LOCUS       sequence-400990-        6305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of ATG14.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6305)
  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 6305)
  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-ATG14
FEATURES             Location/Qualifiers
     source          1..6305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        118..497
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        499..774
                     /label=chicken beta-actin promoter
     intron          775..1791
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1715..1737
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1799..1818
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1866..1875
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3440..3459)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3505..3560
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3559..3578)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3921..3937)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3921..3937)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3934..3956)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3945..3961
                     /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(3969..3975,3976..3993,3994..3999))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4014..4035
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4094..4289
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4140..4275
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4295..4429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4332..4351)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4386..4405
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4497..4514)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4668..5256)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4748..4767)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5427..6218,6219..6287))
                     /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     6050..6069
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6288..6305,1..87))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
       61 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcgac
      121 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      181 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      241 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      301 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      361 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      421 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      481 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
      541 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
      601 gggcgggggg ggggggggsg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg
      661 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
      721 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
      781 gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg
      841 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
      901 tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt
      961 gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt
     1021 gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg
     1081 cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt
     1141 gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg
     1201 ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct
     1261 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg
     1321 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     1381 cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt
     1441 cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga
     1501 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
     1561 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1621 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
     1681 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     1741 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     1801 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattcctcga tcgacggtat
     1861 cgatagccac catggcttcc ccatctggaa agggagctag ggctctggag gctcctggat
     1921 gcggaccacg gcccctggcc agagacctgg tggatagcgt ggacgatgct gaggggctgt
     1981 acgtggccgt ggaaagatgc cccctgtgta ataccacacg gagaaggctg acatgcgcca
     2041 agtgtgtgca gtccggggat ttcgtgtatt ttgacggacg cgatcgggag agattcattg
     2101 acaagaaaga aaggctgtcc cgcctgaagt ctaaacagga ggaatttcag aaagaggtgc
     2161 tgaaggccat ggagggcaag tggatcactg atcagctgag gtggaaaatc atgtcctgca
     2221 agatgcgcat tgagcagctg aaacagacca tctgtaaggg aaacgaggaa atggagaaaa
     2281 atagcgaagg cctgctgaag acaaaagaaa agaatcagaa gctgtactcc cgggctcaga
     2341 gacaccagga gaagaaagaa aaaattcaga ggcataaccg caagctgggc gatctggtgg
     2401 agaagaaaac catcgacctg aggtctcact atgaacgcct ggccaatctg cgccggagcc
     2461 atatcctgga gctgacatcc gtgatcttcc caattgagga agtgaaaact ggagtgaggg
     2521 accccgctga tgtgagctcc gagagtgact cagccatgac ctctagtaca gtgagtaagc
     2581 tggctgaagc cagaaggact acctacctgt caggccgctg ggtgtgcgac gatcacaacg
     2641 gcgatacaag catctccatt actgggccct ggattagcct gcctaacaat ggcgactata
     2701 gtgcctacta ttcatgggtg gaggaaaaga aaacaactca ggggccagat atggagcagt
     2761 ctaatcccgc ttacactatc agtgccgctc tgtgctatgc cacccagctg gtgaacatcc
     2821 tgtctcatat tctggacgtg aacctgccca agaaactgtg caattcagag ttctgtggcg
     2881 aaaacctgag caaacagaag tttactaggg ctgtgaagaa actgaacgcc aatattctgt
     2941 acctgtgttt tagccagcac gtgaatctgg atcagctgca gcctctgcat accctgcgca
     3001 acctgatgta tctggtgtcc ccttcaagcg agcacctggg aaggtctgga ccattcgaag
     3061 tgagagctga tctggaggaa agcatggagt ttgtggaccc tggagtggcc ggcgagtctg
     3121 acgaaagtgg ggatgagcgg gtgtctgacg aggaaactga tctgggaacc gactgggaaa
     3181 acctgcctag tccaagattc tgtgacatcc catctcagag tgtggaggtg tcacagagcc
     3241 agtccacaca ggcttccccc cctattgctt cctctagtgc tggaggaatg atctcaagcg
     3301 ccgctgcctc agtgaccagc tggtttaagg cctacacagg ccataggtga ctcgaggaat
     3361 tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg
     3421 ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     3481 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     3541 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     3601 agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca
     3661 aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat
     3721 tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt
     3781 tttctttaac atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct
     3841 cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc
     3901 ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3961 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4021 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4081 tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4141 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4201 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4261 gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg
     4321 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4381 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa
     4441 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4501 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4561 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4621 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4681 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4741 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4801 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4861 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4921 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4981 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5041 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5101 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5161 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5221 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5281 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5341 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5401 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5461 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5521 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5581 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5641 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5701 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5761 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5821 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5881 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5941 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6001 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6061 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6121 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6181 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6241 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6301 caata
//
