actgaagctc acacacctgg gaaccgcagt
     2461 gagcgtcatg gccaagatcc ctctgctcga gtgcctgact aggcattcct acagagaatg
     2521 tctgggtagg ctcgatagtc tgccagaaca cgaggacagc gagaaagccg aaatgaagcg
     2581 ctctacagag ctggtgctct cacctgacat gccaaggaca accaacgaaa gtctgctcac
     2641 cagcttccca aagtccgtgg agcatgtctc ccccgacact gccgatgctg aatctggaaa
     2701 ggagattaga gaaagttgcc agagcaccgt gcaccagcag gacgagacta caatcgatac
     2761 taaggacggc gatctgccct tctttaacgt cagtctgctc gattggatca acgtccagga
     2821 cagacctaat gatgtggagt ccctggtccg gaagtgcttt gatagcatgt cccgtctgga
     2881 cccacgtatc attcgtcctt tcatcgcaga gtgtcgtcag accattgcca agctggacaa
     2941 ccagaatatg aaagctatta agggtctgga agatcggctc tacgccctgg accagatgat
     3001 cgctagctgt ggacgcctgg tgaacgagca gaaggaactc gctcagggtt tcctggcaaa
     3061 ccagaaaaga gccgagaatc tcaaggatgc atcagtgctg cctgacctgt gcctcagtca
     3121 tgccaatcag ctgatgatta tgctccagaa ccaccgcaag ctgctcgaca tcaaacagaa
     3181 gtgtaccact gctaaacagg agctggcaaa caatctccat gtgaggctga agtggtgctg
     3241 tttcgtcatg ctgcacgcag accaggatgg cgagaagctc caggccctgc tcaggctggt
     3301 gattgagctg ctcgaaagag tgaagatcgt cgaggctctg tccaccgtcc cacagatgta
     3361 ctgcctggca gtggtcgagg tggtcagacg taaaatgttt atcaagcact acagagaatg
     3421 ggcaggtgcc ctcgtgaaag atggcaagcg tctgtatgaa gccgagaaat ctaagcgtga
     3481 gtcattcggc aaactgtttc ggaagagctt cctccgtaac cggctgttcc gcggactcga
     3541 cagttggcca cccagctttt gcacccagaa acctaggaag ttcgactgtg agctgccaga
     3601 tatctccctc aaggacctgc agtttctcca gtccttctgc ccatctgagg tgcagccctt
     3661 tctgagggtc cctctgctct gtgacttcga acctctgcac cagcatgtgc tggccctcca
     3721 caacctcgtc aaggctgcac agagtctgga tgagatgagc cagacaatca ccgacctgct
     3781 cagcgaacag aagcccgggt gatatcttaa ttaattaact cgaggaattc actcctcagg
     3841 tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc
     3901 actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     3961 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     4021 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     4081 tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat
     4141 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     4201 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
     4261 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
     4321 cagtcatagc tgtccctctt ctcttatgga gatccctcga cctgcagccc aagcttggcg
     4381 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4441 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4501 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc
     4561 cgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4621 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4681 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4741 gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag
     4801 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4861 gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa
     4921 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4981 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5041 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5101 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5161 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5221 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5281 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5341 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5401 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5461 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5521 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5581 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5641 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5701 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5761 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5821 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5881 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5941 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6001 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6061 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6121 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6181 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6241 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6301 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6361 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6421 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6481 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6541 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6601 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6661 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6721 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattg
//
