LOCUS       sequence-233077-        7835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of LIR/UBD double mutant NDP52 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7835)
  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 7835)
  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-NDP52(V136S/C443K)
FEATURES             Location/Qualifiers
     source          1..7835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             353..822
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    887..1244
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1309..1725
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1655..1677
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1693..1709
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1735..2109
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     2047..2063
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      2112..2121
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2118..2825
                     /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(2263..2281)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2531..2550
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     LTR             4387..4856
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4986..5003)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5157..5745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5237..5256)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5909..6700,6701..6769))
                     /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     6532..6551
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6770..6874)
                     /gene="bla"
                     /label=AmpR promoter
     mobile_element  7019..7786
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
ORIGIN
        1 ggtttcaatg tgcatgaatg ctgcaatatt cctgttacca aagctagtat aaataaaaat
       61 agataaacgt ggaaattact tagagtttct gtcattaacg tttccttcct cagttgacaa
      121 cataaatgcg ctgctgagaa gccagtttgc atctgtcagg atcaatttcc cattatgcca
      181 gtcatattaa ttactagtca attagttgat ttttattttt gacatataca tgtgaaagac
      241 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat
      301 acataactga gaatagaaaa gttcagatca aggtcaggaa cagatggaac agctgaatat
      361 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
      421 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
      481 gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat
      541 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
      601 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
      661 ccacaacccc tcactcggcg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg
      721 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
      781 tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg ctcgtccggg
      841 atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc tggccagcaa
      901 cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc ctgcgtcggt
      961 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttcggaa
     1021 cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt tgtggcccga
     1081 cctgagtcca aaaatcccga tcgttttgga ctctttggtg cacccccctt agaggaggga
     1141 tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg
     1201 ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat cgttctgtgt
     1261 tgtctctgtc tgactgtgtt tctgtatttg tctgaaaata tgggcccggg ccagactgtt
     1321 accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac
     1381 cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc
     1441 tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag
     1501 atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc
     1561 tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct
     1621 ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg
     1681 cctcgatcct ccctttatcc agccctcact ccttctctag gcgcccccat atggccatat
     1741 gagatcttat atggggcacc cccgcccctt gtaaacttcc ctgaccctga catgacaaga
     1801 gttactaaca gcccctctct ccaagctcac ttacaggctc tctacttagt ccagcacgaa
     1861 gtctggagac ctctggcggc agcctaccaa gaacaactgg accgaccggt ggtacctcac
     1921 ccttaccgag tcggcgacac agtgtgggtc cgccgacacc agactaagaa cctagaacct
     1981 cgctggaaag gaccttacac agtcctgctg accaccccca ccgccctcaa agtagacggc
     2041 atcgcagctt ggatacacgc cgcccacgtg aaggctgccg accccggggg tggaccatcc
     2101 tctagactgc cgccaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
     2161 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
     2221 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     2281 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
     2341 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
     2401 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
     2461 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
     2521 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
     2581 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
     2641 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
     2701 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
     2761 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
     2821 acaagtccgg actcagatct cgagctcaag cttgggagga gaccatcaaa gatcccccca
     2881 catcagctgt cttgctggat cactgtcatt tctctcaggt catctttaac agtgtggaga
     2941 agttctacat ccctggaggg gacgtcacat gtcattatac cttcacccag catttcatcc
     3001 ctcgtcgaaa ggattggatt ggcatcttta gagtggggtg gaagacaacc cgtgagtatt
