LOCUS       sequence-243301-        8774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of mouse Tbkbp1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8774)
  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 8774)
  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
FEATURES             Location/Qualifiers
     source          1..8774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        101..480
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        481..684
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     635..655
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             685..860
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    923..1122
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1323..1739
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1669..1691
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1707..1723
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1755..1781
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3136..3156)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3141..3470
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3321..3456
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(3529..3531,3532..3534,3535..4248)
                     /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(3529..3531,3532..3534,3535..4245)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3574..3595)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3835..3854)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4182..4203
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4354..4947
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     intron          5292..5357
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5487..5507
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5779..5913
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5816..5835)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5870..5889
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6371..6505
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6570..6587)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6741..7329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6821..6840)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7500..8291,8292..8360))
                     /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     8123..8142
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8361..8465)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8533..8551
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     8711..8730
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
       61 gcgatcggtg cgggcctctt cgctattacg ccagcgcgtt gacattgatt attgactagt
      121 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      181 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      241 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      301 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      361 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      421 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      481 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      541 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      601 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      661 tgggaggtct atataagcag agctcaataa aagagcccac aacccctcac tcggcgcgcc
      721 agtcttccga tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt
      781 tgcatccgaa tcgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc
      841 cacgacgggg gtctttcatt tgggggctcg tccgggattt ggagacccct gcccagggac
      901 caccgaccca ccaccgggag gtaagctggc cagcaactta tctgtgtctg tccgattgtc
      961 tagtgtctat gtttgatgtt atgcgcctgc gtctgtacta gttagctaac tagctctgta
     1021 tctggcggac ccgtggtgga actgacgagt tctgaacacc cggccgcaac cctgggagac
     1081 gtcccaggga ctttgggggc cgtttttgtg gcccgacctg aggaagggag tcgatgtgga
     1141 atccgacccc gtcaggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc
     1201 ctccgtctga atttttgctt tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc
     1261 gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt
     1321 agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc
     1381 ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg
     1441 cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca
     1501 tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc
     1561 cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg
     1621 tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc
     1681 ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccg
     1741 gaattatcat ggcctaccct tatgatgtgc cagattatgc ctctcccgaa ttcatggagt
     1801 ccatgtttga agatgacatc agcatcctga cccaagaggc cttggggccc agtgaggtgt
     1861 ggctggatgg tcctggagac ccgtccctgg gaggggacat gtgttctgcc tcccactttg
     1921 ccctcatcac agcctacgga gacatcaagg aaaggctagg aggcctggag agggagaatg
     1981 ccaccctccg ccgtcgcctc aaagtctatg agatcaagta tccgctgatc actgattttg
     2041 gagaggagca tggcttccct ctgtatgaac tcaaggatgg ctccctgctg gaggtggaga
     2101 aggtcagtct gcagcaacga ctcaaccaat tccaacacga gttgcagaag agtaaggagc
     2161 aggaggagca gctcggggag atgatccagg cttacgagaa actgtgcgtg gagaaaagcg
     2221 acttggagac agagctgggg gagatgcggg ccctggtaga gactcatcta cggcagatct
     2281 gtggtctgga gaaacagctg cagcaacagc agggcctccg ggacgcagct ttctccagcc
     2341 tgagcccccc agctgtcccc gcctcagcct gtcctgactt ggacctgcac tatctggctc
     2401 tgagaggggg acctgccttg ggtcatggtt ggcctggccc tacgagtgta agtgtgagcg
     2461 agctggagag gaggcgcctg gaagaggccc tggaggctgc gcagggggag gctcgggggg
     2521 ctcagctccg ggaggagcaa ctccaggcgg aatgcgagag acttcaggga gagctgaagc
     2581 aactgcagga gacccgggcc caggatctgg cctccaacca gtcggaatgt gacatggctt
     2641 gggtcaagag agttggggat gatcaggtga acttggcgct ggcctacaca gaactgacag
     2701 aggagctggg tcggcttcgg gagctgagtt ccctgcaggg gaggatcttg aggactttgc
     2761 tgcaggaaca ggcccgcaac gcaggccaaa ggcactcacc gctgtcgcaa cgtcactccc
     2821 cggcccctgc gtgcccctca ccctccccgc ctgcccgacc gccgccctgt gccccgtgcc
     2881 agtcccccgc tgcacagcgc cgttcgcccg tgcccccgtg cccttcgccc cagcagcgcc
     2941 gctcgcccgc ctcgccgtcc tgcccgtccc cagtcccaca gcgccgctcg cccgtgccgc
     3001 cgtcgtgcca gtcgcccagt cctcagcgcc gctcgcctct gccagctggg tttccctgtt
