LOCUS       sequence-401051-        7045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mCherry tagged ATG14.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7045)
  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 7045)
  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-mCherry-ATG14
FEATURES             Location/Qualifiers
     source          1..7045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        123..502
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        504..779
                     /label=chicken beta-actin promoter
     intron          780..1796
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1720..1742
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1804..1823
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1870..1879
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1876..2583
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2021..2039)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2289..2308
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2593..2613
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(4185..4204)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4250..4305
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4304..4323)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4666..4682)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4666..4682)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4679..4701)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4690..4706
                     /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(4714..4720,4721..4738,4739..4744))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4759..4780
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4839..5034
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4885..5020
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5040..5174
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5077..5096)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5131..5150
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5242..5259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5413..6001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5493..5512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6172..6963,6964..7032))
                     /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     6795..6814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7033..7045,1..92))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
       61 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctggg
      121 tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
      181 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      241 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      301 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      361 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      421 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      481 attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat
      541 ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc
      601 gatgggggcg gggggggggg gggsgcgcgc caggcggggc ggggcggggc gaggggcggg
      661 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      721 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      781 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      841 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      901 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      961 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
     1021 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
     1081 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
     1141 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1201 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
     1261 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
     1321 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
     1381 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
     1441 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1501 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     1561 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1621 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1681 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1741 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1801 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc ctcgatcgac
     1861 ggtatcgatg ccaccatggt gagcaagggc gaggaggata acatggccat catcaaggag
     1921 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
     1981 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     2041 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     2101 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     2161 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     2221 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     2281 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg
     2341 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     2401 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     2461 ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc
     2521 gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     2581 aagccaacga ccgaaaacct gtattttcag ggtggtatgg cttccccatc tggaaaggga
     2641 gctagggctc tggaggctcc tggatgcgga ccacggcccc tggccagaga cctggtggat
     2701 agcgtggacg atgctgaggg gctgtacgtg gccgtggaaa gatgccccct gtgtaatacc
     2761 acacggagaa ggctgacatg cgccaagtgt gtgcagtccg gggatttcgt gtattttgac
     2821 ggacgcgatc gggagagatt cattgacaag aaagaaaggc tgtcccgcct gaagtctaaa
     2881 caggaggaat ttcagaaaga ggtgctgaag gccatggagg gcaagtggat cactgatcag
     2941 ctgaggtgga aaatcatgtc ctgcaagatg cgcattgagc agctgaaaca gaccatctgt
     3001 aagggaaacg aggaaatgga gaaaaatagc gaaggcctgc tgaagacaaa agaaaagaat
     3061 cagaagctgt actcccgggc tcagagacac caggagaaga aagaaaaaat tcagaggcat
     3121 aaccgcaagc tgggcgatct ggtggagaag aaaaccatcg acctgaggtc tcactatgaa
     3181 cgcctggcca atctgcgccg gagccatatc ctggagctga catccgtgat cttcccaatt
     3241 gaggaagtga aaactggagt gagggacccc gctgatgtga gctccgagag tgactcagcc
     3301 atgacctcta gtacagtgag taagctggct gaagccagaa ggactaccta cctgtcaggc
     3361 cgctgggtgt gcgacgatca caacggcgat acaagcatct ccattactgg gccctggatt
     3421 agcctgccta acaatggcga ctatagtgcc tactattcat gggtggagga aaagaaaaca
     3481 actcaggggc cagatatgga gcagtctaat cccgcttaca ctatcagtgc cgctctgtgc
     3541 tatgccaccc agctggtgaa catcctgtct catattctgg acgtgaacct gcccaagaaa
     3601 ctgtgcaatt cagagttctg tggcgaaaac ctgagcaaac agaagtttac tagggctgtg
     3661 aagaaactga acgccaatat tctgtacctg tgttttagcc agcacgtgaa tctggatcag
     3721 ctgcagcctc tgcataccct gcgcaacctg atgtatctgg tgtccccttc aagcgagcac
     3781 ctgggaaggt ctggaccatt cgaagtgaga gctgatctgg aggaaagcat ggagtttgtg
     3841 gaccctggag tggccggcga gtctgacgaa agtggggatg agcgggtgtc tgacgaggaa
     3901 actgatctgg gaaccgactg ggaaaacctg cctagtccaa gattctgtga catcccatct
     3961 cagagtgtgg aggtgtcaca gagccagtcc acacaggctt ccccccctat tgcttcctct
     4021 agtgctggag gaatgatctc aagcgccgct gcctcagtga ccagctggtt taaggcctac
     4081 acaggccata ggtgactcga ggaattcact cctcaggtgc aggctgccta tcagaaggtg
     4141 gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc
     4201 aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa
     4261 tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat
     4321 gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat
     4381 gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa
     4441 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
     4501 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
     4561 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
     4621 ttatggagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc
     4681 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4741 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4801 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc
     4861 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4921 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4981 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta
     5041 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5101 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5161 atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5221 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5281 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5341 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5401 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5461 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5521 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5581 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5641 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5701 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5761 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5821 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5881 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5941 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6001 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6061 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6121 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6181 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6241 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6301 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6361 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6421 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6481 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6541 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6601 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6661 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6721 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6781 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6841 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6901 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6961 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7021 ttgaatactc atactcttcc ttttt
//
