LOCUS       sequence-492696-        8107 bp    DNA     circular SYN 12-NOV-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 Nov 12, 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"
     CDS             complement(177..188)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             381..614
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    686..807
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(723..742)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     777..796
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      847..982
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1014..1030)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1014..1030)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1027..1049)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1038..1054
                     /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(1062..1068,1069..1086,1087..1092))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1107..1128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1245..1262)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1416..2004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1496..1515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2175..2966,2967..3035))
                     /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     2798..2817
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3036..3140)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3208..3226
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3264..3286)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3386..3405
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3599..3621
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3613..3630
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3614..3630
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3645..3871
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             3872..4052
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4099..4224
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4717..4950
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5135..5179
                     /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    5446..5562
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        5585..5888
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5889..6092
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6043..6063
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6089..6113
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             6208..6231
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    join(7813..8107,1..294)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7866..7886)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
       61 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
      121 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
      181 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
      241 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
      301 ttcgagctcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
      361 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
      421 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
      481 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
      541 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
      601 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
      661 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
      721 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
      781 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
      841 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
      901 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
      961 tgaggaggct tttttggagg cctaggcttt tgcgggccca aattcgtaat catgtcatag
     1021 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     1081 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     1141 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1201 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     1261 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1321 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1381 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1441 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1501 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1561 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1621 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1681 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1741 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     1801 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     1861 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     1921 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     1981 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2041 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2101 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2161 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2221 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2281 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     2341 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     2401 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     2461 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     2521 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     2581 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     2641 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     2701 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     2761 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     2821 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     2881 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     2941 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3001 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3061 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3121 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     3181 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     3241 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     3301 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     3361 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     3421 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
     3481 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     3541 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     3601 cagtcacgac gttgtaaaac gacggccagt gccaagctga cgcgtgtagt cttatgcaat
     3661 actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa
     3721 aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag
     3781 gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat
     3841 tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc
     3901 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     3961 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     4021 acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg
     4081 aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca
     4141 agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag
     4201 gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa
     4261 aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca
     4321 agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt
     4381 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
     4441 ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc
     4501 aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa
     4561 gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
     4621 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
     4681 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
     4741 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     4801 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     4861 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     4921 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     4981 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     5041 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     5101 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     5161 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     5221 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     5281 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     5341 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
     5401 cagatccatt cgattagtga acggatctcg acggtatcgg ttaactttta aaagaaaagg
     5461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     5521 aactaaagaa ttacaaaaac aaattacaaa attcaaaatt ttatcgataa gcttgggagt
     5581 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     5641 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     5701 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     5761 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     5821 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     5881 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     5941 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     6001 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     6061 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga
     6121 gacgccatcc acgctgtttt gacctccata gaagacaccg actctagaac ctgcagcaag
     6181 ctttgatcag ctagcctcga gaccatggac tacaaggatg acgatgacaa ggaattcatg
     6241 agtctgctaa actgtgaaaa cagctgtgga tccagccagt ctgaaagtga ctgctgtgtg
     6301 gccatggcca gctcctgtag cgctgtaaca aaagatgata gtgtgggtgg aactgccagc
     6361 acggggaacc tctccagctc atttatggag gagatccagg gatatgatgt agagtttgac
     6421 ccacccctgg aaagcaagta tgaatgcccc atctgcttga tggcattacg agaagcagtg
     6481 caaacgccat gcggccatag gttctgcaaa gcctgcatca taaaatcaat aagggatgca
     6541 ggtcacaaat gtccagttga caatgaaata ctgctggaaa atcaactatt tccagacaat
     6601 tttgcaaaac gtgagattct ttctctgatg gtgaaatgtc caaatgaagg ttgtttgcac
     6661 aagatggaac tgagacatct tgaggatcat caagcacatt gtgagtttgc tcttatggat
     6721 tgtccccaat gccagcgtcc cttccaaaaa ttccatatta atattcacat tctgaaggat
     6781 tgtccaagga gacaggtttc ttgtgacaac tgtgctgcat caatggcatt tgaagataaa
     6841 gagatccatg accagaactg tcctttggca aatgtcatct gtgaatactg caatactata
     6901 ctcatcagag aacagatgcc taatcattat gatctagact gccctacagc cccaattcca
     6961 tgcacattca gtacttttgg ttgccatgaa aagatgcaga ggaatcactt ggcacgccac
     7021 ctacaagaga acacccagtc acacatgaga atgttggccc aggctgttca tagtttgagc
     7081 gttatacccg actctgggta tatctcagag gtccggaatt tccaggaaac tattcaccag
     7141 ttagagggtc gccttgtaag acaagaccat caaatccggg agctgactgc taaaatggaa
     7201 actcagagta tgtatgtaag tgagctcaaa cgaaccattc gaacccttga ggacaaagtt
     7261 gctgaaatcg aagcacagca gtgcaatgga atttatattt ggaagattgg caactttgga
     7321 atgcatttga aatgtcaaga agaggagaaa cctgttgtga ttcatagccc tggattctac
     7381 actggcaaac ccgggtacaa actgtgcatg cgcttgcacc ttcagttacc gactgctcag
     7441 cgctgtgcaa actatatatc cctttttgtc cacacaatgc aaggagaata tgacagccac
     7501 ctcccttggc ccttccaggg tacaatacgc cttacaattc ttgatcagtc tgaagcacct
     7561 gtaaggcaaa accacgaaga gataatggat gccaaaccag agctgcttgc tttccagcga
     7621 cccacaatcc cacggaaccc aaaaggtttt ggctatgtaa cttttatgca tctggaagcc
     7681 ctaagacaaa gaactttcat taaggatgac acattattag tgcgctgtga ggtctccacc
     7741 cgctttgaca tgggtagcct tcggagggag ggttttcagc cacgaagtac tgatgcaggg
     7801 gtataggtcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     7861 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     7921 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     7981 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     8041 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     8101 ttccccc
//
