LOCUS       sequence-372299-        8252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of OPTN tagged with mEGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8252)
  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 8252)
  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-OPTN
FEATURES             Location/Qualifiers
     source          1..8252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(64..168)
                     /gene="bla"
                     /label=AmpR promoter
     mobile_element  313..1080
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     LTR             1482..1951
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    2016..2373
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             2438..2854
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2784..2806
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2822..2838
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    2864..3238
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     3176..3192
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      3241..3250
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3247..3249,3250..3252,3253..3963)
                     /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(3292..3313)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3553..3572)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3900..3921
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4323..4334
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5933..6402
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6532..6549)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6703..7291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6783..6802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7455..8246,8247..8252,1..63))
                     /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     8078..8097
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
       61 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
      121 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
      181 aaaagtgcca ccagctttgc tcttaggagt ttcctaatac atcccaaact caaatatata
      241 aagcatttga cttgttctat gccctagggg gcggggggaa gctaagccag ctttttttaa
      301 catttaaaat gtggtaatga ctccaactta ttgatagtgt tttatgttca gataatgccc
      361 gatgactttg tcatgcagct ccaccgattt tgagaacgac agcgacttcc gtcccagccg
      421 tgccaggtgc tgcctcagat tcaggttatg ccgctcaatt cgctgcgtat atcgcttgct
      481 gattacgtgc agctttccct tcaggcggga ttcatacagc ggccagccat ccgtcatcca
      541 tatcaccacg tcaaagggtg acagcaggct cataagacgc cccagcgtcg ccatagtgcg
      601 ttcaccgaat acgtgcgcaa caaccgtctt ccggagactg tcatacgcgt aaaacagcca
      661 gcgctggcgc gatttagccc cgacatagcc ccactgttcg tccatttccg cgcagacgat
      721 gacgtcactg cccggctgta tgcgcgaggt taccgactgc ggcctgagtt ttttaagtga
      781 cgtaaaatcg tgttgaggcc aacgcccata atgcgggctg ttgcccggca tccaacgcca
      841 ttcatggcca tatcaatgat tttctggtgc gtaccgggtt gagaagcggt gtaagtgaac
      901 tgcagttgcc atgttttacg gcagtgagag cagagatagc gctgatgtcc ggcggtgctt
      961 ttgccgttac gcaccacccc gtcagtagct gaacaggagg gacagctgat agaaacagaa
     1021 gccactggag cacctcaaaa acaccatcat acactaaatc agtaagttgg cagcatcacc
     1081 ttaaaatgtt aattccattt taaatgcaca gatgttttta tttcataagg gtttcaatgt
     1141 gcatgaatgc tgcaatattc ctgttaccaa agctagtata aataaaaata gataaacgtg
     1201 gaaattactt agagtttctg tcattaacgt ttccttcctc agttgacaac ataaatgcgc
     1261 tgctgagaag ccagtttgca tctgtcagga tcaatttccc attatgccag tcatattaat
     1321 tactagtcaa ttagttgatt tttatttttg acatatacat gtgaaagacc ccacctgtag
     1381 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag
     1441 aatagaaaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag
     1501 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga
     1561 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     1621 agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc
     1681 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     1741 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     1801 cactcggcgc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     1861 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     1921 tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc
     1981 cctgcccagg gaccaccgac ccaccaccgg gaggtaagct ggccagcaac ttatctgtgt
     2041 ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta ctagttagct
     2101 aactagctct gtatctggcg gacccgtggt ggaactgacg agttcggaac acccggccgc
     2161 aaccctggga gacgtcccag ggacttcggg ggccgttttt gtggcccgac ctgagtccaa
     2221 aaatcccgat cgttttggac tctttggtgc acccccctta gaggagggat atgtggttct
     2281 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
     2341 ggaccgaagc cgcgccgcgc gtcttgtctg ctgcagcatc gttctgtgtt gtctctgtct
     2401 gactgtgttt ctgtatttgt ctgaaaatat gggcccgggc cagactgtta ccactccctt
     2461 aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga
     2521 tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg
     2581 atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt
     2641 ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt
     2701 ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct
     2761 tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc
     2821 cctttatcca gccctcactc cttctctagg cgcccccata tggccatatg agatcttata
     2881 tggggcaccc ccgccccttg taaacttccc tgaccctgac atgacaagag ttactaacag
     2941 cccctctctc caagctcact tacaggctct ctacttagtc cagcacgaag tctggagacc
     3001 tctggcggca gcctaccaag aacaactgga ccgaccggtg gtacctcacc cttaccgagt
     3061 cggcgacaca gtgtgggtcc gccgacacca gactaagaac ctagaacctc gctggaaagg
     3121 accttacaca gtcctgctga ccacccccac cgccctcaaa gtagacggca tcgcagcttg