     3061 gggtacccag atgatgctct catcaaacac atagactccc accttgagaa cagcaagatc
     3121 tagggatccg gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3181 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3241 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3301 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3361 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3421 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttgggc
     3481 tgcaggtcga ggcggatctg atcaagagac aggatgagca tcgataccat ggtgagcaag
     3541 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     3601 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     3661 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     3721 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     3781 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     3841 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     3901 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     3961 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     4021 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     4081 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     4141 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     4201 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaga tgcatcgcat
     4261 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgataaaata
     4321 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     4381 caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga
     4441 gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc
     4501 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg
     4561 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     4621 tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     4681 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     4741 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     4801 gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct
     4861 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     4921 actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga gacccctgcc
     4981 cagggaccac cgacccacca ccgggaggta agctggctgc ctcgcgcgtt tcggtgatga
     5041 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     5101 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     5161 agccatgacc cagtcacgta gcgatagcgg agtgtatcta gctaggtagc tagaggatct
     5221 ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct acagagattt
     5281 aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta ctgattctaa
     5341 ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag tggtggaatg
     5401 cctttaatga ggaaaacctg ttttgctcag aagaaatgcc atctagtgat gatgaggcta
     5461 ctgctgactc tcaacattct actcctccaa aaaagaagag aaaggtagaa gaccccaagg
     5521 actttccttc agaattgcta agttttttga gtcatgctgt gtttagtaat agaactcttg
     5581 cttgctttgc tatttacacc acaaaggaaa aagctgcact gctatacaag aaaattatgg
     5641 aaaaatattt gatgtatagt gccttgacta gagatcataa tcagccatac cacatttgta
     5701 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     5761 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5821 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5881 aaactcatca atgtatctta tcatgtctgg atcagcttca gaagatggcg gagggcctcc
     5941 aacacagtaa ttttcctccc gactcttaaa atagaaaatg tcaagtcagt taagcaggaa
     6001 gtgactaact gaccgcagct ggccgtgcga catcctcttt taattagttg ctaggcaact
     6061 gccctccaga gggcagtgtg gttttgcaag aggaagcaaa aagcctctcc acccaggcct
     6121 agaatgtttc cacccaatca ttactatgac aacagctgtt ttttttagta ttaagcagag
     6181 gccgggggcc cctggcctcc gcttactctg gagaaaaaga agagaggcat tgtagaggct
     6241 tccagaggca acttgtcaaa acaggactgc ttctatttct gtcacactgt ctggccctgt
     6301 cacaaggtcc agcacctcca tacccccttt aataagcagt ttgggaacgg gtgcgggtct
     6361 tactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6421 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     6481 gtagtgagga ggcttttttg gaggctgcat taatgaatcg gccaacgcgc ggggagaggc
     6541 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6601 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6661 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6721 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6781 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6841 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6901 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6961 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     7021 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     7081 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7141 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     7201 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7261 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7321 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7381 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7441 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7501 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7561 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7621 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7681 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7741 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7801 gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7861 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7921 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7981 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     8041 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     8101 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     8161 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     8221 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     8281 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8341 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8401 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8461 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     8521 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     8581 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     8641 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     8701 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     8761 ccgcacagat gcgt
//
