LOCUS       sequence-233023-        9212 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of LIR double mutant NBR1 without its UBD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9212)
  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 9212)
  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 mCherry-NBR1(F562A/Y731A/Delta-UBD)
FEATURES             Location/Qualifiers
     source          1..9212
                     /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             2113..2820
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2258..2276)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2526..2545
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     LTR             5759..6228
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6358..6375)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6529..7117)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6609..6628)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7281..8072,8073..8141))
                     /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     7904..7923
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8142..8246)
                     /gene="bla"
                     /label=AmpR promoter
     mobile_element  8391..9158
                     /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 gaggataaca tggccatcat caaggagttc
     2161 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     2221 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     2281 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     2341 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     2401 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     2461 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     2521 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     2581 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     2641 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     2701 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     2761 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     2821 tccggactca gatctcgagc tcaagcttgg gaaccacagg ttactctaaa tgtgactttt
     2881 aaaaatgaaa ttcaaagctt tctggtttct gatccagaaa atacaacttg ggctgatatc
     2941 gaagctatgg taaaagtttc atttgatctg aatactattc aaataaaata cctggatgag
     3001 gaaaatgaag aggtatccat caacagtcaa ggagaatatg aagaagcgct taagatggca
     3061 gttaaacagg gaaaccaact gcagatgcaa gtccacgaag ggcaccatgt cgttgatgaa
     3121 gccccacccc cagttgtagg agcaaaacga ctagctgcca gggcagggaa gaagccactt
     3181 gcacattact cttcactggt gagagtcttg ggatcagaca tgaagacccc agaggatcct
     3241 gcagtgcagt cgtttccact tgttccatgt gacacagacc agcctcaaga caagccccca
     3301 gactggttca caagctacct ggagacgttc agagaacaag tggttaacga aacggttgag
     3361 aagcttgaac agaaattaca tgaaaagctt gtcctccaga acccatcctt gggttcttgt
     3421 ccctcagaag tctcaatgcc tacttcagaa gaaacattgt ttttgccaga aaaccagttc
     3481 agctggcata ttgcttgcaa caactgccaa agaaggattg ttggtgtccg ctaccagtgt
     3541 agcctatgcc catcctacaa tatctgtgaa gattgtgaag cagggccata tggccatgac
     3601 actaaccacg tcctgctgaa gttgcggaga cctgttgtgg gctcctctga accgttctgt
     3661 cactcaaagt actctactcc tcgtcttcct gctgctctgg aacaagtcag gctccagaaa
     3721 caggttgata agaactttct taaagcagaa aagcaaaggt tgcgagctga gaagaaacaa
     3781 cgtaaagcag aggtcaagga acttaaaaag cagcttaaac tccataggaa aattcacctg
     3841 tggaattcaa tccatggact ccagagcccc aagtctcctt taggccgacc tgagagcttg
     3901 ctccagtcta ataccctgat gctccctttg cagccctgta cctccgttat gccaatgctc
     3961 agtgcagcat ttgtggatga gaatttgcct gatgggactc accttcagcc aggaaccaag
     4021 tttatcaaac actggaggat gaaaaataca ggaaatgtaa agtggagtgc agacacaaag
     4081 ctcaagttca tgtggggaaa cctgactttg gcttccacag aaaagaagga tgttttggtt
     4141 ccctgcctca aggccggcca tgtgggagtt gtatctgtgg agttcattgc cccagccttg
     4201 gagggaacgt atacttccca ttggcgtctt tctcacaaag gccagcaatt tgggcctcgg
     4261 gtctggtgca gtatcatagt agatcctttc ccctccgaag agagccctga taacattgaa
     4321 aagggcatga tcagctcaag caaaactgat gatctcacct gccagcaaga ggaaactttt
     4381 cttctggcta aagaagaaag acagcttggt gaagtgactg agcagacaga agggacagca
     4441 gcctgcatcc cacagaaggc aaaaaatgtt gccagtgaga gggagctcta catcccatct
     4501 gtggatcttc tgactgccca ggacctgctg tccgcagagc tgttggatat aaacattgtt
     4561 caagagttgg agagagtgcc ccacaacacc cctgtggatg tgactccctg catgtctcct
     4621 ctgccacatg acagtccttt aatagagaag ccaggcttgg ggcagataga ggaagagaat
     4681 gaaggggcag gatttaaagc acttcctgat tctatggtgt cagtaaagag gaaggctgag
     4741 aacattgctt ctgtggagga agcagaagaa gacctgagtg ggacccagtt tgtgtgtgag
     4801 acagtaatcc gatcccttac cttggatgct gccccagacc acaaccctcc ttgcagacag
     4861 aagtccttgc agatgacatt tgccttgcct gaaggaccac ttggaaatga gaaggaggag
