LOCUS       sequence-372301-        7859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of NDP52 tagged with mEGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7859)
  AUTHORS   Nguyen TN, Sawa-Makarska J, Khuu G, Lam WK, Adriaenssens E, 
            Fracchiolla D, Shoebridge S, Bernklau D, Padman BS, Skulsuppaisarn 
            M, Lindblom RSJ, Martens S, Lazarou M
  TITLE     Unconventional initiation of PINK1/Parkin mitophagy by Optineurin.
  JOURNAL   Mol Cell. 2023 May 18;83(10):1693-1709.e9. doi: 
            10.1016/j.molcel.2023.04.021.
  PUBMED    37207627
REFERENCE   2  (bases 1 to 7859)
  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"; doi: 
            "10.1016/j.molcel.2023.04"; journalName: "Mol Cell"; date: 
            "2023-05-18- 18"; volume: "83"; issue: "10"; pages: "1693-1709.e9"
COMMENT     Sequence Label: pBMN mEGFP-NDP52
FEATURES             Location/Qualifiers
     source          1..7859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  188..955
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     LTR             1357..1826
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    1891..2248
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             2313..2729
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2659..2681
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2697..2713
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    2739..3113
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     3051..3067
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      3116..3125
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3122..3124,3125..3127,3128..3838)
                     /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(3167..3188)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3428..3447)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3775..3796
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5415..5884
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6014..6031)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6185..6773)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6265..6284)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6937..7728,7729..7797))
                     /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     7560..7579
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7798..7859,1..43))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
       61 tgccaccagc tttgctctta ggagtttcct aatacatccc aaactcaaat atataaagca
      121 tttgacttgt tctatgccct agggggcggg gggaagctaa gccagctttt tttaacattt
      181 aaaatgtggt aatgactcca acttattgat agtgttttat gttcagataa tgcccgatga
      241 ctttgtcatg cagctccacc gattttgaga acgacagcga cttccgtccc agccgtgcca
      301 ggtgctgcct cagattcagg ttatgccgct caattcgctg cgtatatcgc ttgctgatta
      361 cgtgcagctt tcccttcagg cgggattcat acagcggcca gccatccgtc atccatatca
      421 ccacgtcaaa gggtgacagc aggctcataa gacgccccag cgtcgccata gtgcgttcac
      481 cgaatacgtg cgcaacaacc gtcttccgga gactgtcata cgcgtaaaac agccagcgct
      541 ggcgcgattt agccccgaca tagccccact gttcgtccat ttccgcgcag acgatgacgt
      601 cactgcccgg ctgtatgcgc gaggttaccg actgcggcct gagtttttta agtgacgtaa
      661 aatcgtgttg aggccaacgc ccataatgcg ggctgttgcc cggcatccaa cgccattcat
      721 ggccatatca atgattttct ggtgcgtacc gggttgagaa gcggtgtaag tgaactgcag
      781 ttgccatgtt ttacggcagt gagagcagag atagcgctga tgtccggcgg tgcttttgcc
      841 gttacgcacc accccgtcag tagctgaaca ggagggacag ctgatagaaa cagaagccac
      901 tggagcacct caaaaacacc atcatacact aaatcagtaa gttggcagca tcaccttaaa
      961 atgttaattc cattttaaat gcacagatgt ttttatttca taagggtttc aatgtgcatg
     1021 aatgctgcaa tattcctgtt accaaagcta gtataaataa aaatagataa acgtggaaat
     1081 tacttagagt ttctgtcatt aacgtttcct tcctcagttg acaacataaa tgcgctgctg
     1141 agaagccagt ttgcatctgt caggatcaat ttcccattat gccagtcata ttaattacta
     1201 gtcaattagt tgatttttat ttttgacata tacatgtgaa agaccccacc tgtaggtttg
     1261 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag
     1321 aaaagttcag atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat
     1381 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg
     1441 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     1501 gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt
     1561 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
     1621 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
     1681 ggcgcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc
     1741 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
     1801 gactacccgt cagcgggggt ctttcatttg ggggctcgtc cgggatcggg agacccctgc
     1861 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
     1921 cgattgtcta gtgtctatga ctgattttat gcgcctgcgt cggtactagt tagctaacta
     1981 gctctgtatc tggcggaccc gtggtggaac tgacgagttc ggaacacccg gccgcaaccc
     2041 tgggagacgt cccagggact tcgggggccg tttttgtggc ccgacctgag tccaaaaatc
     2101 ccgatcgttt tggactcttt ggtgcacccc ccttagagga gggatatgtg gttctggtag
     2161 gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttgggacc
     2221 gaagccgcgc cgcgcgtctt gtctgctgca gcatcgttct gtgttgtctc tgtctgactg
     2281 tgtttctgta tttgtctgaa aatatgggcc cgggccagac tgttaccact cccttaagtt
     2341 tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca
     2401 agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc
     2461 cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac
     2521 ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt
     2581 ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc
     2641 catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt
     2701 atccagccct cactccttct ctaggcgccc ccatatggcc atatgagatc ttatatgggg
