LOCUS       sequence-235957-        8234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of LIR mutant OPTN .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8234)
  AUTHORS   Padman BS, Nguyen TN, Uoselis L, Skulsuppaisarn M, Nguyen LK, 
            Lazarou M
  TITLE     LC3/GABARAPs drive ubiquitin-independent recruitment of Optineurin 
            and NDP52 to amplify mitophagy.
  JOURNAL   Nat Commun. 2019 Jan 24;10(1):408. doi: 10.1038/s41467-019-08335-6.
  PUBMED    30679426
REFERENCE   2  (bases 1 to 8234)
  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: "Nat 
            Commun."; date: "2019-01-24"; volume: "10(1):408. doi"; pages: " 
            10.1038/s41467-019-08335-6"
COMMENT     Sequence Label: pBMN mEGFP-OPTN(F178A)
FEATURES             Location/Qualifiers
     source          1..8234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             348..817
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    882..1239
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1304..1720
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1650..1672
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1688..1704
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1730..2104
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     2042..2058
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      2107..2116
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2113..2115,2116..2118,2119..2829)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2158..2179)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2419..2438)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2766..2787
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3171..3182
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4781..5250
                     /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"
     CDS             complement(join(6303..7094,7095..7163))
                     /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     6926..6945
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7164..7268)
                     /gene="bla"
                     /label=AmpR promoter
     mobile_element  7413..8180
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
ORIGIN
        1 caatgtgcat gaatgctgca atattcctgt taccaaagct agtataaata aaaatagata
       61 aacgtggaaa ttacttagag tttctgtcat taacgtttcc ttcctcagtt gacaacataa
      121 atgcgctgct gagaagccag tttgcatctg tcaggatcaa tttcccatta tgccagtcat
      181 attaattact agtcaattag ttgattttta tttttgacat atacatgtga aagaccccac
      241 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
      301 actgagaata gaaaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
      361 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
      421 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
      481 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
      541 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
      601 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
      661 acccctcact cggcgcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
      721 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
      781 tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg
      841 gagacccctg cccagggacc accgacccac caccgggagg taagctggcc agcaacttat
      901 ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg tcggtactag
      961 ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt cggaacaccc
     1021 ggccgcaacc ctgggagacg tcccagggac ttcgggggcc gtttttgtgg cccgacctga
     1081 gtccaaaaat cccgatcgtt ttggactctt tggtgcaccc cccttagagg agggatatgt
     1141 ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc
     1201 ggtttgggac cgaagccgcg ccgcgcgtct tgtctgctgc agcatcgttc tgtgttgtct
     1261 ctgtctgact gtgtttctgt atttgtctga aaatatgggc ccgggccaga ctgttaccac
     1321 tcccttaagt ttgaccttag gtcactggaa agatgtcgag cggatcgctc acaaccagtc
     1381 ggtagatgtc aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa
     1441 cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa
     1501 ggtcttttca cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga
     1561 agccttggct tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc
     1621 tcctcttcct ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg
     1681 atcctccctt tatccagccc tcactccttc tctaggcgcc cccatatggc catatgagat
     1741 cttatatggg gcacccccgc cccttgtaaa cttccctgac cctgacatga caagagttac
     1801 taacagcccc tctctccaag ctcacttaca ggctctctac ttagtccagc acgaagtctg
     1861 gagacctctg gcggcagcct accaagaaca actggaccga ccggtggtac ctcaccctta
     1921 ccgagtcggc gacacagtgt gggtccgccg acaccagact aagaacctag aacctcgctg
     1981 gaaaggacct tacacagtcc tgctgaccac ccccaccgcc ctcaaagtag acggcatcgc
     2041 agcttggata cacgccgccc acgtgaaggc tgccgacccc gggggtggac catcctctag
     2101 actgccgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2161 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2221 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2281 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     2341 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2401 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     2461 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2521 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     2581 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     2641 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2701 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc
