LOCUS       sequence-232961-        8450 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of iRFP-tagged LIR/UBD double mutant OPTN .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8450)
  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 8450)
  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-OPTN(F178A/D474N)
FEATURES             Location/Qualifiers
     source          1..8450
                     /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"
     CDS             3392..3403
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5002..5471
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5601..5618)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5772..6360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5852..5871)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6524..7315,7316..7384))
                     /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     7147..7166
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7385..7489)
                     /gene="bla"
                     /label=AmpR promoter
     mobile_element  7634..8401
                     /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 tcaagcttcc catcaacctc tcagctgcct cactgaaaag gaggacagcc
     3121 ccagtgaaag cacaggaaat ggaccccccc acctggccca cccaaacctg gacacgttta
     3181 ccccggagga gctgctgcag cagatgaaag agctcctgac cgagaaccac cagctgaaag
     3241 aagccatgaa gctaaataat caagccatga aagggagatt tgaggagctt tcggcctgga
     3301 cagagaaaca gaaggaagaa cgccagtttt ttgagataca gagcaaagaa gcaaaagagc
     3361 gtctaatggc cttgagtcat gagaatgaga aattgaagga agagcttgga aaactaaaag
     3421 ggaaatcaga aaggtcatct gaggacccca ctgatgactc caggcttccc agggccgaag
     3481 cggagcagga aaaggaccag ctcaggaccc aggtggtgag gctacaagca gagaaggcag
     3541 acctgttggg catcgtgtct gaactgcagc tcaagctgaa ctccagcggc tcctcagaag
     3601 attccgctgt tgaaattagg atggctgaag gagaagcaga agggtcagta aaagaaatca
     3661 agcatagtcc tgggcccacg agaacagtct ccactggcac ggcattgtct aaatatagga
     3721 gcagatctgc agatggggcc aagaattact tcgaacatga ggagttaact gtgagccagc
     3781 tcctgctgtg cctaagggaa gggaatcaga aggtggagag acttgaagtt gcactcaagg
     3841 aggccaaaga aagagtttca gattttgaaa agaaaacaag taatcgttct gagattgaaa
     3901 cccagacaga ggggagcaca gagaaagaga atgatgaaga gaaaggcccg gagactgttg
     3961 gaagcgaagt ggaagcactg aacctccagg tgacatctct gtttaaggag cttcaagagg
     4021 ctcatacaaa actcagcgaa gctgagctaa tgaagaagag acttcaagaa aagtgtcagg
     4081 cccttgaaag gaaaaattct gcaattccat cagagttgaa tgaaaagcaa gagcttgttt
     4141 atactaacaa aaagttagag ctacaagtgg aaagcatgct atcagaaatc aaaatggaac
     4201 aggctaaaac agaggatgaa aagtccaaat taactgtgct acagatgaca cacaacaagc
     4261 ttcttcaaga acataataat gcattgaaaa caattgagga actaacaaga aaagagtcag
     4321 aaaaagtgga cagggcagtg ctgaaggaac tgagtgaaaa actggaactg gcagagaagg
     4381 ctctggcttc caaacagctg caaatggatg aaatgaagca aaccattgcc aagcaggaag
     4441 aggacctgga aaccatgacc atcctcaggg ctcagatgga agtttactgt tctaattttc
     4501 atgctgaaag agcagcgaga gagaaaattc atgaggaaaa ggagcaactg gcattgcagc
     4561 tggcagttct gctgaaagag aatgatgctt tcgaagacgg aggcaggcag tccttgatgg
     4621 agatgcagag tcgtcatggg gcgagaacaa gtgactctga ccagcaggct taccttgttc
     4681 aaagaggagc tgaggacagg gactggcggc aacagcggaa tattccgatt cattcctgcc
     4741 ccaagtgtgg agaggttctg cctgacatag acacgttaca gattcacgtg atggattgca
     4801 tcatttaagg atccaccgga tctagataac tgatcacgat aaaataaaag attttattta
     4861 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     4921 taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa
     4981 ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
     5041 cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat
     5101 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt
     5161 ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
     5221 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     5281 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc
     5341 cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
     5401 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
     5461 ggggtctttc acatgcagca tgtatcaaaa ttaatttggt tttttttctt aagtatttac
     5521 attaaatggc catagttgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5581 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5641 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5701 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5761 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5821 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5881 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5941 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6001 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6061 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6121 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6181 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6241 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6301 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6361 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6421 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt gcggccggcc
     6481 gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6541 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6601 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6661 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6721 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6781 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6841 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6901 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6961 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7021 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7081 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7141 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7201 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7261 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7321 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7381 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7441 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7501 gaaaagtgcc accagctttg ctcttaggag tttcctaata catcccaaac tcaaatatat
     7561 aaagcatttg acttgttcta tgccctaggg ggcgggggga agctaagcca gcttttttta
     7621 acatttaaaa tgtggtaatg actccaactt attgatagtg ttttatgttc agataatgcc
     7681 cgatgacttt gtcatgcagc tccaccgatt ttgagaacga cagcgacttc cgtcccagcc
     7741 gtgccaggtg ctgcctcaga ttcaggttat gccgctcaat tcgctgcgta tatcgcttgc
     7801 tgattacgtg cagctttccc ttcaggcggg attcatacag cggccagcca tccgtcatcc
     7861 atatcaccac gtcaaagggt gacagcaggc tcataagacg ccccagcgtc gccatagtgc
     7921 gttcaccgaa tacgtgcgca acaaccgtct tccggagact gtcatacgcg taaaacagcc
     7981 agcgctggcg cgatttagcc ccgacatagc cccactgttc gtccatttcc gcgcagacga
     8041 tgacgtcact gcccggctgt atgcgcgagg ttaccgactg cggcctgagt tttttaagtg
     8101 acgtaaaatc gtgttgaggc caacgcccat aatgcgggct gttgcccggc atccaacgcc
     8161 attcatggcc atatcaatga ttttctggtg cgtaccgggt tgagaagcgg tgtaagtgaa
     8221 ctgcagttgc catgttttac ggcagtgaga gcagagatag cgctgatgtc cggcggtgct
     8281 tttgccgtta cgcaccaccc cgtcagtagc tgaacaggag ggacagctga tagaaacaga
     8341 agccactgga gcacctcaaa aacaccatca tacactaaat cagtaagttg gcagcatcac
     8401 cttaaaatgt taattccatt ttaaatgcac agatgttttt atttcataag
//
