LOCUS       sequence-174993-        4915 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  For expression of the mammalian Atg8 protein GABARAP tagged with HA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4915)
  AUTHORS   Nguyen TN, Padman BS, Usher J, Oorschot V, Ramm G, Lazarou M
  TITLE     Atg8 family LC3/GABARAP proteins are crucial for 
            autophagosome-lysosome fusion but not autophagosome formation during
            PINK1/Parkin mitophagy and starvation.
  JOURNAL   J Cell Biol. 2016 Dec 19;215(6):857-874. Epub 2016 Nov 18.
  PUBMED    27864321
REFERENCE   2  (bases 1 to 4915)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol."; date: "2016-12-19"; volume: "215(6)"; pages: "857-874. Epub 
            2016 Nov 18"
COMMENT     Sequence Label: pBMN HA-GBRP
FEATURES             Location/Qualifiers
     source          1..4915
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     LTR             1..470
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    535..892
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             957..1373
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1303..1325
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1341..1357
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1383..1757
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1695..1711
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             1764..1790
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             2315..2784
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2914..2931)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3085..3673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3165..3184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3837..4628,4629..4697))
                     /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     4460..4479
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4698..4802)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
       61 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      121 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      181 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      241 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      301 aaaagagccc acaacccctc actcggcgcg ccagtcctcc gattgactga gtcgcccggg
      361 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      421 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      481 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg
      541 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      601 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      661 gttcggaaca cccggccgca accctgggag acgtcccagg gacttcgggg gccgtttttg
      721 tggcccgacc tgagtccaaa aatcccgatc gttttggact ctttggtgca ccccccttag
      781 aggagggata tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg
      841 aatttttgct ttcggtttgg gaccgaagcc gcgccgcgcg tcttgtctgc tgcagcatcg
      901 ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaatatg ggcccgggcc
      961 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
     1021 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
     1081 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     1141 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     1201 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
     1261 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
     1321 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gcccccatat
     1381 ggccatatga gatcttatat ggggcacccc cgccccttgt aaacttccct gaccctgaca
     1441 tgacaagagt tactaacagc ccctctctcc aagctcactt acaggctctc tacttagtcc
     1501 agcacgaagt ctggagacct ctggcggcag cctaccaaga acaactggac cgaccggtgg
     1561 tacctcaccc ttaccgagtc ggcgacacag tgtgggtccg ccgacaccag actaagaacc
     1621 tagaacctcg ctggaaagga ccttacacag tcctgctgac cacccccacc gccctcaaag
     1681 tagacggcat cgcagcttgg atacacgccg cccacgtgaa ggctgccgac cccgggggtg
     1741 gaccatcctc tagactgccg atgtacccat acgatgttcc agattacgct aagttcgtgt
     1801 acaaagaaga gcatccgttc gagaagcgcc gctctgaggg cgagaagatc cgaaagaaat
     1861 acccggaccg ggtgccggtg atagtagaaa aggctcccaa agctcggata ggagacctgg
     1921 acaaaaagaa atacctggtg ccttctgatc tcacagttgg tcagttctac ttcttgatcc
     1981 ggaagcgaat tcatctccga gctgaggatg ccttgttttt ctttgtcaac aatgtcattc
     2041 cacccaccag tgccacaatg ggtcagctgt accaggaaca ccatgaagaa gacttctttc
     2101 tctacattgc ctacagtgac gaaagtgtct acggtctgtg ataagtcgac gataaaataa
     2161 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     2221 aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag
     2281 aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct
     2341 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg
     2401 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     2461 ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     2521 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     2581 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     2641 ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt
     2701 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     2761 ctacccgtca gcgggggtct ttcacatgca gcatgtatca aaattaattt ggtttttttt
     2821 cttaagtatt tacattaaat ggccatagtt gcattaatga atcggccaac gcgcggggag
     2881 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2941 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3001 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3061 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3121 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3181 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3241 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3301 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3361 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3421 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3481 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3541 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3601 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3661 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3721 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3781 tttgcggccg gccgcaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3841 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3901 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3961 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4021 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4081 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4141 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4201 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4261 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4321 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4381 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4441 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4501 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4561 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4621 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4681 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4741 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4801 cgcacatttc cccgaaaagt gccaccagct ttgctcttag gagtttccta atacatccca
     4861 aactcaaata tataaagcat ttgacttgtt ctatgcccta gggggcgggg ggaag
//
