LOCUS       sequence-175004-        6287 bp    DNA     circular SYN 14-APR-2025
DEFINITION  For expression of the mammalian Atg8 protein GABARAPL1 tagged with 
            HA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6287)
  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 6287)
  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: pMX-IY HA-GBRPL1
FEATURES             Location/Qualifiers
     source          1..6287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             7..600
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    663..1020
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1085..1501
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1431..1453
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1469..1485
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1511..1885
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1823..1839
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             1942..1968
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    join(2360..2933,2934..2936,2937..2945)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2527..2544)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2754..2773
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2946..2948,2949..2951,2952..3665)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2991..3012)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3252..3271)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3599..3620
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3732..4325
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4391..4408)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4562..5150)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4642..4661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5314..6105,6106..6174))
                     /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     5937..5956
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6175..6279)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcataatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg
       61 caaggcatgg aaaaatacat aactgagaat agaaaagttc agatcaaggt caggaacaga
      121 tggaacagct gaatatgggc caaagcggat atctgtggta agcagttcct gccccggctc
      181 agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      241 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca
      301 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
      361 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
      421 agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt
      481 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
      541 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt
      601 tgggggctcg tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag
      661 gtaagctggc cagcaactta tctgtgtctg tccgattgtc tagtgtctat gactgatttt
      721 atgcgcctgc gtcggtacta gttagctaac tagctctgta tctggcggac ccgtggtgga
      781 actgacgagt tcggaacacc cggccgcaac cctgggagac gtcccaggga tctcgggggc
      841 cgtttttgtg gcccgacctg agtccaaaaa tcccgatcgt tttggactct ttggtgcacc
      901 ccccttagag gagggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc
      961 tccgtctgaa tttttgcttt cggtttggga ccgaagccgc gccgcgcgtc ttgtctgctg
     1021 cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaatatggg
     1081 cccgggccag actgttacca ctcccttaag tttgacctta ggtcactgga aagatgtcga
     1141 gcggatcgct cacaaccagt cggtagatgt caagaagaga cgttgggtta ccttctgctc
     1201 tgcagaatgg ccaaccttta acgtcggatg gccgcgagac ggcaccttta accgagacct
     1261 catcacccag gttaagatca aggtcttttc acctggcccg catggacacc cagaccaggt
     1321 cccctacatc gtgacctggg aagccttggc ttttgacccc cctccctggg tcaagccctt
     1381 tgtacaccct aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc
     1441 tcctcgttcg accccgcctc gatcctccct ttatccagcc ctcactcctt ctctaggcgc
     1501 ccccatatgg ccatatgaga tcttatatgg ggcacccccg ccccttgtaa acttccctga
     1561 ccctgacatg acaagagtta ctaacagccc ctctctccaa gctcacttac aggctctcta
     1621 cttagtccag cacgaagtct ggagacctct ggcggcagcc taccaagaac aactggaccg
     1681 accggtggta cctcaccctt accgagtcgg cgacacagtg tgggtccgcc gacaccagac
     1741 taagaaccta gaacctcgct ggaaaggacc ttacacagtc ctgctgacca cccccaccgc
     1801 cctcaaagta gacggcatcg cagcttggat acacgccgcc cacgtgaagg ctgccgaccc
     1861 cgggggtgga ccatcctcta gactgccgga tctagctagt taattaagga tcccagtgtg
     1921 gtggtacggg gatccaccat gtacccatac gatgttccag attacgctaa gttccagtac
     1981 aaggaggacc atccctttga gtatcggaaa aaggaaggag aaaagatccg gaagaaatat
     2041 ccggacaggg tccccgtgat tgtagagaag gctccaaaag ccagggtgcc tgatctggac
     2101 aagaggaagt acctagtgcc ctctgacctt actgttggcc agttctactt cttaatccgg
     2161 aagagaatcc acctgagacc tgaggacgcc ttattcttct ttgtcaacaa caccatccct
     2221 cccaccagtg ctaccatggg ccaactgtat gaggacaatc atgaggaaga ctattttctg
     2281 tatgtggcct acagtgatga gagtgtctat gggaaatgaa ctcgagggcc ggcgcgccgc
     2341 ggccgctacg taaattccgc ccctctccct cccccccccc taacgttact ggccgaagcc
     2401 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     2461 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     2521 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     2581 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     2641 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     2701 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     2761 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     2821 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaacgtct
     2881 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aagcttgcca
     2941 caaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     3001 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     3061 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     3121 ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
     3181 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     3241 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     3301 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3361 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     3421 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     3481 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3541 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     3601 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3661 agtaaagcgg cccagccaca gtggtcgacg ataaaataaa agattttatt tagtctccag
     3721 aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca
     3781 ttttggaagg catggaaaaa tacataactg agaatagaga agttcagatc aaggtcagga
     3841 acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     3901 ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat atctgtggta
     3961 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtccagccct
     4021 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc
     4081 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct
     4141 ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc tccgattgac
     4201 tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg
     4261 tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt
     4321 tcattctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4381 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4441 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4501 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4561 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4621 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4681 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4741 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4801 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4861 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4921 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4981 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     5041 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5101 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5161 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5221 tcatgagatt atcaaaaagg atcttcacct agatcctttt gcggccggcc gcaaatcaat
     5281 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5341 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5401 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5461 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5521 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5581 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5641 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5701 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5761 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5821 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5881 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa
     5941 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6001 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6061 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6121 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6181 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6241 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttc
//
