LOCUS       sequence-393048-        8006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for the expression of TBK1 carrying a kinase dead mutation 
            D135N (SMC Internal No. 1997).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8006)
  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 8006)
  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_HA-TBK1 (kinase dead - D135N)
FEATURES             Location/Qualifiers
     source          1..8006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             166..635
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    700..1057
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1122..1538
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1468..1490
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1506..1522
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1548..1922
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1860..1876
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             1934..1960
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             4371..4840
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4970..4987)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5141..5729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5221..5240)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5893..6684,6685..6753))
                     /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     6516..6535
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6754..6858)
                     /gene="bla"
                     /label=AmpR promoter
     mobile_element  7003..7770
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
ORIGIN
        1 taattactag tcaattagtt gatttttatt tttgacatat acatgtgaaa gaccccacct
       61 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac
      121 tgagaataga aaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa
      181 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
      241 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      301 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
      361 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      421 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      481 ccctcactcg gcgcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
      541 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
      601 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
      661 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggccag caacttatct
      721 gtgtctgtcc gattgtctag tgtctatgac tgattttatg cgcctgcgtc ggtactagtt
      781 agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttcg gaacacccgg
      841 ccgcaaccct gggagacgtc ccagggactt cgggggccgt ttttgtggcc cgacctgagt
      901 ccaaaaatcc cgatcgtttt ggactctttg gtgcaccccc cttagaggag ggatatgtgg
      961 ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg
     1021 tttgggaccg aagccgcgcc gcgcgtcttg tctgctgcag catcgttctg tgttgtctct
     1081 gtctgactgt gtttctgtat ttgtctgaaa atatgggccc gggccagact gttaccactc
     1141 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1201 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1261 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1321 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1381 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1441 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1501 cctcccttta tccagccctc actccttctc taggcgcccc catatggcca tatgagatct
     1561 tatatggggc acccccgccc cttgtaaact tccctgaccc tgacatgaca agagttacta
     1621 acagcccctc tctccaagct cacttacagg ctctctactt agtccagcac gaagtctgga
     1681 gacctctggc ggcagcctac caagaacaac tggaccgacc ggtggtacct cacccttacc
     1741 gagtcggcga cacagtgtgg gtccgccgac accagactaa gaacctagaa cctcgctgga
     1801 aaggacctta cacagtcctg ctgaccaccc ccaccgccct caaagtagac ggcatcgcag
     1861 cttggataca cgccgcccac gtgaaggctg ccgaccccgg gggtggacca tcctctagac
     1921 tgccgccacc atgtacccat acgatgttcc agattacgct tccggactca gatctcgagc
     1981 tcaagcttct cagagcactt ctaatcatct gtggctttta tctgatattt taggccaagg
     2041 agctactgca aatgtctttc gtggaagaca taagaaaact ggtgatttat ttgctatcaa
     2101 agtatttaat aacataagct tccttcgtcc agtggatgtt caaatgagag aatttgaagt
     2161 gttgaaaaaa ctcaatcaca aaaatattgt caaattattt gctattgaag aggagacaac
     2221 aacaagacat aaagtactta ttatggaatt ttgtccatgt gggagtttat acactgtttt
     2281 agaagaacct tctaatgcct atggactacc agaatctgaa ttcttaattg ttttgcgaga
     2341 tgtggtgggt ggaatgaatc atctacgaga gaatggtata gtgcaccgta atatcaagcc
     2401 aggaaatatc atgcgtgtta taggggaaga tggacagtct gtgtacaaac tcacagattt
     2461 tggtgcagct agagaattag aagatgatga gcagtttgtt tctctgtatg gcacagaaga
     2521 atatttgcac cctgatatgt atgagagagc agtgctaaga aaagatcatc agaagaaata
     2581 tggagcaaca gttgatcttt ggagcattgg ggtaacattt taccatgcag ctactggatc
     2641 actgccattt agaccctttg aagggcctcg taggaataaa gaagtgatgt ataaaataat
     2701 tacaggaaag ccttctggtg caatatctgg agtacagaaa gcagaaaatg gaccaattga
     2761 ctggagtgga gacatgcctg tttcttgcag tctttctcgg ggtcttcagg ttctacttac
     2821 ccctgttctt gcaaacatcc ttgaagcaga tcaggaaaag tgttggggtt ttgaccagtt
     2881 ttttgcagaa actagtgata tacttcaccg aatggtaatt catgtttttt cgctacaaca
     2941 aatgacagct cataagattt atatacatag ctataatact gctactatat ttcatgaact
     3001 ggtatataaa caaaccaaaa ttatttcttc aaatcaagaa cttatctacg aagggcgacg
     3061 cttagtctta gaacctggaa ggctggcaca acatttccct aaaactactg aggaaaaccc
     3121 tatatttgta gtaagccggg aacctctgaa taccatagga ttaatatatg aaaaaatttc
