LOCUS       sequence-233075-        8060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of iRFP-tagged LIR/UBD double mutant NDP52 
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8060)
  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 8060)
  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 iRFP670-NDP52(V136S/C443K)
FEATURES             Location/Qualifiers
     source          1..8060
                     /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..3050
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     LTR             4612..5081
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5211..5228)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5382..5970)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5462..5481)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6134..6925,6926..6994))
                     /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     6757..6776
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6995..7099)
                     /gene="bla"
                     /label=AmpR promoter
     mobile_element  7244..8011
                     /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 gcgcgtaagg tcgatctcac ctcctgcgat cgcgagccga
     2161 tccacatccc cggcagcatt cagccgtgcg gctgcctgct agcctgcgac gcgcaggcgg
     2221 tgcggatcac gcgcattacg gaaaatgccg gcgcgttctt tggacgcgaa actccgcggg
     2281 tcggtgagct actcgccgat tacttcggcg agaccgaagc ccatgcgctg cgcaacgcac
     2341 tggcgcagtc ctccgatcca aagcgaccgg cgctgatctt cggttggcgc gacggcctga
     2401 ccggccgcac cttcgacatc tcactgcatc gccatgacgg tacatcgatc atcgagttcg
     2461 agcctgcggc ggccgaacag gccgacaatc cgctgcggct gacgcggcag atcatcgcgc
     2521 gcaccaaaga actgaagtcg ctcgaagaga tggccgcacg ggtgccgcgc tatctgcagg
     2581 cgatgctcgg ctatcaccgc gtgatgttgt accgcttcgc ggacgacggc tccgggatgg
     2641 tgatcggcga ggcgaagcgc agcgacctcg agagctttct cggtcagcac tttccggcgt
     2701 cgctggtccc gcagcaggcg cggctactgt acttgaagaa cgcgatccgc gtggtctcgg
     2761 attcgcgcgg catcagcagc cggatcgtgc ccgagcacga cgcctccggc gccgcgctcg
     2821 atctgtcgtt cgcgcacctg cgcagcatct cgccctgcca tctcgaattt ctgcggaaca
     2881 tgggcgtcag cgcctcgatg tcgctgtcga tcatcattga cggcacgcta tggggattga
     2941 tcatctgtca tcattacgag ccgcgtgccg tgccgatggc gcagcgcgtc gcggccgaaa
     3001 tgttcgccga cttcttatcg ctgcacttca ccgccgccca ccaccaacgc tccggactca
     3061 gatctcgagc tcaagcttgg gaggagacca tcaaagatcc ccccacatca gctgtcttgc
     3121 tggatcactg tcatttctct caggtcatct ttaacagtgt ggagaagttc tacatccctg
     3181 gaggggacgt cacatgtcat tataccttca cccagcattt catccctcgt cgaaaggatt
     3241 ggattggcat ctttagagtg gggtggaaga caacccgtga gtattacacc ttcatgtggg
     3301 ttactttgcc cattgaccta aacaacaaat cagctaaaca gcaggaagtc caattcaaag
     3361 cttactacct gcccaaggat gatgagtatt accagttctg ctatgtggat gaggatggtg
     3421 tggtccgggg agcaagtatt cctttccaat tccgtccaga aaatgaggaa gacatcctgg
     3481 ttagtaccac tcagggagag gtggaagaga ttgagcagca caacaaggag ctttgcaaag
     3541 aaaaccagga gctgaaggac agctgtatca gcctccagaa gcagaactca gacatgcagg
     3601 ctgagctcca aaagaagcag gaggagctag aaaccctaca gagcatcaat aagaagttgg
     3661 aactgaaagt gaaagaacag aaggactatt gggagacaga gctgcttcaa ctgaaagaac
     3721 aaaaccagaa gatgtcctca gaaaatgaga agatgggaat cagagtggat cagcttcagg
     3781 cccagctgtc aactcaagag aaagaaatgg agaagcttgt tcagggagat caagataaga
     3841 cagagcagtt agagcagctg aaaaaggaaa atgaccacct ctttctcagt ttagctgaac
     3901 agaggaagga ccagaagaag ctcgagcaga cagtggagca aatgaagcag aatgaaacta
     3961 ctgcaatgaa gaaacaacag gaattaatgg atgaaaactt tgacctgtca aaaagactga
     4021 gtgagaacga aattatatgt aatgctctgc agagacagaa agagagattg gaaggagaaa
     4081 atgatctttt gaagagggag aacagcagat tgctcagtta catgggtctg gattttaatt
     4141 ctttgccgta tcaagtacct acttcagatg aaggaggcgc aagacaaaat ccaggacttg
     4201 cctatggaaa cccatattct ggtatccaag aaagttcttc ccccagcccg ctctccatca
     4261 agaaatgccc tatctgcaaa gcagatgata tttgtgatca caccttggag caacagcaga
     4321 tgcagcccct ttgtttcaat tgtccaattt gtgacaagat cttcccagct acagagaagc
     4381 agatctttga agaccacgtg ttcaaacact ctctctgagg atccaccgga tctagataac
     4441 tgatcacgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     4501 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata
     4561 cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg
     4621 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     4681 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     4741 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc
     4801 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     4861 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     4921 cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
     4981 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     5041 ctcctctgag tgattgacta cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa
     5101 ttaatttggt tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc
     5161 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5221 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5281 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5341 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5401 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5461 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5521 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5581 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5641 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5701 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5761 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5821 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5881 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5941 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6001 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6061 atcttcacct agatcctttt gcggccggcc gcaaatcaat ctaaagtata tatgagtaaa
     6121 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6181 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6241 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6301 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6361 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6421 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6481 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6541 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6601 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6661 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6721 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6781 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6841 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6901 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6961 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7021 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7081 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc accagctttg ctcttaggag
     7141 tttcctaata catcccaaac tcaaatatat aaagcatttg acttgttcta tgccctaggg
     7201 ggcgggggga agctaagcca gcttttttta acatttaaaa tgtggtaatg actccaactt
     7261 attgatagtg ttttatgttc agataatgcc cgatgacttt gtcatgcagc tccaccgatt
     7321 ttgagaacga cagcgacttc cgtcccagcc gtgccaggtg ctgcctcaga ttcaggttat
     7381 gccgctcaat tcgctgcgta tatcgcttgc tgattacgtg cagctttccc ttcaggcggg
     7441 attcatacag cggccagcca tccgtcatcc atatcaccac gtcaaagggt gacagcaggc
     7501 tcataagacg ccccagcgtc gccatagtgc gttcaccgaa tacgtgcgca acaaccgtct
     7561 tccggagact gtcatacgcg taaaacagcc agcgctggcg cgatttagcc ccgacatagc
     7621 cccactgttc gtccatttcc gcgcagacga tgacgtcact gcccggctgt atgcgcgagg
     7681 ttaccgactg cggcctgagt tttttaagtg acgtaaaatc gtgttgaggc caacgcccat
     7741 aatgcgggct gttgcccggc atccaacgcc attcatggcc atatcaatga ttttctggtg
     7801 cgtaccgggt tgagaagcgg tgtaagtgaa ctgcagttgc catgttttac ggcagtgaga
     7861 gcagagatag cgctgatgtc cggcggtgct tttgccgtta cgcaccaccc cgtcagtagc
     7921 tgaacaggag ggacagctga tagaaacaga agccactgga gcacctcaaa aacaccatca
     7981 tacactaaat cagtaagttg gcagcatcac cttaaaatgt taattccatt ttaaatgcac
     8041 agatgttttt atttcataag
//
