LOCUS       sequence-232992-        8225 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of UBD mutant OPTN .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8225)
  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 8225)
  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(D474N)
FEATURES             Location/Qualifiers
     source          1..8225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             348..817
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    882..1239
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1304..1720
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1650..1672
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1688..1704
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1730..2104
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     2042..2058
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      2107..2116
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2113..2820
                     /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(2258..2276)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2526..2545
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3162..3173
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4772..5241
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5371..5388)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5542..6130)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5622..5641)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6294..7085,7086..7154))
                     /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     6917..6936
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7155..7259)
                     /gene="bla"
                     /label=AmpR promoter
     mobile_element  7404..8171
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
ORIGIN
        1 caatgtgcat gaatgctgca atattcctgt taccaaagct agtataaata aaaatagata
       61 aacgtggaaa ttacttagag tttctgtcat taacgtttcc ttcctcagtt gacaacataa
      121 atgcgctgct gagaagccag tttgcatctg tcaggatcaa tttcccatta tgccagtcat
      181 attaattact agtcaattag ttgattttta tttttgacat atacatgtga aagaccccac
      241 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
      301 actgagaata gaaaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
      361 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
      421 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
      481 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
      541 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
      601 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
      661 acccctcact cggcgcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
      721 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
      781 tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg
      841 gagacccctg cccagggacc accgacccac caccgggagg taagctggcc agcaacttat
      901 ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg tcggtactag
      961 ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt cggaacaccc
     1021 ggccgcaacc ctgggagacg tcccagggac ttcgggggcc gtttttgtgg cccgacctga
     1081 gtccaaaaat cccgatcgtt ttggactctt tggtgcaccc cccttagagg agggatatgt
     1141 ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc
     1201 ggtttgggac cgaagccgcg ccgcgcgtct tgtctgctgc agcatcgttc tgtgttgtct
     1261 ctgtctgact gtgtttctgt atttgtctga aaatatgggc ccgggccaga ctgttaccac
     1321 tcccttaagt ttgaccttag gtcactggaa agatgtcgag cggatcgctc acaaccagtc
     1381 ggtagatgtc aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa
     1441 cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa
     1501 ggtcttttca cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga
     1561 agccttggct tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc
     1621 tcctcttcct ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg
     1681 atcctccctt tatccagccc tcactccttc tctaggcgcc cccatatggc catatgagat
     1741 cttatatggg gcacccccgc cccttgtaaa cttccctgac cctgacatga caagagttac
     1801 taacagcccc tctctccaag ctcacttaca ggctctctac ttagtccagc acgaagtctg
     1861 gagacctctg gcggcagcct accaagaaca actggaccga ccggtggtac ctcaccctta
     1921 ccgagtcggc gacacagtgt gggtccgccg acaccagact aagaacctag aacctcgctg
     1981 gaaaggacct tacacagtcc tgctgaccac ccccaccgcc ctcaaagtag acggcatcgc
     2041 agcttggata cacgccgccc acgtgaaggc tgccgacccc gggggtggac catcctctag
     2101 actgccgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     2161 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     2221 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     2281 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     2341 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     2401 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     2461 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     2521 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     2581 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     2641 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     2701 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     2761 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     2821 tccggactca gatctcgagc tcaagcttcc catcaacctc tcagctgcct cactgaaaag
     2881 gaggacagcc ccagtgaaag cacaggaaat ggaccccccc acctggccca cccaaacctg
     2941 gacacgttta ccccggagga gctgctgcag cagatgaaag agctcctgac cgagaaccac
     3001 cagctgaaag aagccatgaa gctaaataat caagccatga aagggagatt tgaggagctt
     3061 tcggcctgga cagagaaaca gaaggaagaa cgccagtttt ttgagataca gagcaaagaa
     3121 gcaaaagagc gtctaatggc cttgagtcat gagaatgaga aattgaagga agagcttgga
     3181 aaactaaaag ggaaatcaga aaggtcatct gaggacccca ctgatgactc caggcttccc
