LOCUS       sequence-243863-        8284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of mouse Tbkbp1 (1-280).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8284)
  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 8284)
  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 1-280
FEATURES             Location/Qualifiers
     source          1..8284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             91..507
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     437..459
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     475..491
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             523..549
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1414..1434)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1419..1748
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1599..1734
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(1807..1809,1810..1812,1813..2526)
                     /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(1807..1809,1810..1812,1813..2523)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1852..1873)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2113..2132)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2460..2481
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2632..3225
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     intron          3570..3635
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3765..3785
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4057..4191
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4094..4113)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4148..4167
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4649..4783
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4848..4865)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5019..5607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5099..5118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5778..6569,6570..6638))
                     /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     6401..6420
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6639..6743)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6811..6829
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     6989..7008
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7153..7532
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7533..7736
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7687..7707
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             7737..7912
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    7975..8174
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg
       61 actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc cttaagtttg
      121 accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag
      181 aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg
      241 cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct
      301 ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt
      361 gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca
      421 tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat
      481 ccagccctca ctccttctct aggcgccgga attatcatgg cctaccctta tgatgtgcca
      541 gattatgcct ctcccgaatt catggagtcc atgtttgaag atgacatcag catcctgacc
      601 caagaggcct tggggcccag tgaggtgtgg ctggatggtc ctggagaccc gtccctggga
      661 ggggacatgt gttctgcctc ccactttgcc ctcatcacag cctacggaga catcaaggaa
      721 aggctaggag gcctggagag ggagaatgcc accctccgcc gtcgcctcaa agtctatgag
      781 atcaagtatc cgctgatcac tgattttgga gaggagcatg gcttccctct gtatgaactc
      841 aaggatggct ccctgctgga ggtggagaag gtcagtctgc agcaacgact caaccaattc
      901 caacacgagt tgcagaagag taaggagcag gaggagcagc tcggggagat gatccaggct
      961 tacgagaaac tgtgcgtgga gaaaagcgac ttggagacag agctggggga gatgcgggcc
     1021 ctggtagaga ctcatctacg gcagatctgt ggtctggaga aacagctgca gcaacagcag
     1081 ggcctccggg acgcagcttt ctccagcctg agccccccag ctgtccccgc ctcagcctgt
     1141 cctgacttgg acctgcacta tctggctctg agagggggac ctgccttggg tcatggttgg
     1201 cctggcccta cgagtgtaag tgtgagcgag ctggagagga ggcgcctgga agaggccctg
     1261 gaggctgcgc agggggaggc tcggggggct cagctccggg aggagcaact ccaggcggaa
     1321 tgcgagagac ttcagggaga gctgaagcaa ctgcaggaga cccgggccca ggatctggcc
     1381 tccaaccagt cggaatgtga cggatccggc tgtggaatgt gtgtcagtta gggtgtggaa
     1441 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     1501 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     1561 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     1621 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     1681 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     1741 tttgcaaaaa gcttgggctg caggtcgagg cggatctgat caagagacag gatgagcatc
     1801 gataccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     1861 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     1921 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     1981 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     2041 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     2101 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2161 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2221 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     2281 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     2341 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     2401 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     2461 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     2521 aagtaagatg catcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     2581 ctggggttcg ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
     2641 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa
     2701 tacataactg agaatagaga agttcagatc aaggtcagga acagatggaa cagctgaata
     2761 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     2821 tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     2881 agggccaaga acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca
     2941 tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac
     3001 caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag
     3061 cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt
     3121 gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg
     3181 gtctcctctg agtgattgac tacccgtcag cgggggtctt tcatttgggg gctcgtccgg
     3241 gatcgggaga cccctgccca gggaccaccg acccaccacc gggaggtaag ctggctgcct
     3301 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     3361 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     3421 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatctagc
     3481 taggtagcta gaggatcttt gtgaaggaac cttacttctg tggtgtgaca taattggaca
     3541 aactacctac agagatttaa agctctaagg taaatataaa atttttaagt gtataatgtg
     3601 ttaaactact gattctaatt gtttgtgtat tttagattcc aacctatgga actgatgaat
     3661 gggagcagtg gtggaatgcc tttaatgagg aaaacctgtt ttgctcagaa gaaatgccat
     3721 ctagtgatga tgaggctact gctgactctc aacattctac tcctccaaaa aagaagagaa
     3781 aggtagaaga ccccaaggac tttccttcag aattgctaag ttttttgagt catgctgtgt
     3841 ttagtaatag aactcttgct tgctttgcta tttacaccac aaaggaaaaa gctgcactgc
     3901 tatacaagaa aattatggaa aaatatttga tgtatagtgc cttgactaga gatcataatc
     3961 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     4021 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     4081 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4141 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat cagcttcaga
     4201 agatggcgga gggcctccaa cacagtaatt ttcctcccga ctcttaaaat agaaaatgtc
     4261 aagtcagtta agcaggaagt gactaactga ccgcagctgg ccgtgcgaca tcctctttta
     4321 attagttgct aggcaactgc cctccagagg gcagtgtggt tttgcaagag gaagcaaaaa
     4381 gcctctccac ccaggcctag aatgtttcca cccaatcatt actatgacaa cagctgtttt
     4441 ttttagtatt aagcagaggc cgggggcccc tggcctccgc ttactctgga gaaaaagaag
     4501 agaggcattg tagaggcttc cagaggcaac ttgtcaaaac aggactgctt ctatttctgt
     4561 cacactgtct ggccctgtca caaggtccag cacctccata ccccctttaa taagcagttt
     4621 gggaacgggt gcgggtctta ctccgcccat cccgccccta actccgccca gttccgccca
     4681 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4741 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggctgcatta atgaatcggc
     4801 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4861 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4921 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4981 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5041 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5101 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5161 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5221 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5281 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5341 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5401 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5461 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5521 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5581 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5641 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5701 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5761 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5821 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5881 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5941 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6001 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6061 gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg
     6121 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6181 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6241 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6301 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6361 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6421 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6481 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6541 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6601 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6661 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6721 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     6781 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     6841 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     6901 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     6961 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     7021 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     7081 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     7141 cgccagcgcg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     7201 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     7261 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     7321 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     7381 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     7441 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     7501 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     7561 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     7621 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     7681 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcaat
     7741 aaaagagccc acaacccctc actcggcgcg ccagtcttcc gatagactgc gtcgcccggg
     7801 tacccgtatt cccaataaag cctcttgctg tttgcatccg aatcgtggtc tcgctgttcc
     7861 ttgggagggt ctcctctgag tgattgacta cccacgacgg gggtctttca tttgggggct
     7921 cgtccgggat ttggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg
     7981 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgtttgatg ttatgcgcct
     8041 gcgtctgtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
     8101 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
     8161 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
     8221 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
     8281 gaac
//