     3181 cctccctaaa gtacatccac gttatgattt agacggggat gctagcatgg ctaaggcaat
     3241 aacaggggtt gtgtgttatg cctgcagaat tgccagtacc ttactgcttt atcaggaatt
     3301 aatgcgaaag gggatacgat ggctgattga attaattaaa gatgattaca atgaaactgt
     3361 tcacaaaaag acagaagttg tgatcacatt ggatttctgt atcagaaaca ttgaaaaaac
     3421 tgtgaaagta tatgaaaagt tgatgaagat caacctggaa gcggcagagt taggtgaaat
     3481 ttcagacata cacaccaaat tgttgagact ttccagttct cagggaacaa tagaaaccag
     3541 tcttcaggat atcgacagca gattatctcc aggtggatca ctggcagacg catgggcaca
     3601 tcaagaaggc actcatccga aagacagaaa tgtagaaaaa ctacaagtcc tgttaaattg
     3661 catgacagag atttactatc agttcaaaaa agacaaagca gaacgtagat tagcttataa
     3721 tgaagaacaa atccacaaat ttgataagca aaaactgtat taccatgcca caaaagctat
     3781 gacgcacttt acagatgaat gtgttaaaaa gtatgaggca tttttgaata agtcagaaga
     3841 atggataaga aagatgcttc atcttaggaa acagttatta tcgctgacta atcagtgttt
     3901 tgatattgaa gaagaagtat caaaatatca agaatatact aatgagttac aagaaactct
     3961 gcctcagaaa atgtttacag cttccagtgg aatcaaacat accatgaccc caatttatcc
     4021 aagttctaac acattagtag aaatgactct tggtatgaag aaattaaagg aagagatgga
     4081 aggggtggtt aaagaacttg ctgaaaataa ccacatttta gaaaggtttg gctctttaac
     4141 catggatggt ggccttcgca acgttgactg tctttaggga tccaccggat ctagataact
     4201 gatcacgata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
     4261 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac
     4321 ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg
     4381 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     4441 aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     4501 gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca
     4561 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa
     4621 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
     4681 acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta
     4741 tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc
     4801 tcctctgagt gattgactac ccgtcagcgg gggtctttca catgcagcat gtatcaaaat
     4861 taatttggtt ttttttctta agtatttaca ttaaatggcc atagttgcat taatgaatcg
     4921 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4981 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5041 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5101 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5161 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5221 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5281 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5341 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5401 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5461 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5521 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5581 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5641 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5701 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5761 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5821 tcttcaccta gatccttttg cggccggccg caaatcaatc taaagtatat atgagtaaac
     5881 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5941 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6001 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6061 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6121 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6181 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6241 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6301 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6361 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6421 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6481 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6541 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6601 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6661 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6721 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6781 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6841 acaaataggg gttccgcgca catttccccg aaaagtgcca ccagctttgc tcttaggagt
     6901 ttcctaatac atcccaaact caaatatata aagcatttga cttgttctat gccctagggg
     6961 gcggggggaa gctaagccag ctttttttaa catttaaaat gtggtaatga ctccaactta
     7021 ttgatagtgt tttatgttca gataatgccc gatgactttg tcatgcagct ccaccgattt
     7081 tgagaacgac agcgacttcc gtcccagccg tgccaggtgc tgcctcagat tcaggttatg
     7141 ccgctcaatt cgctgcgtat atcgcttgct gattacgtgc agctttccct tcaggcggga
     7201 ttcatacagc ggccagccat ccgtcatcca tatcaccacg tcaaagggtg acagcaggct
     7261 cataagacgc cccagcgtcg ccatagtgcg ttcaccgaat acgtgcgcaa caaccgtctt
     7321 ccggagactg tcatacgcgt aaaacagcca gcgctggcgc gatttagccc cgacatagcc
     7381 ccactgttcg tccatttccg cgcagacgat gacgtcactg cccggctgta tgcgcgaggt
     7441 taccgactgc ggcctgagtt ttttaagtga cgtaaaatcg tgttgaggcc aacgcccata
     7501 atgcgggctg ttgcccggca tccaacgcca ttcatggcca tatcaatgat tttctggtgc
     7561 gtaccgggtt gagaagcggt gtaagtgaac tgcagttgcc atgttttacg gcagtgagag
     7621 cagagatagc gctgatgtcc ggcggtgctt ttgccgttac gcaccacccc gtcagtagct
     7681 gaacaggagg gacagctgat agaaacagaa gccactggag cacctcaaaa acaccatcat
     7741 acactaaatc agtaagttgg cagcatcacc ttaaaatgtt aattccattt taaatgcaca
     7801 gatgttttta tttcataagg gtttcaatgt gcatgaatgc tgcaatattc ctgttaccaa
     7861 agctagtata aataaaaata gataaacgtg gaaattactt agagtttctg tcattaacgt
     7921 ttccttcctc agttgacaac ataaatgcgc tgctgagaag ccagtttgca tctgtcagga
     7981 tcaatttccc attatgccag tcatat
//
