LOCUS       sequence-174999-        6259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of the mammalian Atg8 protein GABARAPL2 without a 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6259)
  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 6259)
  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: pMX-IY GBRPL2
FEATURES             Location/Qualifiers
     source          1..6259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(44..835,836..904))
                     /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     667..686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(905..1009)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             1024..1617
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    1680..2037
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             2102..2518
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2448..2470
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2486..2502
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    2528..2902
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     2840..2856
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     misc_feature    join(3349..3922,3923..3925,3926..3934)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3516..3533)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3743..3762
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3935..3937,3938..3940,3941..4654)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3980..4001)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4241..4260)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4588..4609
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4721..5314
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5380..5397)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5551..6139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5631..5650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
       61 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
      121 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
      181 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
      241 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
      301 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
      361 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
      421 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
      481 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
      541 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
      601 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
      661 cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
      721 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
      781 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
      841 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
      901 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
      961 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcctg
     1021 cataatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa
     1081 ggcatggaaa aatacataac tgagaataga aaagttcaga tcaaggtcag gaacagatgg
     1141 aacagctgaa tatgggccaa agcggatatc tgtggtaagc agttcctgcc ccggctcagg
     1201 gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
     1261 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt
     1321 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     1381 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     1441 tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc
     1501 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
     1561 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg
     1621 gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggta
     1681 agctggccag caacttatct gtgtctgtcc gattgtctag tgtctatgac tgattttatg
     1741 cgcctgcgtc ggtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact
     1801 gacgagttcg gaacacccgg ccgcaaccct gggagacgtc ccagggatct cgggggccgt
     1861 ttttgtggcc cgacctgagt ccaaaaatcc cgatcgtttt ggactctttg gtgcaccccc
     1921 cttagaggag ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc
     1981 gtctgaattt ttgctttcgg tttgggaccg aagccgcgcc gcgcgtcttg tctgctgcag
     2041 catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa atatgggccc
     2101 gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg
     2161 gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc
     2221 agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat
     2281 cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc
     2341 ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt
     2401 acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc
     2461 tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc taggcgcccc
     2521 catatggcca tatgagatct tatatggggc acccccgccc cttgtaaact tccctgaccc
     2581 tgacatgaca agagttacta acagcccctc tctccaagct cacttacagg ctctctactt
     2641 agtccagcac gaagtctgga gacctctggc ggcagcctac caagaacaac tggaccgacc
     2701 ggtggtacct cacccttacc gagtcggcga cacagtgtgg gtccgccgac accagactaa
     2761 gaacctagaa cctcgctgga aaggacctta cacagtcctg ctgaccaccc ccaccgccct
     2821 caaagtagac ggcatcgcag cttggataca cgccgcccac gtgaaggctg ccgaccccgg
     2881 gggtggacca tcctctagac tgccggatct agctagttaa ttaaggatcc cagtgtggtg
     2941 gtacggaatt ccaccatgaa gtggatgttc aaggaggacc actcgctgga acacagatgc
     3001 gtggagtccg cgaagattcg agcgaaatat cccgacaggg ttccggtgat tgtggaaaag
     3061 gtctcaggct ctcagattgt tgacattgac aaacggaagt acttggttcc atctgatatc
     3121 actgtggctc agttcatgtg gatcatcagg aaaaggatcc agcttccttc tgaaaaggcg
     3181 atcttcctgt ttgtggataa gacagtccca cagtccagcc taactatggg acagctttac
     3241 gagaaggaaa aagatgaaga tggattctta tatgtggcct acagcggaga gaacactttt
     3301 ggcttctgac tcgagggccg gcgcgccgcg gccgctacgt aaattccgcc cctctccctc
     3361 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     3421 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     3481 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     3541 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     3601 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     3661 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     3721 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     3781 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     3841 atgtgtttag tcgaggttaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     3901 ctttgaaaaa cacgatgata agcttgccac aaccatggtg agcaagggcg aggagctgtt
     3961 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     4021 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     4081 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcct
     4141 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     4201 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     4261 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     4321 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     4381 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     4441 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     4501 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag
     4561 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     4621 gatcactctc ggcatggacg agctgtacaa gtaaagcggc ccagccacag tggtcgacga
     4681 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     4741 ggtttggcaa gctagcttaa gtaacgccat tttggaaggc atggaaaaat acataactga
     4801 gaatagagaa gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca
     4861 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg
     4921 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     4981 cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc
     5041 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     5101 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     5161 tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa
     5221 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     5281 gtgattgact acccgtcagc gggggtcttt cattctgcat taatgaatcg gccaacgcgc
     5341 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5401 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5461 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5521 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5581 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5641 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5701 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5761 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5821 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5881 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5941 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6001 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6061 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6121 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6181 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6241 gatccttttg cggccggcc
//
