LOCUS       sequence-243303-        8434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of mouse Tbkbp1 (1-330).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8434)
  AUTHORS   Zhu L, Xie X, Zhang L, Wang H, Jie Z, Zhou X, Shi J, Zhao S, Zhang 
            B, Cheng X, Sun SC
  TITLE     TBK-binding protein 1 regulates IL-15-induced autophagy and NKT cell
            survival.
  JOURNAL   Nat Commun. 2018 Jul 18;9(1):2812. doi: 10.1038/s41467-018-05097-5.
  PUBMED    30022064
REFERENCE   2  (bases 1 to 8434)
  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: "2018-07-18"; volume: "9(1):2812. doi"; pages: " 
            10.1038/s41467-018-05097-5"
COMMENT     Sequence Label: pCLXSN(GFP)-HA-Tbkbp1 1-330
FEATURES             Location/Qualifiers
     source          1..8434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        74..453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        454..657
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     608..628
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             658..833
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    896..1095
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1296..1712
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1642..1664
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1680..1696
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1728..1754
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2769..2789)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2774..3103
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2954..3089
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(3162..3164,3165..3167,3168..3881)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3162..3164,3165..3167,3168..3878)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3207..3228)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3468..3487)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3815..3836
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3987..4580
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     intron          4925..4990
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5120..5140
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5412..5546
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5449..5468)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5503..5522
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6004..6138
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6203..6220)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6374..6962)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6454..6473)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7133..7924,7925..7993))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7756..7775
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7994..8098)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8166..8184
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     8344..8363
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
       61 acgccagcgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      121 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      181 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      301 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      361 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      421 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      481 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      541 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      601 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcaa
      661 taaaagagcc cacaacccct cactcggcgc gccagtcttc cgatagactg cgtcgcccgg
      721 gtacccgtat tcccaataaa gcctcttgct gtttgcatcc gaatcgtggt ctcgctgttc
      781 cttgggaggg tctcctctga gtgattgact acccacgacg ggggtctttc atttgggggc
      841 tcgtccggga tttggagacc cctgcccagg gaccaccgac ccaccaccgg gaggtaagct
      901 ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc tatgtttgat gttatgcgcc
      961 tgcgtctgta ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg
     1021 agttctgaac acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt
     1081 gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc
     1141 tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt
     1201 ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct
     1261 gtctgactgt gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa
     1321 gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg
     1381 tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat
     1441 ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt
     1501 cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg
     1561 cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc
     1621 ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc
     1681 tttatccagc cctcactcct tctctaggcg ccggaattat catggcctac ccttatgatg
     1741 tgccagatta tgcctctccc gaattcatgg agtccatgtt tgaagatgac atcagcatcc
     1801 tgacccaaga ggccttgggg cccagtgagg tgtggctgga tggtcctgga gacccgtccc
     1861 tgggagggga catgtgttct gcctcccact ttgccctcat cacagcctac ggagacatca
     1921 aggaaaggct aggaggcctg gagagggaga atgccaccct ccgccgtcgc ctcaaagtct
     1981 atgagatcaa gtatccgctg atcactgatt ttggagagga gcatggcttc cctctgtatg
     2041 aactcaagga tggctccctg ctggaggtgg agaaggtcag tctgcagcaa cgactcaacc
     2101 aattccaaca cgagttgcag aagagtaagg agcaggagga gcagctcggg gagatgatcc
     2161 aggcttacga gaaactgtgc gtggagaaaa gcgacttgga gacagagctg ggggagatgc
     2221 gggccctggt agagactcat ctacggcaga tctgtggtct ggagaaacag ctgcagcaac
     2281 agcagggcct ccgggacgca gctttctcca gcctgagccc cccagctgtc cccgcctcag
     2341 cctgtcctga cttggacctg cactatctgg ctctgagagg gggacctgcc ttgggtcatg
     2401 gttggcctgg ccctacgagt gtaagtgtga gcgagctgga gaggaggcgc ctggaagagg
     2461 ccctggaggc tgcgcagggg gaggctcggg gggctcagct ccgggaggag caactccagg
     2521 cggaatgcga gagacttcag ggagagctga agcaactgca ggagacccgg gcccaggatc
     2581 tggcctccaa ccagtcggaa tgtgacatgg cttgggtcaa gagagttggg gatgatcagg
     2641 tgaacttggc gctggcctac acagaactga cagaggagct gggtcggctt cgggagctga
     2701 gttccctgca ggggaggatc ttgaggactt tgctgcagga acaggcccgc aacgcaggat