     2761 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     2821 ctgtacaagt ccggactcag atctcgagct caagcttccc atcaacctct cagctgcctc
     2881 actgaaaagg aggacagccc cagtgaaagc acaggaaatg gaccccccca cctggcccac
     2941 ccaaacctgg acacgtttac cccggaggag ctgctgcagc agatgaaaga gctcctgacc
     3001 gagaaccacc agctgaaaga agccatgaag ctaaataatc aagccatgaa agggagattt
     3061 gaggagcttt cggcctggac agagaaacag aaggaagaac gccagttttt tgagatacag
     3121 agcaaagaag caaaagagcg tctaatggcc ttgagtcatg agaatgagaa attgaaggaa
     3181 gagcttggaa aactaaaagg gaaatcagaa aggtcatctg aggaccccac tgatgactcc
     3241 aggcttccca gggccgaagc ggagcaggaa aaggaccagc tcaggaccca ggtggtgagg
     3301 ctacaagcag agaaggcaga cctgttgggc atcgtgtctg aactgcagct caagctgaac
     3361 tccagcggct cctcagaaga ttccgctgtt gaaattagga tggctgaagg agaagcagaa
     3421 gggtcagtaa aagaaatcaa gcatagtcct gggcccacga gaacagtctc cactggcacg
     3481 gcattgtcta aatataggag cagatctgca gatggggcca agaattactt cgaacatgag
     3541 gagttaactg tgagccagct cctgctgtgc ctaagggaag ggaatcagaa ggtggagaga
     3601 cttgaagttg cactcaagga ggccaaagaa agagtttcag attttgaaaa gaaaacaagt
     3661 aatcgttctg agattgaaac ccagacagag gggagcacag agaaagagaa tgatgaagag
     3721 aaaggcccgg agactgttgg aagcgaagtg gaagcactga acctccaggt gacatctctg
     3781 tttaaggagc ttcaagaggc tcatacaaaa ctcagcgaag ctgagctaat gaagaagaga
     3841 cttcaagaaa agtgtcaggc ccttgaaagg aaaaattctg caattccatc agagttgaat
     3901 gaaaagcaag agcttgttta tactaacaaa aagttagagc tacaagtgga aagcatgcta
     3961 tcagaaatca aaatggaaca ggctaaaaca gaggatgaaa agtccaaatt aactgtgcta
     4021 cagatgacac acaacaagct tcttcaagaa cataataatg cattgaaaac aattgaggaa
     4081 ctaacaagaa aagagtcaga aaaagtggac agggcagtgc tgaaggaact gagtgaaaaa
     4141 ctggaactgg cagagaaggc tctggcttcc aaacagctgc aaatggatga aatgaagcaa
     4201 accattgcca agcaggaaga ggacctggaa accatgacca tcctcagggc tcagatggaa
     4261 gtttactgtt ctgattttca tgctgaaaga gcagcgagag agaaaattca tgaggaaaag
     4321 gagcaactgg cattgcagct ggcagttctg ctgaaagaga atgatgcttt cgaagacgga
     4381 ggcaggcagt ccttgatgga gatgcagagt cgtcatgggg cgagaacaag tgactctgac
     4441 cagcaggctt accttgttca aagaggagct gaggacaggg actggcggca acagcggaat
     4501 attccgattc attcctgccc caagtgtgga gaggttctgc ctgacataga cacgttacag
     4561 attcacgtga tggattgcat catttaagga tccaccggat ctagataact gatcacgata
     4621 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
     4681 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac ataactgaga
     4741 atagagaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg gccaaacagg
     4801 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa
     4861 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     4921 gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca
     4981 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     5041 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     5101 actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac
     5161 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
     5221 gattgactac ccgtcagcgg gggtctttca catgcagcat gtatcaaaat taatttggtt
     5281 ttttttctta agtatttaca ttaaatggcc atagttgcat 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 gaacagtatt
     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 cggccggccg caaatcaatc taaagtatat atgagtaaac ttggtctgac
     6301 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6361 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6421 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6481 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6541 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6601 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6661 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6721 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6781 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6841 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6901 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6961 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7021 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7081 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7141 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7201 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7261 gttccgcgca catttccccg aaaagtgcca ccagctttgc tcttaggagt ttcctaatac
     7321 atcccaaact caaatatata aagcatttga cttgttctat gccctagggg gcggggggaa
     7381 gctaagccag ctttttttaa catttaaaat gtggtaatga ctccaactta ttgatagtgt
     7441 tttatgttca gataatgccc gatgactttg tcatgcagct ccaccgattt tgagaacgac
     7501 agcgacttcc gtcccagccg tgccaggtgc tgcctcagat tcaggttatg ccgctcaatt
     7561 cgctgcgtat atcgcttgct gattacgtgc agctttccct tcaggcggga ttcatacagc
     7621 ggccagccat ccgtcatcca tatcaccacg tcaaagggtg acagcaggct cataagacgc
     7681 cccagcgtcg ccatagtgcg ttcaccgaat acgtgcgcaa caaccgtctt ccggagactg
     7741 tcatacgcgt aaaacagcca gcgctggcgc gatttagccc cgacatagcc ccactgttcg
     7801 tccatttccg cgcagacgat gacgtcactg cccggctgta tgcgcgaggt taccgactgc
     7861 ggcctgagtt ttttaagtga cgtaaaatcg tgttgaggcc aacgcccata atgcgggctg
     7921 ttgcccggca tccaacgcca ttcatggcca tatcaatgat tttctggtgc gtaccgggtt
     7981 gagaagcggt gtaagtgaac tgcagttgcc atgttttacg gcagtgagag cagagatagc
     8041 gctgatgtcc ggcggtgctt ttgccgttac gcaccacccc gtcagtagct gaacaggagg
     8101 gacagctgat agaaacagaa gccactggag cacctcaaaa acaccatcat acactaaatc
     8161 agtaagttgg cagcatcacc ttaaaatgtt aattccattt taaatgcaca gatgttttta
     8221 tttcataagg gttt
//
