LOCUS       sequence-495756-        8107 bp    DNA     circular SYN 18-DEC-2025
DEFINITION  Lentiviral Expression of TRAF6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8107)
  TITLE     Karin Lab plasmids
REFERENCE   2  (bases 1 to 8107)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pLU-CMV-PL3 Flag-humanTRAF6-WT (1111)
FEATURES             Location/Qualifiers
     source          1..8107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             624..647
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2229..2817
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2282..2302)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2700..2711)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2904..3137
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3209..3330
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3246..3265)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3300..3319
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3370..3505
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3537..3553)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3537..3553)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3550..3572)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3561..3577
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3585..3591,3592..3609,3610..3615))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3630..3651
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3768..3785)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3939..4527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4019..4038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4698..5489,5490..5558))
                     /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     5321..5340
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5559..5663)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5731..5749
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5787..5809)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5909..5928
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6122..6144
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6136..6153
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6137..6153
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6168..6394
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6395..6575
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6622..6747
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7240..7473
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7658..7702
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7969..8085
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgactc tagaacctgc agcaagcttt
      601 gatcagctag cctcgagacc atggactaca aggatgacga tgacaaggaa ttcatgagtc
      661 tgctaaactg tgaaaacagc tgtggatcca gccagtctga aagtgactgc tgtgtggcca
      721 tggccagctc ctgtagcgct gtaacaaaag atgatagtgt gggtggaact gccagcacgg
      781 ggaacctctc cagctcattt atggaggaga tccagggata tgatgtagag tttgacccac
      841 ccctggaaag caagtatgaa tgccccatct gcttgatggc attacgagaa gcagtgcaaa
      901 cgccatgcgg ccataggttc tgcaaagcct gcatcataaa atcaataagg gatgcaggtc
      961 acaaatgtcc agttgacaat gaaatactgc tggaaaatca actatttcca gacaattttg
     1021 caaaacgtga gattctttct ctgatggtga aatgtccaaa tgaaggttgt ttgcacaaga
     1081 tggaactgag acatcttgag gatcatcaag cacattgtga gtttgctctt atggattgtc
     1141 cccaatgcca gcgtcccttc caaaaattcc atattaatat tcacattctg aaggattgtc
     1201 caaggagaca ggtttcttgt gacaactgtg ctgcatcaat ggcatttgaa gataaagaga
     1261 tccatgacca gaactgtcct ttggcaaatg tcatctgtga atactgcaat actatactca
     1321 tcagagaaca gatgcctaat cattatgatc tagactgccc tacagcccca attccatgca
     1381 cattcagtac ttttggttgt catgaaaaga tgcagaggaa tcacttggca cgccacctac
     1441 aagagaacac ccagtcacac atgagaatgt tggcccaggc tgttcatagt ttgagcgtta
     1501 tacccgactc tgggtatatc tcagaggtcc ggaatttcca ggaaactatt caccagttag
     1561 agggtcgcct tgtaagacaa gaccatcaaa tccgggagct gactgctaaa atggaaactc
     1621 agagtatgta tgtaagtgag ctcaaacgaa ccattcgaac ccttgaggac aaagttgctg
     1681 aaatcgaagc acagcagtgc aatggaattt atatttggaa gattggcaac tttggaatgc
     1741 atttgaaatg tcaagaagag gagaaacctg ttgtgattca tagccctgga ttctacactg
     1801 gcaaacccgg gtacaaactg tgcatgcgct tgcaccttca gttaccgact gctcagcgct
     1861 gtgcaaacta tatatccctt tttgtccaca caatgcaagg agaatatgac agccacctcc
     1921 cttggccctt ccagggtaca atacgcctta caattcttga tcagtctgaa gcacctgtaa
     1981 ggcaaaacca cgaagagata atggatgcca aaccagagct gcttgctttc cagcgaccca
     2041 caatcccacg gaacccaaaa ggttttggct atgtaacttt tatgcatctg gaagccctaa
     2101 gacaaagaac tttcattaag gatgacacat tattagtgcg ctgtgaggtc tccacccgct
     2161 ttgacatggg tagccttcgg agggagggtt ttcagccacg aagtactgat gcaggggtat
     2221 aggtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2281 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2341 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     2401 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     2461 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     2521 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     2581 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     2641 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     2701 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     2761 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     2821 gaattcgagc tcggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     2881 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
     2941 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     3001 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     3061 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     3121 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     3181 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     3241 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3301 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     3361 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3421 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3481 tagtgaggag gcttttttgg aggcctaggc ttttgcgggc ccaaattcgt aatcatgtca
     3541 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3601 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3661 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3721 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3781 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3841 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3901 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3961 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4021 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4081 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4141 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4201 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4261 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4321 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4381 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4441 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4501 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4561 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4621 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4681 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4741 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4801 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4861 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4921 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4981 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5041 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5101 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5161 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5221 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5281 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5341 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5401 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5461 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5521 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5581 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5641 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     5701 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     5761 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     5821 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     5881 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     5941 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     6001 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     6061 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     6121 tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tgacgcgtgt agtcttatgc
     6181 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
     6241 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
     6301 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
     6361 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
     6421 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     6481 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     6541 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
     6601 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
     6661 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
     6721 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
     6781 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
     6841 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
     6901 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     6961 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     7021 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     7081 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     7141 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     7201 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     7261 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     7321 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     7381 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     7441 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     7501 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     7561 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     7621 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     7681 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     7741 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     7801 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     7861 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     7921 agacagatcc attcgattag tgaacggatc tcgacggtat cggttaactt ttaaaagaaa
     7981 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     8041 acaaactaaa gaattacaaa aacaaattac aaaattcaaa attttatcga taagcttggg
     8101 agttccg
//
