LOCUS       sequence-190249-        8008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentiviral Expression of TRAF2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8008)
  TITLE     Karin Lab plasmids
REFERENCE   2  (bases 1 to 8008)
  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"
COMMENT     Sequence Label: pLU-CMV-PL3 Flag-humanTRAF2-WT (1108)
FEATURES             Location/Qualifiers
     source          1..8008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     165..187
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     179..196
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     180..196
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        211..437
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             438..618
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    665..790
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1283..1516
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1701..1745
                     /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    2012..2128
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2151..2454
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2455..2658
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2609..2629
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2655..2679
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2741..2764
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    4280..4868
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4333..4353)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4751..4762)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4955..5188
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5260..5381
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5297..5316)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5351..5370
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5421..5556
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5588..5604)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5588..5604)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5601..5623)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5612..5628
                     /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(5636..5642,5643..5660,5661..5666))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5681..5702
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5819..5836)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5990..6578)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6070..6089)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6749..7540,7541..7609))
                     /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     7372..7391
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7610..7714)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7782..7800
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7838..7860)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7960..7979
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
       61 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      121 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      181 taaaacgacg gccagtgcca agctgacgcg tgtagtctta tgcaatactc ttgtagtctt
      241 gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat
      301 gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc
      361 tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc
      421 tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg
      481 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
      541 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
      601 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
      661 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
      721 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
      781 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
      841 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
      901 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
      961 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     1021 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     1081 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
     1141 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
     1201 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
     1261 gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa
     1321 gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta
     1381 tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac
     1441 tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa
     1501 aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg
     1561 tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct
     1621 ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag
     1681 aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa
     1741 gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga
     1801 tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag
     1861 ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg
     1921 acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat
     1981 tagtgaacgg atctcgacgg tatcggttaa cttttaaaag aaaagggggg attggggggt
     2041 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
     2101 aaaaacaaat tacaaaattc aaaattttat cgataagctt gggagttccg cgttacataa
     2161 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     2221 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     2281 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     2341 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     2401 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     2461 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     2521 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     2581 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     2641 gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
     2701 tgttttgacc tccatagaag acaccgactc tagaaccatg gactacaagg atgacgatga
     2761 caaggctgca gctagcgtga ccccccctgg ctccctggag ttgctacagc ccggcttctc
     2821 caagaccctc ctggggacca agctggaagc caagtacctg tgctccgcct gcagaaacgt
     2881 cctccgcagg cccttccagg cgcagtgtgg ccaccggtac tgctccttct gcctggccag
     2941 catcctcagc tctgggcctc agaactgtgc tgcctgtgtt cacgagggca tatatgaaga
     3001 aggcatttct attttagaaa gcagttcggc cttcccagat aatgctgccc gcagggaggt
     3061 ggagagcctg ccggccgtct gtcccagtga tggatgcacc tggaagggga ccctgaaaga
     3121 atacgagagc tgccacgaag gccgctgccc gctcatgctg accgaatgtc ccgcgtgcaa
     3181 aggcctggtc cgccttggtg aaaaggagcg ccacctggag cacgagtgcc cggagagaag
     3241 cctgagctgc cggcattgcc gggcaccctg ctgcggagca gacgtgaagg cgcaccacga
     3301 ggtctgcccc aagttcccct taacttgtga cggctgcggc aagaagaaga tcccccggga
     3361 gaagtttcag gaccacgtca agacttgtgg caagtgtcga gtcccttgca gattccacgc
     3421 catcggctgc ctcgagacgg tagagggtga gaaacagcag gagcacgagg tgcagtggct
     3481 gcgggagcac ctggccatgc tactgagctc ggtgctggag gcaaagcccc tcttgggaga
     3541 ccagagccac gcggggtcag agctcctgca gaggtgcgag agcctggaga agaagacggc
     3601 cacttttgag aacattgtct gcgtcctgaa ccgggaggtg gagagggtgg ccatgactgc
     3661 cgaggcctgc agccggcagc accggctgga ccaagacaag attgaagccc tgagtagcaa
     3721 ggtgcagcag ctggagagga gcattggcct caaggacctg gcgatggctg acttggagca
     3781 gaaggtcttg gagatggagg catccaccta cgatggggtc ttcatctgga agatctcaga
     3841 cttcgccagg aagcgccagg aagctgtggc tggccgcata cccgccatct tctccccagc
     3901 cttctacacc agcaggtacg gctacaagat gtgtctgcgt atctacctga acggcgacgg
     3961 caccgggcga ggaacacacc tgtccctctt ctttgtggtg atgaagggcc cgaatgacgc
     4021 cctgctgcgg tggcccttca accagaaggt gaccttaatg ctgctcgacc agaataaccg
     4081 ggagcacgtg attgacgcct tcaggcccga cgtgacttca tcctcctttc agaggccagt
     4141 caacgacatg aacatcgcaa gcggctgccc cctcttctgc cccgtctcca agatggaggc
     4201 aaagaattcc tacgtgcggg acgatgccat cttcatcaag gccattgtgg acctgacagg
     4261 gctctaagga tccgtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     4321 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     4381 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     4441 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     4501 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     4561 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     4621 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     4681 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     4741 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     4801 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     4861 ctccccgcct ggaattcgag ctcggtacct ttaagaccaa tgacttacaa ggcagctgta
     4921 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     4981 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     5041 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     5101 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     5161 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
     5221 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     5281 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5341 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     5401 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5461 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     5521 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcggg cccaaattcg
     5581 taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5641 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5701 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5761 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5821 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5881 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5941 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6001 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6061 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6121 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6181 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6241 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6301 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6361 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6421 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6481 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6541 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6601 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6661 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6721 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6781 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6841 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6901 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6961 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7021 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7081 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7141 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7201 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7261 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7321 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7381 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7441 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7501 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7561 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7621 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7681 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7741 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     7801 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     7861 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     7921 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     7981 tgagagtgca ccatatgcgg tgtgaaat
//