     3241 agggccgaag cggagcagga aaaggaccag ctcaggaccc aggtggtgag gctacaagca
     3301 gagaaggcag acctgttggg catcgtgtct gaactgcagc tcaagctgaa ctccagcggc
     3361 tcctcagaag attcctttgt tgaaattagg atggctgaag gagaagcaga agggtcagta
     3421 aaagaaatca agcatagtcc tgggcccacg agaacagtct ccactggcac ggcattgtct
     3481 aaatatagga gcagatctgc agatggggcc aagaattact tcgaacatga ggagttaact
     3541 gtgagccagc tcctgctgtg cctaagggaa gggaatcaga aggtggagag acttgaagtt
     3601 gcactcaagg aggccaaaga aagagtttca gattttgaaa agaaaacaag taatcgttct
     3661 gagattgaaa cccagacaga ggggagcaca gagaaagaga atgatgaaga gaaaggcccg
     3721 gagactgttg gaagcgaagt ggaagcactg aacctccagg tgacatctct gtttaaggag
     3781 cttcaagagg ctcatacaaa actcagcgaa gctgagctaa tgaagaagag acttcaagaa
     3841 aagtgtcagg cccttgaaag gaaaaattct gcaattccat cagagttgaa tgaaaagcaa
     3901 gagcttgttt atactaacaa aaagttagag ctacaagtgg aaagcatgct atcagaaatc
     3961 aaaatggaac aggctaaaac agaggatgaa aagtccaaat taactgtgct acagatgaca
     4021 cacaacaagc ttcttcaaga acataataat gcattgaaaa caattgagga actaacaaga
     4081 aaagagtcag aaaaagtgga cagggcagtg ctgaaggaac tgagtgaaaa actggaactg
     4141 gcagagaagg ctctggcttc caaacagctg caaatggatg aaatgaagca aaccattgcc
     4201 aagcaggaag aggacctgga aaccatgacc atcctcaggg ctcagatgga agtttactgt
     4261 tctaattttc atgctgaaag agcagcgaga gagaaaattc atgaggaaaa ggagcaactg
     4321 gcattgcagc tggcagttct gctgaaagag aatgatgctt tcgaagacgg aggcaggcag
     4381 tccttgatgg agatgcagag tcgtcatggg gcgagaacaa gtgactctga ccagcaggct
     4441 taccttgttc aaagaggagc tgaggacagg gactggcggc aacagcggaa tattccgatt
     4501 cattcctgcc ccaagtgtgg agaggttctg cctgacatag acacgttaca gattcacgtg
     4561 atggattgca tcatttaagg atccaccgga tctagataac tgatcacgat aaaataaaag
     4621 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
     4681 ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag
     4741 ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     4801 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca
     4861 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     4921 cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     4981 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     5041 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc
     5101 gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     5161 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     5221 cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa ttaatttggt tttttttctt
     5281 aagtatttac attaaatggc catagttgca ttaatgaatc ggccaacgcg cggggagagg
     5341 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5401 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5461 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5521 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5581 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5641 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5701 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5761 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5821 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5881 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5941 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6001 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6061 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6121 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6181 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6241 gcggccggcc gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6301 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6361 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6421 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6481 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6541 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6601 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6661 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6721 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6781 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6841 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6901 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6961 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7021 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7081 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7141 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7201 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7261 acatttcccc gaaaagtgcc accagctttg ctcttaggag tttcctaata catcccaaac
     7321 tcaaatatat aaagcatttg acttgttcta tgccctaggg ggcgggggga agctaagcca
     7381 gcttttttta acatttaaaa tgtggtaatg actccaactt attgatagtg ttttatgttc
     7441 agataatgcc cgatgacttt gtcatgcagc tccaccgatt ttgagaacga cagcgacttc
     7501 cgtcccagcc gtgccaggtg ctgcctcaga ttcaggttat gccgctcaat tcgctgcgta
     7561 tatcgcttgc tgattacgtg cagctttccc ttcaggcggg attcatacag cggccagcca
     7621 tccgtcatcc atatcaccac gtcaaagggt gacagcaggc tcataagacg ccccagcgtc
     7681 gccatagtgc gttcaccgaa tacgtgcgca acaaccgtct tccggagact gtcatacgcg
     7741 taaaacagcc agcgctggcg cgatttagcc ccgacatagc cccactgttc gtccatttcc
     7801 gcgcagacga tgacgtcact gcccggctgt atgcgcgagg ttaccgactg cggcctgagt
     7861 tttttaagtg acgtaaaatc gtgttgaggc caacgcccat aatgcgggct gttgcccggc
     7921 atccaacgcc attcatggcc atatcaatga ttttctggtg cgtaccgggt tgagaagcgg
     7981 tgtaagtgaa ctgcagttgc catgttttac ggcagtgaga gcagagatag cgctgatgtc
     8041 cggcggtgct tttgccgtta cgcaccaccc cgtcagtagc tgaacaggag ggacagctga
     8101 tagaaacaga agccactgga gcacctcaaa aacaccatca tacactaaat cagtaagttg
     8161 gcagcatcac cttaaaatgt taattccatt ttaaatgcac agatgttttt atttcataag
     8221 ggttt
//
