LOCUS       sequence-413601-        6843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses untagged ATG3 (H266A) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6843)
  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 6843)
  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 (H266A)
FEATURES             Location/Qualifiers
     source          1..6843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(35..608,609..611,612..620)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(202..219)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     429..448
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(621..623,624..626,627..1337)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(666..687)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(927..946)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1274..1295
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             1404..1997
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2063..2080)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2234..2822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2314..2333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2986..3777,3778..3846))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3609..3628
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3847..3951)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             3966..4559
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    4622..4979
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5044..5460
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5390..5412
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5428..5444
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    5470..5844
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     5782..5798
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
ORIGIN
        1 gggccggcgc gccgcggccg ctacgtaaat tccgcccctc tccctccccc ccccctaacg
       61 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
      121 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
      181 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
      241 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
      301 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
      361 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
      421 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
      481 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
      541 ggttaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
      601 atgataagct tgccacaacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
      661 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
      721 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
      781 tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc
      841 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
      901 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
      961 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1021 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     1081 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     1141 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     1201 tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg
     1261 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1321 tggacgagct gtacaagagc ggcccagcca cagtggtcga cgataaaata aaagatttta
     1381 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     1441 taagtaacgc cattttggaa ggcatggaaa aatacataac tgagaataga gaagttcaga
     1501 tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc
     1561 agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg
     1621 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg
     1681 cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac
     1741 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
     1801 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gggcgccagt
     1861 cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca
     1921 tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc
     1981 agcgggggtc tttcattctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2041 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2101 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2161 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2221 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2281 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2341 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2401 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2461 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2521 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2581 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2641 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     2701 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2761 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2821 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2881 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttgcggccgg
     2941 ccgcaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3001 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3061 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3121 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3181 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3241 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3301 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3361 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3421 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3481 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3541 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3601 aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3661 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3721 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3781 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3841 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3901 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3961 tgcataatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
     4021 aaggcatgga aaaatacata actgagaata gaaaagttca gatcaaggtc aggaacagat
     4081 ggaacagctg aatatgggcc aaagcggata tctgtggtaa gcagttcctg ccccggctca
     4141 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
     4201 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
     4261 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
     4321 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
     4381 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
     4441 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
     4501 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
     4561 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
     4621 taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
     4681 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
     4741 ctgacgagtt cggaacaccc ggccgcaacc ctgggagacg tcccagggat ctcgggggcc
     4801 gtttttgtgg cccgacctga gtccaaaaat cccgatcgtt ttggactctt tggtgcaccc
     4861 cccttagagg agggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct
     4921 ccgtctgaat ttttgctttc ggtttgggac cgaagccgcg ccgcgcgtct tgtctgctgc
     4981 agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaatatgggc
     5041 ccgggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag
     5101 cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct
     5161 gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc
     5221 atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc
     5281 ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt
     5341 gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct
     5401 cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc
     5461 cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa cttccctgac
     5521 cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca ggctctctac
     5581 ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca actggaccga
     5641 ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg acaccagact
     5701 aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac ccccaccgcc
     5761 ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc tgccgacccc
     5821 gggggtggac catcctctag actgccggat ctagctagtt aattaaggat cccagtgtgg
     5881 tggtacggaa ttcatgcaga atgtgattaa tactgtgaag ggaaaggcac tggaagtggc
     5941 tgagtacctg accccggtcc tcaaggaatc aaagtttaag gaaacaggtg taattacccc
     6001 agaagagttt gtggcagctg gagatcacct agtccaccac tgtccaacat ggcaatgggc
     6061 tacaggggaa gaattgaaag tgaaggcata cctaccaaca ggcaaacaat ttttggtaac
     6121 caaaaatgtg ccgtgctata agcggtgcaa acagatggaa tattcagatg aattggaagc
     6181 tatcattgaa gaagatgatg gtgatggcgg atgggtagat acatatcaca acacaggtat
     6241 tacaggaata acggaagccg ttaaagagat cacactggaa aataaggaca atataaggct
     6301 tcaagattgc tcagcactat gtgaagagga agaagatgaa gatgaaggag aagctgcaga
     6361 tatggaagaa tatgaagaga gtggattgtt ggaaacagat gaggctaccc tagatacaag
     6421 gaaaatagta gaagcttgta aagccaaaac tgatgctggc ggtgaagatg ctattttgca
     6481 aaccagaact tatgaccttt acatcactta tgataaatat taccagactc cacgattatg
     6541 gttgtttggc tatgatgagc aacggcagcc tttaacagtt gagcacatgt atgaagacat
     6601 cagtcaggat catgtgaaga aaacagtgac cattgaaaat caccctcatc tgccaccacc
     6661 tcccatgtgt tcagttcacc catgcagggc agctgaggtg atgaagaaaa tcattgagac
     6721 tgttgcagaa ggagggggag aacttggagt tcatatgtat cttcttattt tcttgaaatt
     6781 tgtacaagct gtcattccaa caatagaata tgactacaca agacacttca caatgtaact
     6841 cga
//
