LOCUS       sequence-212742-        7352 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of hATG5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7352)
  AUTHORS   Kim M, Sandford E, Gatica D, Qiu Y, Liu X, Zheng Y, Schulman BA, Xu 
            J, Semple I, Ro SH, Kim B, Mavioglu RN, Tolun A, Jipa A, Takats S, 
            Karpati M, Li JZ, Yapici Z, Juhasz G, Lee JH, Klionsky DJ, 
            Burmeister M
  TITLE     Mutation in ATG5 reduces autophagy and leads to ataxia with 
            developmental delay
  JOURNAL   Elife. 2016 Jan 26;5. pii: e12245. doi: 10.7554/eLife.12245.
  PUBMED    26812546
REFERENCE   2  (bases 1 to 7352)
  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: "Elife."; 
            date: "2016-01-26"; volume: "5. pii: e12245. doi"; pages: " 
            10.7554/eLife.12245"
COMMENT     Sequence Label: pLU-CMV-Flag-hATG5 WT
FEATURES             Location/Qualifiers
     source          1..7352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    92..208
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        231..534
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        535..738
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     689..709
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     735..759
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      813..822
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             822..845
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1704..2292
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1757..1777)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2175..2186)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2379..2612
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2684..2805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2721..2740)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2775..2794
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2845..2980
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3012..3028)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3012..3028)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3025..3047)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3036..3052
                     /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(3060..3066,3067..3084,3085..3090))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3105..3126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3243..3260)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3414..4002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3494..3513)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4173..4964,4965..5033))
                     /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     4796..4815
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5034..5138)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5206..5224
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5262..5284)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5384..5403
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5597..5619
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5611..5628
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5612..5628
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5643..5869
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5870..6050
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6097..6222
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6715..6948
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7133..7177
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat
       61 tagtgaacgg atctcgacgg tatcggttaa cttttaaaag aaaagggggg attggggggt
      121 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
      181 aaaaacaaat tacaaaattc aaaattttat cgataagctt gggagttccg cgttacataa
      241 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      301 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      361 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      421 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      481 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      541 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      601 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      661 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      721 gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
      781 tgttttgacc tccatagaag acaccgactc tagccaccat ggactacaaa gacgatgacg
      841 acaagtctag aatgacagat gacaaagatg tgcttcgaga tgtgtggttt ggacgaattc
      901 caacttgttt cacgctatat caggatgaga taactgaaag ggaagcagaa ccatactatt
      961 tgcttttgcc aagagtaagt tatttgacgt tggtaactga caaagtgaaa aagcactttc
     1021 agaaggttat gagacaagaa gacattagtg agatatggtt tgaatatgaa ggcacaccac
     1081 tgaaatggca ttatccaatt ggtttgctat ttgatcttct tgcatcaagt tcagctcttc
     1141 cttggaacat cacagtacat tttaagagtt ttccagaaaa agaccttctg cactgtccat
     1201 ctaaggatgc aattgaagct cattttatgt catgtatgaa agaagctgat gctttaaaac
     1261 ataaaagtca agtaatcaat gaaatgcaga aaaaagatca caagcaactc tggatgggat
     1321 tgcaaaatga cagatttgac cagttttggg ccatcaatcg gaaactcatg gaatatcctg
     1381 cagaagaaaa tggatttcgt tatatcccct ttagaatata tcagacaacg actgaaagac
     1441 ctttcattca gaagctgttt cgtcctgtgg ctgcagatgg acagttgcac acactaggag
     1501 atctcctcaa agaagtttgt ccttctgcta ttgatcctga agatggggaa aaaaagaatc
     1561 aagtgatgat tcatggaatt gagccaatgt tggaaacacc tctgcagtgg ctgagtgaac
     1621 atctgagcta cccggataat tttcttcata ttagtatcat cccacagcca acagattgac
     1681 tcgaggaatt cggatccgtc gacaatcaac ctctggatta caaaatttgt gaaagattga
     1741 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     1801 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     1861 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     1921 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     1981 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     2041 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     2101 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     2161 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     2221 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     2281 ccgcctcccc gcctggaatt cgagctcggt acctttaaga ccaatgactt acaaggcagc
     2341 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     2401 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     2461 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     2521 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     2581 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     2641 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     2701 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2761 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     2821 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2881 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     2941 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg cgggcccaaa
     3001 ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3061 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3121 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3181 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3241 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3301 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3361 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3421 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3481 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3541 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3601 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3661 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3721 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3781 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3841 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3901 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3961 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4021 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4081 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4141 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4201 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4261 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4321 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4381 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4441 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4501 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4561 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4621 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4681 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4741 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4801 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4861 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4921 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4981 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5041 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5101 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5161 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     5221 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     5281 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     5341 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     5401 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     5461 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     5521 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     5581 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctgacg
     5641 cgtgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
     5701 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
     5761 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
     5821 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
     5881 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     5941 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     6001 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
     6061 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
     6121 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
     6181 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
     6241 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
     6301 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
     6361 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
     6421 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
     6481 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
     6541 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
     6601 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     6661 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     6721 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     6781 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     6841 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     6901 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     6961 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     7021 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     7081 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     7141 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     7201 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     7261 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     7321 tcagacccac ctcccaaccc cgaggggacc cg
//