     3061 acaccttcat gtgggttact ttgcccattg acctaaacaa caaatcagct aaacagcagg
     3121 aagtccaatt caaagcttac tacctgccca aggatgatga gtattaccag ttctgctatg
     3181 tggatgagga tggtgtggtc cggggagcaa gtattccttt ccaattccgt ccagaaaatg
     3241 aggaagacat cctggttagt accactcagg gagaggtgga agagattgag cagcacaaca
     3301 aggagctttg caaagaaaac caggagctga aggacagctg tatcagcctc cagaagcaga
     3361 actcagacat gcaggctgag ctccaaaaga agcaggagga gctagaaacc ctacagagca
     3421 tcaataagaa gttggaactg aaagtgaaag aacagaagga ctattgggag acagagctgc
     3481 ttcaactgaa agaacaaaac cagaagatgt cctcagaaaa tgagaagatg ggaatcagag
     3541 tggatcagct tcaggcccag ctgtcaactc aagagaaaga aatggagaag cttgttcagg
     3601 gagatcaaga taagacagag cagttagagc agctgaaaaa ggaaaatgac cacctctttc
     3661 tcagtttagc tgaacagagg aaggaccaga agaagctcga gcagacagtg gagcaaatga
     3721 agcagaatga aactactgca atgaagaaac aacaggaatt aatggatgaa aactttgacc
     3781 tgtcaaaaag actgagtgag aacgaaatta tatgtaatgc tctgcagaga cagaaagaga
     3841 gattggaagg agaaaatgat cttttgaaga gggagaacag cagattgctc agttacatgg
     3901 gtctggattt taattctttg ccgtatcaag tacctacttc agatgaagga ggcgcaagac
     3961 aaaatccagg acttgcctat ggaaacccat attctggtat ccaagaaagt tcttccccca
     4021 gcccgctctc catcaagaaa tgccctatct gcaaagcaga tgatatttgt gatcacacct
     4081 tggagcaaca gcagatgcag cccctttgtt tcaattgtcc aatttgtgac aagatcttcc
     4141 cagctacaga gaagcagatc tttgaagacc acgtgttcaa acactctctc tgaggatcca
     4201 ccggatctag ataactgatc acgataaaat aaaagatttt atttagtctc cagaaaaagg
     4261 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     4321 aggcatggaa aaatacataa ctgagaatag agaagttcag atcaaggtca ggaacagatg
     4381 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     4441 ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
     4501 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt
     4561 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
     4621 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
     4681 ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg
     4741 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
     4801 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcacatg
     4861 cagcatgtat caaaattaat ttggtttttt ttcttaagta tttacattaa atggccatag
     4921 ttgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4981 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5041 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5101 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5161 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5221 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5281 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5341 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5401 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5461 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5521 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5581 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5641 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5701 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5761 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5821 agattatcaa aaaggatctt cacctagatc cttttgcggc cggccgcaaa tcaatctaaa
     5881 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5941 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6001 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6061 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6121 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6181 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6241 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6301 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6361 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6421 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6481 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6541 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6601 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6661 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6721 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6781 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6841 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccag
     6901 ctttgctctt aggagtttcc taatacatcc caaactcaaa tatataaagc atttgacttg
     6961 ttctatgccc tagggggcgg ggggaagcta agccagcttt ttttaacatt taaaatgtgg
     7021 taatgactcc aacttattga tagtgtttta tgttcagata atgcccgatg actttgtcat
     7081 gcagctccac cgattttgag aacgacagcg acttccgtcc cagccgtgcc aggtgctgcc
     7141 tcagattcag gttatgccgc tcaattcgct gcgtatatcg cttgctgatt acgtgcagct
     7201 ttcccttcag gcgggattca tacagcggcc agccatccgt catccatatc accacgtcaa
     7261 agggtgacag caggctcata agacgcccca gcgtcgccat agtgcgttca ccgaatacgt
     7321 gcgcaacaac cgtcttccgg agactgtcat acgcgtaaaa cagccagcgc tggcgcgatt
     7381 tagccccgac atagccccac tgttcgtcca tttccgcgca gacgatgacg tcactgcccg
     7441 gctgtatgcg cgaggttacc gactgcggcc tgagtttttt aagtgacgta aaatcgtgtt
     7501 gaggccaacg cccataatgc gggctgttgc ccggcatcca acgccattca tggccatatc
     7561 aatgattttc tggtgcgtac cgggttgaga agcggtgtaa gtgaactgca gttgccatgt
     7621 tttacggcag tgagagcaga gatagcgctg atgtccggcg gtgcttttgc cgttacgcac
     7681 caccccgtca gtagctgaac aggagggaca gctgatagaa acagaagcca ctggagcacc
     7741 tcaaaaacac catcatacac taaatcagta agttggcagc atcaccttaa aatgttaatt
     7801 ccattttaaa tgcacagatg tttttatttc ataag
//
