LOCUS       sequence-180062-        6989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express TBK1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6989)
  AUTHORS   Marazioti A, Lilis I, Vreka M, Apostolopoulou H, Kalogeropoulou A, 
            Giopanou I, Giotopoulou GA, Krontira AC, Iliopoulou M, Kanellakis 
            NI, Agalioti T, Giannou AD, Jones-Paris C, Iwakura Y, Kardamakis D, 
            Blackwell TS, Taraviras S, Spella M, Stathopoulos GT
  TITLE     Myeloid-derived interleukin-1beta drives oncogenic KRAS-NF-kappaBeta
            addiction in malignant pleural effusion.
  JOURNAL   Nat Commun. 2018 Feb 14;9(1):672. doi: 10.1038/s41467-018-03051-z.
  PUBMED    29445180
REFERENCE   2  (bases 1 to 6989)
  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: "2018-02-14"; volume: "9(1):672. doi"; pages: " 
            10.1038/s41467-018-03051-z"
COMMENT     Sequence Label: pTBK1
FEATURES             Location/Qualifiers
     source          1..6989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..592
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     493..512
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        767..1070
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1071..1274
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1225..1245
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1313..1322
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1319..1321,1322..1324,1325..2035)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1319..1321,1322..1324,1325..2035)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1364..1385)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1625..1644)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1972..1993
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      4316..4325
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4320..4329
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4483..4604
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4520..4539)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4574..4593
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4611..5066)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4748..4769)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4960..4979
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5093..5197
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5199..5556
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5199..5390
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5222..5242)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5407..5542
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5591..6385
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5591..6385
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5645..5664)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6255..6274
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6573..6592)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6617..6664
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
       61 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
      121 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
      181 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
      241 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
      301 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
      361 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
      421 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      481 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      541 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      601 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      661 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
      721 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      781 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      841 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      901 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      961 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     1021 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     1081 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     1141 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     1201 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     1261 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat
     1321 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1381 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1441 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1501 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     1561 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1621 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1681 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1741 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1801 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1861 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1921 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1981 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg
     2041 actcagatct cgagctcaag cttcgaattc tgcagtcgac ctgcaggcgg ccgcgaattc
     2101 actagtgatt atgcagagca cctccaacca tctgtggctc ctgtctgata tcctaggcca
     2161 gggggccact gcaaatgtct tccgaggaag gcataagaaa actggtgatc tctatgctgt
     2221 caaagtattt aataacataa gcttccttcg cccagtggat gttcaaatga gagaatttga
     2281 agtgttaaaa aaactcaatc acaaaaacat tgtcaagtta tttgctattg aagaggagac
     2341 aacaacaaga cataaagtgc ttattatgga gttttgtccc tgtgggagtt tatacactgt
     2401 tctagaggag ccgtccaatg cgtatggact tccagaatca gaatttctca ttgtcttacg
     2461 agatgtggtg ggcgggatga atcatctccg agagaacggc atagtgcacc gagatatcaa
     2521 gccaggcaac atcatgcgcg tcatagggga ggacggccag tctgtgtaca aactcacgga
     2581 tttcggcgcc gctcgagagc tggaggacga tgagcagttt gtgtctctgt acggcacaga
     2641 agagtacctg catccggaca tgtatgaaag ggcagtgcta agaaaggacc atcagaagaa
     2701 gtacggggct accgttgatc tgtggagtgt tggagtgaca ttctaccatg cagccacggg
     2761 gtcgctgccg tttagaccct tcgaggggcc acggaggaac aaagaagtaa tgtataaaat
     2821 aatcactggg aagccgtctg gtgcaatatc tggagtacag aaagcagaaa acggaccaat
     2881 tgactggagt ggagacatgc ctctctcctg tagtctttct cagggtcttc aggcactgct
     2941 taccccagtt cttgcaaaca tacttgaagc tgatcaggag aagtgctggg gttttgacca
     3001 gttctttgca gagaccagtg atgtgcttca ccgaatggta atccatgtct tctcgctaca
     3061 acacatgacg gcgcataaga tttacattca cagctataac actgctgctg tgttccatga
     3121 actggtctat aaacaaacca agattgtttc ctcaaatcaa gaacttatct acgaaggacg
     3181 acgcttagtc ctagaactcg gacgactagc ccagcatttt cctaaaacca cagaggaaaa
     3241 tcctatcttt gtcacgagcc gggaacaact caataccgta ggactgagat atgaaaaaat
     3301 ttccctccct aaaatacatc cacgctatga tctggatggg gacgccagca tggccaaggc
     3361 agtgacgggg gttgtgtgct acgcctgcag aactgccagt accctgctgc tctatcaaga
     3421 attaatgcga aagggggtac ggtggctggt tgaactggtt aaggatgatt acaacgagac
     3481 cgtccacaag aagacggagg tagtgatcac actggatttc tgcatcagga acattgagaa
     3541 gactgtgaaa gtgtatgaga agttgatgaa ggtcaacctg gaagccgcag agctgggtga
     3601 gatttcagac atacacacca agctgctgag actttccagt tctcagggaa caatagaaag
     3661 cagtcttcag gacatcagca gcaggctgtc tccagggggc ttgctggccg acacctgggc
     3721 acatcaagaa ggcacgcatc caagagacag gaatgtagaa aaactgcagg tcctgttgaa
     3781 ctgcatcaca gagatttact atcagttcaa aaaagacaaa gcagaacgca gactagctta
     3841 taatgaagaa cagatccaca aatttgataa gcaaaaattg tattaccatg ccacaaaagc
     3901 aatgagccac ttctcagaag aatgtgttag aaagtatgaa gcgtttaaag ataagtcgga
     3961 agagtggatg agaaagatgc ttcatcttag gaagcagctg ttatcgctaa ctaatcagtg
     4021 tttcgatatc gaagaggaag tgtccaagta tcaagactat actaacgagt tacaagaaac
     4081 tctgcctcag aaaatgctcg cagcctccgg cggcgtcaag cacgccatgg ccccgatcta
     4141 ccccagctct aacaccttag tggagatgac tcttggtatg aagaagttaa aggaggagat
     4201 ggaaggcgtg gttaaggagc tggccgagaa caatcatatt ttagaaaggt ttgggtcttt
     4261 aacaatggat ggtggccttc gcaatgtgga ctgtctttag aatcgaattc ccgcggccgc
     4321 catggcggcc gggagcatgc gacgtcgggc ccgggatcca ccggatctag ataactgatc
     4381 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     4441 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     4501 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4561 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag
     4621 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     4681 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     4741 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     4801 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     4861 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     4921 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     4981 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     5041 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     5101 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5161 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     5221 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5281 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5341 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5401 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5461 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5521 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     5581 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5641 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5701 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5761 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5821 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5881 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5941 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     6001 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     6061 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     6121 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     6181 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     6241 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     6301 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     6361 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     6421 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6481 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6541 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     6601 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     6661 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     6721 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     6781 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     6841 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6901 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6961 tcagaccccg tagaaaagat caaaggatc
//