     3181 gatacacgcc gcccacgtga aggctgccga ccccgggggt ggaccatcct ctagactgcc
     3241 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     3301 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     3361 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     3421 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     3481 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     3541 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     3601 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     3661 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     3721 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     3781 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3841 aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     3901 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     3961 aagtccggac tcagatctcg agctcaagct tcgaattctg cagtcgactc ccatcaacct
     4021 ctcagctgcc tcactgaaaa ggaggacagc cccagtgaaa gcacaggaaa tggacccccc
     4081 cacctggccc acccaaacct ggacacgttt accccggagg agctgctgca gcagatgaaa
     4141 gagctcctga ccgagaacca ccagctgaaa gaagccatga agctaaataa tcaagccatg
     4201 aaagggagat ttgaggagct ttcggcctgg acagagaaac agaaggaaga acgccagttt
     4261 tttgagatac agagcaaaga agcaaaagag cgtctaatgg ccttgagtca tgagaatgag
     4321 aaattgaagg aagagcttgg aaaactaaaa gggaaatcag aaaggtcatc tgaggacccc
     4381 actgatgact ccaggcttcc cagggccgaa gcggagcagg aaaaggacca gctcaggacc
     4441 caggtggtga ggctacaagc agagaaggca gacctgttgg gcatcgtgtc tgaactgcag
     4501 ctcaagctga actccagcgg ctcctcagaa gattcctttg ttgaaattag gatggctgaa
     4561 ggagaagcag aagggtcagt aaaagaaatc aagcatagtc ctgggcccac gagaacagtc
     4621 tccactggca cggcattgtc taaatatagg agcagatctg cagatggggc caagaattac
     4681 ttcgaacatg aggagttaac tgtgagccag ctcctgctgt gcctaaggga agggaatcag
     4741 aaggtggaga gacttgaagt tgcactcaag gaggccaaag aaagagtttc agattttgaa
     4801 aagaaaacaa gtaatcgttc tgagattgaa acccagacag aggggagcac agagaaagag
     4861 aatgatgaag agaaaggccc ggagactgtt ggaagcgaag tggaagcact gaacctccag
     4921 gtgacatctc tgtttaagga gcttcaagag gctcatacaa aactcagcga agctgagcta
     4981 atgaagaaga gacttcaaga aaagtgtcag gcccttgaaa ggaaaaattc tgcaattcca
     5041 tcagagttga atgaaaagca agagcttgtt tatactaaca aaaagttaga gctacaagtg
     5101 gaaagcatgc tatcagaaat caaaatggaa caggctaaaa cagaggatga aaagtccaaa
     5161 ttaactgtgc tacagatgac acacaacaag cttcttcaag aacataataa tgcattgaaa
     5221 acaattgagg aactaacaag aaaagagtca gaaaaagtgg acagggcagt gctgaaggaa
     5281 ctgagtgaaa aactggaact ggcagagaag gctctggctt ccaaacagct gcaaatggat
     5341 gaaatgaagc aaaccattgc caagcaggaa gaggacctgg aaaccatgac catcctcagg
     5401 gctcagatgg aagtttactg ttctgatttt catgctgaaa gagcagcgag agagaaaatt
     5461 catgaggaaa aggagcaact ggcattgcag ctggcagttc tgctgaaaga gaatgatgct
     5521 ttcgaagacg gaggcaggca gtccttgatg gagatgcaga gtcgtcatgg ggcgagaaca
     5581 agtgactctg accagcaggc ttaccttgtt caaagaggag ctgaggacag ggactggcgg
     5641 caacagcgga atattccgat tcattcctgc cccaagtgtg gagaggttct gcctgacata
     5701 gacacgttac agattcacgt gatggattgc atcatttaag gatccaccgg atctagataa
     5761 ctgatcacga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     5821 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat
     5881 acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac agctgaatat
     5941 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     6001 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     6061 gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat
     6121 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
     6181 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
     6241 ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg
     6301 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
     6361 tctcctctga gtgattgact acccgtcagc gggggtcttt cacatgcagc atgtatcaaa
     6421 attaatttgg ttttttttct taagtattta cattaaatgg ccatagttgc attaatgaat
     6481 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6541 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6601 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6661 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6721 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6781 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6841 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6901 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6961 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7021 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7081 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7141 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7201 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7261 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7321 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7381 gatcttcacc tagatccttt tgcggccggc cgcaaatcaa tctaaagtat atatgagtaa
     7441 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7501 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7561 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7621 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7681 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7741 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7801 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7861 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7921 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7981 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     8041 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     8101 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     8161 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     8221 tttactttca ccagcgtttc tgggtgagca aa
//
