LOCUS       sequence-233070-        8228 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of LIR/UBD double mutant  OPTN .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8228)
  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 8228)
  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-OPTN(F178A/D474N)
FEATURES             Location/Qualifiers
     source          1..8228
                     /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"
     CDS             3170..3181
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4780..5249
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5379..5396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5550..6138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5630..5649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6302..7093,7094..7162))
                     /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     6925..6944
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7163..7267)
                     /gene="bla"
                     /label=AmpR promoter
     mobile_element  7412..8179
                     /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 cttggtccca tcaacctctc agctgcctca
     2881 ctgaaaagga ggacagcccc agtgaaagca caggaaatgg acccccccac ctggcccacc
     2941 caaacctgga cacgtttacc ccggaggagc tgctgcagca gatgaaagag ctcctgaccg
     3001 agaaccacca gctgaaagaa gccatgaagc taaataatca agccatgaaa gggagatttg
     3061 aggagctttc ggcctggaca gagaaacaga aggaagaacg ccagtttttt gagatacaga
     3121 gcaaagaagc aaaagagcgt ctaatggcct tgagtcatga gaatgagaaa ttgaaggaag
     3181 agcttggaaa actaaaaggg aaatcagaaa ggtcatctga ggaccccact gatgactcca
     3241 ggcttcccag ggccgaagcg gagcaggaaa aggaccagct caggacccag gtggtgaggc
     3301 tacaagcaga gaaggcagac ctgttgggca tcgtgtctga actgcagctc aagctgaact
     3361 ccagcggctc ctcagaagat tccgctgttg aaattaggat ggctgaagga gaagcagaag
     3421 ggtcagtaaa agaaatcaag catagtcctg ggcccacgag aacagtctcc actggcacgg
     3481 cattgtctaa atataggagc agatctgcag atggggccaa gaattacttc gaacatgagg
     3541 agttaactgt gagccagctc ctgctgtgcc taagggaagg gaatcagaag gtggagagac
     3601 ttgaagttgc actcaaggag gccaaagaaa gagtttcaga ttttgaaaag aaaacaagta
     3661 atcgttctga gattgaaacc cagacagagg ggagcacaga gaaagagaat gatgaagaga
     3721 aaggcccgga gactgttgga agcgaagtgg aagcactgaa cctccaggtg acatctctgt
     3781 ttaaggagct tcaagaggct catacaaaac tcagcgaagc tgagctaatg aagaagagac
     3841 ttcaagaaaa gtgtcaggcc cttgaaagga aaaattctgc aattccatca gagttgaatg
     3901 aaaagcaaga gcttgtttat actaacaaaa agttagagct acaagtggaa agcatgctat
     3961 cagaaatcaa aatggaacag gctaaaacag aggatgaaaa gtccaaatta actgtgctac
     4021 agatgacaca caacaagctt cttcaagaac ataataatgc attgaaaaca attgaggaac
     4081 taacaagaaa agagtcagaa aaagtggaca gggcagtgct gaaggaactg agtgaaaaac
     4141 tggaactggc agagaaggct ctggcttcca aacagctgca aatggatgaa atgaagcaaa
     4201 ccattgccaa gcaggaagag gacctggaaa ccatgaccat cctcagggct cagatggaag
     4261 tttactgttc taattttcat gctgaaagag cagcgagaga gaaaattcat gaggaaaagg
     4321 agcaactggc attgcagctg gcagttctgc tgaaagagaa tgatgctttc gaagacggag
     4381 gcaggcagtc cttgatggag atgcagagtc gtcatggggc gagaacaagt gactctgacc
     4441 agcaggctta ccttgttcaa agaggagctg aggacaggga ctggcggcaa cagcggaata
     4501 ttccgattca ttcctgcccc aagtgtggag aggttctgcc tgacatagac acgttacaga
     4561 ttcacgtgat ggattgcatc atttaaggat ccaccggatc tagataactg atcacgataa
     4621 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
     4681 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa
     4741 tagagaagtt cagatcaagg tcaggaacag atggaacagc tgaatatggg ccaaacagga
     4801 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatgga acagctgaat
     4861 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     4921 atggtcccca gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag
     4981 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     5041 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     5101 ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc
     5161 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
     5221 attgactacc cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt aatttggttt
     5281 tttttcttaa gtatttacat taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg
     5341 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5401 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5461 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5521 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5581 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5641 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5701 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5761 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5821 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5881 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5941 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6001 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6061 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6121 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6181 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6241 atccttttgc ggccggccgc aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6301 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6361 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6421 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6481 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6541 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6601 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6661 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6721 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6781 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6841 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6901 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6961 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7021 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7081 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7141 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7201 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7261 ttccgcgcac atttccccga aaagtgccac cagctttgct cttaggagtt tcctaataca
     7321 tcccaaactc aaatatataa agcatttgac ttgttctatg ccctaggggg cggggggaag
     7381 ctaagccagc tttttttaac atttaaaatg tggtaatgac tccaacttat tgatagtgtt
     7441 ttatgttcag ataatgcccg atgactttgt catgcagctc caccgatttt gagaacgaca
     7501 gcgacttccg tcccagccgt gccaggtgct gcctcagatt caggttatgc cgctcaattc
     7561 gctgcgtata tcgcttgctg attacgtgca gctttccctt caggcgggat tcatacagcg
     7621 gccagccatc cgtcatccat atcaccacgt caaagggtga cagcaggctc ataagacgcc
     7681 ccagcgtcgc catagtgcgt tcaccgaata cgtgcgcaac aaccgtcttc cggagactgt
     7741 catacgcgta aaacagccag cgctggcgcg atttagcccc gacatagccc cactgttcgt
     7801 ccatttccgc gcagacgatg acgtcactgc ccggctgtat gcgcgaggtt accgactgcg
     7861 gcctgagttt tttaagtgac gtaaaatcgt gttgaggcca acgcccataa tgcgggctgt
     7921 tgcccggcat ccaacgccat tcatggccat atcaatgatt ttctggtgcg taccgggttg
     7981 agaagcggtg taagtgaact gcagttgcca tgttttacgg cagtgagagc agagatagcg
     8041 ctgatgtccg gcggtgcttt tgccgttacg caccaccccg tcagtagctg aacaggaggg
     8101 acagctgata gaaacagaag ccactggagc acctcaaaaa caccatcata cactaaatca
     8161 gtaagttggc agcatcacct taaaatgtta attccatttt aaatgcacag atgtttttat
     8221 ttcataag
//