     4921 attatccata tcgctgagga agaagctgtc atggaggagg aggaggatga ggaggatgag
     4981 gaggaggagg atgagctcaa agatgaagtt caaagtcagt cctctgcttc ctcagaggat
     5041 gccatcatca tcctgcctga gtgctttgat accagccgcc ccctggggga ttctatgtac
     5101 agctctgcgc tctcacagcc aggcctggag cgaggtgctg aaggcaagcc tggggttgag
     5161 gctgggcagg aaccagctga ggctggggaa agactccctg gaggggagaa ccagccacag
     5221 gagcacagca taagtgacat cctcacgacc tcacagactc tggaaacagt gcccctaatc
     5281 ccagaggtag tggagcttcc accgtcactg cccaggagct ctccttgtgt acatcatcat
     5341 ggttccccag gagtggattt accagttacc ataccagaag tttcttcagt ccctgatcag
     5401 atcagaggag agcccagagg ctcatcagga cttgtaaaca gcagacagaa gagctatgac
     5461 cactcaaggc accatcatgg gagcagcatt gctggaggac tggtgaaggg ggctttgtct
     5521 gttgctgcct ctgcatacaa ggccctgttt gctgggccac catgaggatc caccggatct
     5581 agataactga tcacgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg
     5641 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg
     5701 aaaaatacat aactgagaat agagaagttc agatcaaggt caggaacaga tggaacagct
     5761 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     5821 acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     5881 ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag
     5941 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     6001 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     6061 aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta
     6121 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
     6181 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt
     6241 atcaaaatta atttggtttt ttttcttaag tatttacatt aaatggccat agttgcatta
     6301 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6361 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6421 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6481 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6541 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6601 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6661 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6721 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6781 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6841 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6901 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6961 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7021 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7081 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7141 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7201 aaaaaggatc ttcacctaga tccttttgcg gccggccgca aatcaatcta aagtatatat
     7261 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7321 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7381 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7441 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7501 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7561 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7621 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7681 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7741 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7801 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7861 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7921 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7981 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8041 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8101 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8161 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8221 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc agctttgctc
     8281 ttaggagttt cctaatacat cccaaactca aatatataaa gcatttgact tgttctatgc
     8341 cctagggggc ggggggaagc taagccagct ttttttaaca tttaaaatgt ggtaatgact
     8401 ccaacttatt gatagtgttt tatgttcaga taatgcccga tgactttgtc atgcagctcc
     8461 accgattttg agaacgacag cgacttccgt cccagccgtg ccaggtgctg cctcagattc
     8521 aggttatgcc gctcaattcg ctgcgtatat cgcttgctga ttacgtgcag ctttcccttc
     8581 aggcgggatt catacagcgg ccagccatcc gtcatccata tcaccacgtc aaagggtgac
     8641 agcaggctca taagacgccc cagcgtcgcc atagtgcgtt caccgaatac gtgcgcaaca
     8701 accgtcttcc ggagactgtc atacgcgtaa aacagccagc gctggcgcga tttagccccg
     8761 acatagcccc actgttcgtc catttccgcg cagacgatga cgtcactgcc cggctgtatg
     8821 cgcgaggtta ccgactgcgg cctgagtttt ttaagtgacg taaaatcgtg ttgaggccaa
     8881 cgcccataat gcgggctgtt gcccggcatc caacgccatt catggccata tcaatgattt
     8941 tctggtgcgt accgggttga gaagcggtgt aagtgaactg cagttgccat gttttacggc
     9001 agtgagagca gagatagcgc tgatgtccgg cggtgctttt gccgttacgc accaccccgt
     9061 cagtagctga acaggaggga cagctgatag aaacagaagc cactggagca cctcaaaaac
     9121 accatcatac actaaatcag taagttggca gcatcacctt aaaatgttaa ttccatttta
     9181 aatgcacaga tgtttttatt tcataagggt tt
//
