LOCUS       sequence-418247-        8057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression in insect cells of TBK1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8057)
  AUTHORS   Adriaenssens E, Nguyen TN, Sawa-Makarska J, Khuu G, Schuschnig M, 
            Shoebridge S, Watts EM, Csalyi KD, Padman BS, Lazarou M, Martens S
  TITLE     Control of mitophagy initiation and progression by the TBK1 adaptors
            NAP1 and SINTBAD
  JOURNAL   bioRxiv September 25, 202
REFERENCE   2  (bases 1 to 8057)
  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: "bioRxiv 
            September 25, 202"
COMMENT     Sequence Label: pFastBac-Dual- GST-TEV-TBK1
FEATURES             Location/Qualifiers
     source          1..8057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..556
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(189..208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     398..419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        582..686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(687..755,756..1547)
                     /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     complement(905..924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1718..2306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2207..2226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2460..2477
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2611..2835
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2902..3435)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3624..3629,3630..3646,3647..3652))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    4160..4208
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4233..4252
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(4344..4453)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        4477..4568
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4493..4510
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4611..5267
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MASPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
                     KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIR
                     YGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVV
                     LYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     5225..5247
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             5274..5291
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5301..5321
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    7647..7781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7684..7703)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7738..7757
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7810..7975
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      361 ttttcgccct ttgacgttgg agtccacgtt cttaatagtg gactcttgtt ccaaactgga
      421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      541 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      601 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      661 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      721 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      781 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      841 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      901 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
      961 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1021 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1081 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1141 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1201 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1261 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1321 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1381 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1441 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2461 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     2521 tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct ggcaaaatcg gttacggttg
     2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
     2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
     2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
     2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
     2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
     2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
     3061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat
     3121 ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc caacaaccgc
     3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
     3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
     3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
     3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     3421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     3481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
     3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc
     3661 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg
     3721 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg
     3781 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg cccttgcttc
     3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