     2761 cacccccgcc ccttgtaaac ttccctgacc ctgacatgac aagagttact aacagcccct
     2821 ctctccaagc tcacttacag gctctctact tagtccagca cgaagtctgg agacctctgg
     2881 cggcagccta ccaagaacaa ctggaccgac cggtggtacc tcacccttac cgagtcggcg
     2941 acacagtgtg ggtccgccga caccagacta agaacctaga acctcgctgg aaaggacctt
     3001 acacagtcct gctgaccacc cccaccgccc tcaaagtaga cggcatcgca gcttggatac
     3061 acgccgccca cgtgaaggct gccgaccccg ggggtggacc atcctctaga ctgccgccac
     3121 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     3181 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     3241 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     3301 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     3361 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     3421 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     3481 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     3541 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     3601 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     3661 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     3721 ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     3781 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc
     3841 cggactcaga tctcgagctc aagcttcgaa ttctgcagtc gacgaggaga ccatcaaaga
     3901 tccccccaca tcagctgtct tgctggatca ctgtcatttc tctcaggtca tctttaacag
     3961 tgtggagaag ttctacatcc ctggagggga cgtcacatgt cattatacct tcacccagca
     4021 tttcatccct cgtcgaaagg attggattgg catctttaga gtggggtgga agacaacccg
     4081 tgagtattac accttcatgt gggttacttt gcccattgac ctaaacaaca aatcagctaa
     4141 acagcaggaa gtccaattca aagcttacta cctgcccaag gatgatgagt attaccagtt
     4201 ctgctatgtg gatgaggatg gtgtggtccg gggagcaagt attcctttcc aattccgtcc
     4261 agaaaatgag gaagacatcc tggttgttac cactcaggga gaggtggaag agattgagca
     4321 gcacaacaag gagctttgca aagaaaacca ggagctgaag gacagctgta tcagcctcca
     4381 gaagcagaac tcagacatgc aggctgagct ccaaaagaag caggaggagc tagaaaccct
     4441 acagagcatc aataagaagt tggaactgaa agtgaaagaa cagaaggact attgggagac
     4501 agagctgctt caactgaaag aacaaaacca gaagatgtcc tcagaaaatg agaagatggg
     4561 aatcagagtg gatcagcttc aggcccagct gtcaactcaa gagaaagaaa tggagaagct
     4621 tgttcaggga gatcaagata agacagagca gttagagcag ctgaaaaagg aaaatgacca
     4681 cctctttctc agtttagctg aacagaggaa ggaccagaag aagctcgagc agacagtgga
     4741 gcaaatgaag cagaatgaaa ctactgcaat gaagaaacaa caggaattaa tggatgaaaa
     4801 ctttgacctg tcaaaaagac tgagtgagaa cgaaattata tgtaatgctc tgcagagaca
     4861 gaaagagaga ttggaaggag aaaatgatct tttgaagagg gagaacagca gattgctcag
     4921 ttacatgggt ctggatttta attctttgcc gtatcaagta cctacttcag atgaaggagg
     4981 cgcaagacaa aatccaggac ttgcctatgg aaacccatat tctggtatcc aagaaagttc
     5041 ttcccccagc ccgctctcca tcaagaaatg ccctatctgc aaagcagatg atatttgtga
     5101 tcacaccttg gagcaacagc agatgcagcc cctttgtttc aattgtccaa tttgtgacaa
     5161 gatcttccca gctacagaga agcagatctt tgaagaccac gtgttctgcc actctctctg
     5221 aggatccacc ggatctagat aactgatcac gataaaataa aagattttat ttagtctcca
     5281 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     5341 attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg
     5401 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     5461 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     5521 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc
     5581 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     5641 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     5701 tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga
     5761 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     5821 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     5881 ttcacatgca gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat
     5941 ggccatagtt gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6001 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6061 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6121 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6181 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6241 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6301 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6361 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6421 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6481 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6541 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6601 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6661 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6721 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6781 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6841 tggtcatgag attatcaaaa aggatcttca cctagatcct tttgcggccg gccgcaaatc
     6901 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6961 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7021 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7081 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7141 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7201 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7261 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7321 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7381 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7441 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7501 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7561 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7621 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7681 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7741 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7801 cttccttttt caatattatt gaagcattta tcagggttat tatctcatga gcggataca
//
