LOCUS       sequence-174902-        5316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For  expression of the mammalian Atg8 protein GABARAP without a tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5316)
  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 5316)
  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: "J Cell 
            Biol."; date: "2016-12-19"; volume: "215(6)"; pages: "857-874. Epub 
            2016 Nov 18"
COMMENT     Sequence Label: pBMN GBRP
FEATURES             Location/Qualifiers
     source          1..5316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     LTR             2293..2762
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2892..2909)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3063..3651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3143..3162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3815..4606,4607..4675))
                     /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     4438..4457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4676..4780)
                     /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 gatccaccat gaagttcgtg tacaaagaag agcatccgtt
     1801 cgagaagcgc cgctctgagg gcgagaagat ccgaaagaaa tacccggacc gggtgccggt
     1861 gatagtagaa aaggctccca aagctcggat aggagacctg gacaaaaaga aatacctggt
     1921 gccttctgat ctcacagttg gtcagttcta cttcttgatc cggaagcgaa ttcatctccg
     1981 agctgaggat gccttgtttt tctttgtcaa caatgtcatt ccacccacca gtgccacaat
     2041 gggtcagctg taccaggaac accatgaaga agacttcttt ctctacattg cctacagtga
     2101 cgaaagtgtc tacggtctgt gagtcgacga taaaataaaa gattttattt agtctccaga
     2161 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
     2221 tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa
     2281 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     2341 gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     2401 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc
     2461 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     2521 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     2581 cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact
     2641 gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
     2701 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
     2761 cacatgcagc atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg
     2821 ccatagttgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2881 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2941 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3001 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3061 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3121 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3181 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3241 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3301 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3361 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3421 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3481 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3541 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3601 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3661 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3721 gtcatgagat tatcaaaaag gatcttcacc tagatccttt tgcggccggc cgcaaatcaa
     3781 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3841 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3901 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3961 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4021 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4081 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4141 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4201 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4261 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4321 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4381 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4441 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4501 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4561 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4621 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4681 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4741 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4801 caccagcttt gctcttagga gtttcctaat acatcccaaa ctcaaatata taaagcattt
     4861 gacttgttct atgccctagg gggcgggggg aagctaagcc agcttttttt aacatttaaa
     4921 atgttaattc cattttaaat gcacagatgt ttttatttca taagggtttc aatgtgcatg
     4981 aatgctgcaa tattcctgtt accaaagcta gtataaataa aaatagataa acgtggaaat
     5041 tacttagagt ttctgtcatt aacgtttcct tcctcagttg acaacataaa tgcgctgctg
     5101 agaagccagt ttgcatctgt caggatcaat ttcccattat gccagtcata ttaattacta
     5161 gtctattagt tgatttttat ttttgacata tacatgtgaa agaccccacc tgtaggtttg
     5221 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag
     5281 aaaagttcag atcaaggtca ggaacagatg gaacag
//
