LOCUS       sequence-429271-        7169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of TBK1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7169)
  AUTHORS   Tudorica DA, Basak B, Puerta Cordova AS, Khuu G, Rose K, Lazarou M, 
            Holzbaur ELF, Hurley JH
  TITLE     A RAB7A phosphoswitch coordinates Rubicon Homology protein 
            regulation of Parkin-dependent mitophagy.
  JOURNAL   J Cell Biol. 2024 Jul 1;223(7):e202309015. doi: 
            10.1083/jcb.202309015. Epub 2024 May 10.
  PUBMED    38728007
REFERENCE   2  (bases 1 to 7169)
  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: "J Cell 
            Biol."; date: "2024-07-1"; volume: "223(7):e202309015. doi"; pages: 
            " 10.1083/jcb.202309015. Epub 2024 May 10"
COMMENT     Sequence Label: pCAG-TSF-TBK1
FEATURES             Location/Qualifiers
     source          1..7169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..663
                     /label=chicken beta-actin promoter
     intron          664..1680
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1604..1626
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1688..1707
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1755..1764
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1759..1768
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1770..1793
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1830..1853
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1860..1883
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4191..4210)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4256..4311
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4310..4329)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4672..4688)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4672..4688)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4685..4707)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4696..4712
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4720..4726,4727..4744,4745..4750))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4765..4786
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4845..5040
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4891..5026
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5046..5180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5083..5102)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5137..5156
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5248..5265)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5419..6007)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5499..5518)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6178..6969,6970..7038))
                     /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     6801..6820
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7039..7143)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg
      541 cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
      601 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
      661 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
      721 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      781 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
      841 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
      961 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     1021 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     1081 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
     1141 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
     1201 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg
     1261 gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc
     1321 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     1381 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     1441 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1501 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     1561 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     1621 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     1681 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcctcgat
     1741 cgacggtatc gatagccacc atggcggcct ggtcacaccc gcagttcgag aaaggtggag
     1801 gtgcccgagg tggatcggga ggtggatcgt ggagccaccc gcagttcgaa aaaggattcg
     1861 attacaagga tgacgatgac aagggtacca tgcagtctac ctccaaccat ctctggctcc
     1921 tcagtgatat tctcggtcag ggtgctacag ctaatgtctt tcggggacgc cataagaaga
     1981 ccggcgacct gttcgctatc aaggtgttca acaacatctc cttcctgcgc cccgtggacg
     2041 tgcagatgag ggagttcgaa gtgctgaaga agctgaacca caagaacatc gtgaagctgt
     2101 tcgctatcga ggaagaaacc accacccgcc acaaggtgct gatcatggag ttctgccctt
     2161 gcggttctct gtacaccgtg ctggaagagc cctccaacgc ctacggcctg cctgagtctg
     2221 aattcctgat cgtgctgagg gacgtggtgg gcggaatgaa ccacctgaga gaaaacggaa
     2281 tcgtgcaccg tgacatcaag cctggtaaca tcatgagagt gatcggagag gacggtcaga
     2341 gcgtgtacaa gctgaccgac ttcggagccg ctcgtgagct ggaagacgac gaacagttcg
     2401 tgagcctgta cggcaccgaa gagtacctgc accccgacat gtacgagaga gctgtgctgc
     2461 gtaaggacca ccagaagaag tatggtgcta ccgtggacct gtggagcatc ggagtgacct
     2521 tctaccacgc tgctaccggt tccctgccct tccgcccttt cgaaggtcca cgtaggaaca
     2581 aggaagtgat gtacaagatc atcaccggca agccttctgg agctatcagc ggtgtgcaga
     2641 aggccgagaa cggccctatc gactggagcg gagacatgcc agtgagctgc tccctgtcta
     2701 gaggtctgca ggtgctgctg acccctgtgc tggctaacat cctggaagcc gaccaggaga
     2761 agtgctgggg cttcgaccag ttcttcgccg aaaccagcga catcctgcac cgtatggtca
     2821 tccacgtgtt cagcctgcag cagatgaccg ctcacaagat ctacatccac tcctacaaca
     2881 ccgctaccat cttccacgag ctggtgtaca agcagaccaa gatcatcagc tccaaccagg
     2941 aactgatcta cgagggccgc cgcctggtgc tggagcctgg ccgcctggct cagcacttcc
     3001 caaagaccac cgaagagaac cccatcttcg tggtgtctag ggaaccactg aacaccatcg
     3061 gactgatcta cgagaagatc agcctgccta aggtgcaccc aagatacgac ctggacggag
     3121 acgcctccat ggccaaggct atcaccggtg tggtgtgcta cgcttgccgt atcgcctcta
     3181 ccctgctgct gtaccaggaa ctgatgcgca agggtatcag gtggctgatc gagctgatca
     3241 aggacgacta caacgaaacc gtgcacaaga agaccgaggt ggtcatcacc ctggacttct
     3301 gcatcaggaa catcgaaaag accgtgaagg tgtacgagaa gctgatgaag atcaacctgg
     3361 aggccgctga actgggcgag atcagcgaca tccacaccaa gctgctgcgc ctgtctagct
     3421 cccagggaac catcgaaacc agcctgcagg acatcgacag caggctgtcc ccaggtggct
     3481 ctctggctga cgcttgggct caccaggaag gaacccaccc caaggaccgc aacgtggaga
     3541 agctgcaggt gctgctgaac tgcatgaccg aaatctacta ccagttcaag aaggacaagg
     3601 ctgagcgcag gctggcctac aacgaagagc agatccacaa gttcgacaag cagaagctgt
     3661 actaccacgc taccaaggcc atgacccact tcaccgatga atgcgtgaag aagtacgagg
     3721 ccttcctgaa caagtccgaa gagtggatca gaaagatgct gcacctgcgt aagcagctgc
     3781 tgagcctgac caaccagtgc ttcgacatcg aagaggaagt gtccaagtac caggagtaca
     3841 ccaacgaact gcaggaaacc ctgccccaga agatgttcac cgcttctagc ggcatcaagc
     3901 acaccatgac cccaatctac ccctcctcta acaccctggt ggaaatgacc ctgggcatga
     3961 agaagctgaa ggaggaaatg gagggagtgg tgaaggaact ggccgagaac aaccacattc
     4021 tggaaagatt cggtagcctg actatggatg gcggtctgcg taatgtggac tgtctccccg
     4081 ggtgatatct taattaatta actcgaggaa ttcactcctc aggtgcaggc tgcctatcag
     4141 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
     4201 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     4261 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     4321 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     4381 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
     4441 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
     4501 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     4561 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     4621 cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct
     4681 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4741 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4801 actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca
     4861 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4921 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4981 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5041 aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5101 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5161 tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5221 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5281 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5341 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5401 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5461 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5521 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5581 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5641 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5701 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5761 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5821 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5881 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5941 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6001 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6061 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6121 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6181 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6241 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6301 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6361 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6421 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6481 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6541 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6601 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6661 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6721 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6781 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6841 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6901 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6961 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7021 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7081 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7141 gcgcacattt ccccgaaaag tgccacctg
//