     3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
     3961 atagttctag tggttggcta cgtacccgta gtggctatgg cagggcttgc cgccccgacg
     4021 ttggctgcga gccctgggcc ttcacccgaa cttgggggtt ggggtgggga aaaggaagaa
     4081 acgcgggcgt attggtccca atggggtctc ggtggggtat cgacagagtg ccagccctgg
     4141 gaccgaaccc cgcgtttatg aacaaacgac ccaacacccg tgcgttttat tctgtctttt
     4201 tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct
     4261 cccccatctc ccggtaccgc atgctatgca tcagctgcta gcaccatggc tcgagatccc
     4321 gggtgatcaa gtcttcgtcg agtgattgta aataaaatgt aatttacagt atagtatttt
     4381 aattaatata caaatgattt gataataatt cttatttaac tataatatat tgtgttgggt
     4441 tgaattaaag gtccgtatac tccggaatat taatagatca tggagataat taaaatgata
     4501 accatctcgc aaataaataa gtattttact gttttcgtaa cagttttgta ataaaaaaac
     4561 ctataaatat tccggattat tcataccgtc ccaccatcgg gcgcggatcc atggcatccc
     4621 ctatactagg ttattggaaa attaagggcc ttgtgcaacc cactcgactt cttttggaat
     4681 atcttgaaga aaaatatgaa gagcatttgt atgagcgcga tgaaggtgat aaatggcgaa
     4741 acaaaaagtt tgaattgggt ttggagtttc ccaatcttcc ttattatatt gatggtgatg
     4801 ttaaattaac acagtctatg gccatcatac gttatatagc tgacaagcac aacatgttgg
     4861 gtggttgtcc aaaagagcgt gcagagattt caatgcttga aggagcggtt ttggatatta
     4921 gatacggtgt ttcgagaatt gcatatagta aagactttga aactctcaaa gttgattttc
     4981 ttagcaagct acctgaaatg ctgaaaatgt tcgaagatcg tttatgtcat aaaacatatt
     5041 taaatggtga tcatgtaacc catcctgact tcatgttgta tgacgctctt gatgttgttt
     5101 tatacatgga cccaatgtgc ctggatgcgt tcccaaaatt agtttgtttt aaaaaacgta
     5161 ttgaagctat cccacaaatt gataagtact tgaaatccag caagtatata gcatggcctt
     5221 tgcagggctg gcaagccacg tttggtggtg gcgaccatcc tccaaaatcg gatctggttc
     5281 cgcgtggatc aggatcagga gagaatttat actttcaggg agcgcgcatg caatccacat
     5341 ccaaccacct ctggctcctc tcggacattc tcggacaagg cgctaccgct aacgtgttca
     5401 ggggaaggca caagaagacc ggtgacctgt tcgccatcaa ggtgttcaac aacatctcat
     5461 tcctgcgccc tgtggacgtc cagatgcgtg agttcgaagt gctgaagaag ctgaaccaca
     5521 agaacatcgt caagctgttc gccatcgagg aagaaaccac tacccgccac aaggtgctga
     5581 tcatggagtt ctgcccctgc ggttccctgt acactgtcct ggaagagcct tccaacgctt
     5641 acggtctgcc cgagagcgag ttcctgatcg tcctgcgtga cgtggtcggt ggcatgaacc
     5701 acctgaggga aaacggcatc gtgcacagag acatcaagcc aggaaacatc atgcgcgtga
     5761 tcggcgagga cggacagagc gtctacaagc tgaccgactt cggcgctgcc agagagctgg
     5821 aagacgacga acagttcgtc tctctgtacg gaactgaaga gtacctgcac cctgacatgt
     5881 acgagagggc cgtgctgaga aaggaccacc agaagaagta cggtgctacc gtggacctgt
     5941 ggtctatcgg cgtcaccttc taccacgctg ccactggatc actgcctttc cgccccttcg
     6001 aaggtcctcg ccgtaacaag gaggtcatgt acaagatcat cactggcaag ccctctggcg
     6061 ccatctcagg agtccagaag gctgaaaacg gtcccatcga ctggagcggc gacatgccag
     6121 tgagctgctc tctgtcccgc ggtctgcaag tgctgctgac cccagtcctg gccaacatcc
     6181 tggaagctga ccaggagaag tgctggggtt tcgaccagtt cttcgctgaa acctccgaca
     6241 tcctgcacag aatggtcatc cacgtcttca gcctgcaaca gatgactgcc cacaagattt
     6301 acatccactc ttacaacact gctactatct tccacgaact ggtctacaag cagaccaaga
     6361 tcatctccag caaccaggaa ctgatctacg agggaaggag actggtgctg gagcccggtc
     6421 gcctggctca gcacttccca aagactaccg aagagaaccc tatcttcgtg gtgtcccgcg
     6481 agccactgaa cactatcggc ctgatctacg agaagatcag cctgcccaag gtccacccac
     6541 gttacgacct ggacggcgac gctagcatgg ctaaggccat caccggagtg gtctgctacg
     6601 cttgccgcat cgcttcaact ctgctgctgt accaggaact gatgcgcaag ggaatccgtt
     6661 ggctgatcga gctgatcaag gacgactaca acgaaactgt gcacaagaag accgaggtgg
     6721 tcatcactct ggacttctgc atccgtaaca tcgaaaagac cgtgaaggtc tacgagaagc
     6781 tgatgaagat caacctggag gctgccgaac tgggagagat cagcgacatc cacactaagc
     6841 tgctgcgcct gtcttcatcc cagggcacta tcgaaacctc cctgcaagac atcgactccc
     6901 gtctgagccc aggaggttca ctggctgacg cttgggctca ccaggaaggc acccacccta
     6961 aggacaggaa cgtggagaag ctgcaagtcc tgctgaactg catgactgaa atctactacc
     7021 agttcaagaa ggacaaggcc gagcgccgtc tggcttacaa cgaagagcag atccacaagt
     7081 tcgacaagca gaagctgtac taccacgcca ccaaggctat gactcacttc accgacgaat
     7141 gcgtcaagaa gtacgaggct ttcctgaaca agagcgaaga gtggatcagg aagatgctgc
     7201 acctgagaaa gcagctgctg tccctgacca accagtgctt cgacatcgaa gaggaagtga
     7261 gcaagtacca ggagtacacc aacgagctgc aagagactct gcctcagaag atgttcactg
     7321 ccagctctgg tatcaagcac actatgaccc caatctaccc ttcatccaac accctggtgg
     7381 aaatgactct gggtatgaag aagctgaagg aggaaatgga gggcgtggtc aaggaactgg
     7441 ctgagaacaa ccacatcttg gaaagattcg gtagcctgac gatggatggt ggactgagaa
     7501 acgtggactg tctgtaaaag cttgtcgaga agtactagag gatcataatc agccatacca
     7561 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     7621 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     7681 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     7741 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgatcactg cttgagccta
     7801 ggagatccga accagataag tgaaatctag ttccaaacta ttttgtcatt tttaattttc
     7861 gtattagctt acgacgctac acccagttcc catctatttt gtcactcttc cctaaataat
     7921 ccttaaaaac tccatttcca cccctcccag ttcccaacta ttttgtccgc ccacagcggg
     7981 gcatttttct tcctgttatg tttttaatca aacatcctgc caactccatg tgacaaaccg
     8041 tcatcttcgg ctacttt
//
