LOCUS       sequence-422852-        6889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses untagged ATG3 (K62D/K64D) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6889)
  AUTHORS   Rao S, Skulsuppaisarn M, Strong LM, Ren X, Lazarou M, Hurley JH, 
            Hummer G
  TITLE     Three-step docking by WIPI2, ATG16L1, and ATG3 delivers LC3 to the 
            phagophore.
  JOURNAL   Sci Adv. 2024 Feb 9;10(6):eadj8027. doi: 10.1126/sciadv.adj8027. 
            Epub 2024 Feb 7.
  PUBMED    38324698
REFERENCE   2  (bases 1 to 6889)
  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: "2024-02-9"; volume: "10(6):eadj8027. doi"; pages: " 
            10.1126/sciadv.adj8027. Epub 2024 Feb 7"
COMMENT     Sequence Label: pMX-IG-ATG3 (K62D/K64D)
FEATURES             Location/Qualifiers
     source          1..6889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    31..597
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    47..599
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(191..208)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     418..437
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(611..613,614..616,617..1330)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(656..677)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(917..936)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1264..1285
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             1501..1970
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2100..2117)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2271..2859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2351..2370)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3023..3814,3815..3883))
                     /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     3646..3665
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3884..3988)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             4018..4611
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    4674..5031
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5090..5506
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5436..5458
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5474..5490
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    5516..5890
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     5828..5844
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
ORIGIN
        1 gggccggcgc gccgcggccg ctacgtaaat tccgcccccc cccctaacgt tactggccga
       61 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
      121 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
      181 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
      241 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
      301 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
      361 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
      421 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
      481 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
      541 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
      601 ggccacaacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      661 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      721 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      781 ctggcccacc ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga
      841 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      901 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
      961 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1021 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     1081 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     1141 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1201 cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga
     1261 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1321 gtacaagtaa gtcgacgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     1381 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     1441 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag
     1501 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     1561 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     1621 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
     1681 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     1741 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     1801 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
     1861 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     1921 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca catgcagcat
     1981 gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc atagttgcat
     2041 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2101 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2161 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2221 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2281 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2341 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2401 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2461 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2521 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2581 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2641 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2701 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2761 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2821 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2881 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2941 tcaaaaagga tcttcaccta gatccttttg cggccggccg caaatcaatc taaagtatat
     3001 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3061 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3121 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3181 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3241 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3301 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3361 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3421 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3481 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3541 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3601 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3661 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3721 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3781 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3841 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3901 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     3961 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgcataat
     4021 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     4081 gaaaaataca taactgagaa tagaaaagtt cagatcaagg tcaggaacag atggaacagc
     4141 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     4201 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     4261 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     4321 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     4381 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     4441 aaagagccca caacccctca ctcggcgcgc cagtcctccg attgactgag tcgcccgggt
     4501 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     4561 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat ttgggggctc
     4621 gtccgggatc gggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg
     4681 ccagcaactt atctgtgtct gtccgattgt ctagtgtcta tgactgattt tatgcgcctg
     4741 cgtcggtact agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag
     4801 ttcggaacac ccggccgcaa ccctgggaga cgtcccaggg acttcggggg ccgtttttgt
     4861 ggcccgacct gagtccaaaa atcccgatcg ttttggactc tttggtgcac ccccctaata
     4921 ggagggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc ctccgtctga
     4981 atttttgctt tcggtttggg accgaagccg cgccgcgcgt cttgtctgct gcagcatcgt
     5041 tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct gaaaattagg gccagactgt
     5101 taccactccc ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa
     5161 ccagtcggta gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac
     5221 ctttaacgtc ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa
     5281 gatcaaggtc ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac
     5341 ctgggaagcc ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc
     5401 tccgcctcct cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc
     5461 gcctcgatcc tccctttatc cagccctcac tccttctcta ggcgccccca tatggccata
     5521 tgagatctta tatggggcac ccccgcccct tgtaaacttc cctgaccctg acatgacaag
     5581 agttactaac agcccctctc tccaagctca cttacaggct ctctacttag tccagcacga
     5641 agtctggaga cctctggcgg cagcctacca agaacaactg gaccgaccgg tggtacctca
     5701 cccttaccga gtcggcgaca cagtgtgggt ccgccgacac cagactaaga acctagaacc
     5761 tcgctggaaa ggaccttaca cagtcctgct gaccaccccc accgccctca aagtagacgg
     5821 catcgcagct tggatacacg ccgcccacgt gaaggctgcc gaccccgggg gtggaccatc
     5881 ctctagactg ccggatctag ctagttaatt aaggatccca gtgtggtggt acggaattca
     5941 tgcagaatgt gattaatact gtgaagggaa aggcactgga agtggctgag tacctgaccc
     6001 cggtcctcaa ggaatcaaag tttaaggaaa caggtgtaat taccccagaa gagtttgtgg
     6061 cagctggaga tcacctagtc caccactgtc caacatggca atgggctaca ggggaagaat
     6121 tggacgtgga cgcataccta ccaacaggca aacaattttt ggtaaccaaa aatgtgccgt
     6181 gctataagcg gtgcaaacag atggaatatt cagatgaatt ggaagctatc attgaagaag
     6241 atgatggtga tggcggatgg gtagatacat atcacaacac aggtattaca ggaataacgg
     6301 aagccgttaa agagatcaca ctggaaaata aggacaatat aaggcttcaa gattgctcag
     6361 cactatgtga agaggaagaa gatgaagatg aaggagaagc tgcagatatg gaagaatatg
     6421 aagagagtgg attgttggaa acagatgagg ctaccctaga tacaaggaaa atagtagaag
     6481 cttgtaaagc caaaactgat gctggcggtg aagatgctat tttgcaaacc agaacttatg
     6541 acctttacat cacttatgat aaatattacc agactccacg attatggttg tttggctatg
     6601 atgagcaacg gcagccttta acagttgagc acatgtatga agacatcagt caggatcatg
     6661 tgaagaaaac agtgaccatt gaaaatcacc ctcatctgcc accacctccc atgtgttcag
     6721 ttcacccatg caggcatgct gaggtgatga agaaaatcat tgagactgtt gcagaaggag
     6781 ggggagaact tggagttcat atgtatcttc ttattttctt gaaatttgta caagctgtca
     6841 ttccaacaat agaatatgac tacacaagac acttcacaat gtaactcga
//