     2761 ccggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2821 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2881 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2941 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3001 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3061 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg ggctgcaggt
     3121 cgaggcggat ctgatcaaga gacaggatga gcatcgatac catggtgagc aagggcgagg
     3181 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     3241 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     3301 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     3361 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     3421 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     3481 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     3541 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     3601 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     3661 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     3721 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     3781 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     3841 ccgccgggat cactctcggc atggacgagc tgtacaagta agatgcatcg catcgccttc
     3901 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgataaa ataaaagatt
     3961 ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta
     4021 gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat agagaagttc
     4081 agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat atctgtggta
     4141 agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata tgggccaaac
     4201 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
     4261 atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
     4321 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
     4381 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggggcgcc
     4441 agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt
     4501 gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc
     4561 gtcagcgggg gtctttcatt tgggggctcg tccgggatcg ggagacccct gcccagggac
     4621 caccgaccca ccaccgggag gtaagctggc tgcctcgcgc gtttcggtga tgacggtgaa
     4681 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     4741 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
     4801 acccagtcac gtagcgatag cggagtgtat ctagctaggt agctagagga tctttgtgaa
     4861 ggaaccttac ttctgtggtg tgacataatt ggacaaacta cctacagaga tttaaagctc
     4921 taaggtaaat ataaaatttt taagtgtata atgtgttaaa ctactgattc taattgtttg
     4981 tgtattttag attccaacct atggaactga tgaatgggag cagtggtgga atgcctttaa
     5041 tgaggaaaac ctgttttgct cagaagaaat gccatctagt gatgatgagg ctactgctga
     5101 ctctcaacat tctactcctc caaaaaagaa gagaaaggta gaagacccca aggactttcc
     5161 ttcagaattg ctaagttttt tgagtcatgc tgtgtttagt aatagaactc ttgcttgctt
     5221 tgctatttac accacaaagg aaaaagctgc actgctatac aagaaaatta tggaaaaata
     5281 tttgatgtat agtgccttga ctagagatca taatcagcca taccacattt gtagaggttt
     5341 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     5401 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5461 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5521 tcaatgtatc ttatcatgtc tggatcagct tcagaagatg gcggagggcc tccaacacag
     5581 taattttcct cccgactctt aaaatagaaa atgtcaagtc agttaagcag gaagtgacta
     5641 actgaccgca gctggccgtg cgacatcctc ttttaattag ttgctaggca actgccctcc
     5701 agagggcagt gtggttttgc aagaggaagc aaaaagcctc tccacccagg cctagaatgt
     5761 ttccacccaa tcattactat gacaacagct gtttttttta gtattaagca gaggccgggg
     5821 gcccctggcc tccgcttact ctggagaaaa agaagagagg cattgtagag gcttccagag
     5881 gcaacttgtc aaaacaggac tgcttctatt tctgtcacac tgtctggccc tgtcacaagg
     5941 tccagcacct ccataccccc tttaataagc agtttgggaa cgggtgcggg tcttactccg
     6001 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     6061 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     6121 ggaggctttt ttggaggctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6181 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6241 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6301 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6361 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6421 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6481 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6541 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6601 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6661 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6721 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6781 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     6841 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6901 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6961 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7021 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7081 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7141 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7201 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7261 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7321 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7381 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7441 ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7501 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7561 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7621 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7681 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7741 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7801 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7861 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7921 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7981 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8041 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8101 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     8161 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     8221 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     8281 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     8341 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     8401 gatgcgtaag gagaaaatac cgcatcaggc gcca
//
