LOCUS       sequence-190154-        7058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of TRAF3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7058)
  TITLE     Karin Lab plasmids
REFERENCE   2  (bases 1 to 7058)
  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: pcDNA3 HA-humanTRAF3 (1098)
FEATURES             Location/Qualifiers
     source          1..7058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        178..557
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        558..761
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     712..732
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     806..825
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        806..824
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             862..888
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             862..888
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2554..2572)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2555..2572)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2592..2609)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2598..2822
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2868..3296
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2955..2974)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3165..3186
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3305..3325)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3310..3639
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3490..3625
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3552..3571
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3706..4500
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3760..3779)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4370..4389
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4674..4795
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4711..4730)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4765..4784
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4844..4860)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4844..4860)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4857..4879)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4868..4884
                     /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(4892..4898,4899..4916,4917..4922))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4937..4958
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5075..5092)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5246..5834)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5326..5345)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6005..6796,6797..6865))
                     /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     6628..6647
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6866..6970)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(7045..7058,1..6))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
       61 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      121 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      181 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      241 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      301 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      361 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      421 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      481 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      541 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      601 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      661 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      721 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac
      781 ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc caagcttggt
      841 accgagctcg gatccaccat gtacccctac gacgtgcccg actacgccga attcatggag
      901 tcgagtaaaa agatggactc tcctggcgcg ctgcagacta acccgccgct aaagctgcac
      961 actgaccgca gtgctgggac gccagttttt gtccctgaac aaggaggtta caaggaaaag
     1021 tttgtgaaga ccgtggagga caagtacaag tgtgagaagt gccacctggt gctgtgcagc
     1081 ccgaagcaga ccgagtgtgg gcaccgcttc tgcgagagct gcatggcggc cctgctgagc
     1141 tcttcaagtc caaaatgtac agcgtgtcaa gagagcatcg ttaaagataa ggtgtttaag
     1201 gataattgct gcaagagaga aattctggct cttcagatct attgtcggaa tgaaagcaga
     1261 ggttgtgcag agcagttaat gctgggacat ctgctggtgc atttaaaaaa tgattgccat
     1321 tttgaagaac ttccatgtgt gcgtcctgac tgcaaagaaa aggtcttgag gaaagacctg
     1381 cgagaccacg tggagaaggc gtgtaaatac cgggaagcca catgcagcca ctgcaagagt
     1441 caggttccga tgatcgcgct gcagaaacac gaagacaccg actgtccctg cgtggtggtg
     1501 tcctgccctc acaagtgcag cgtccagact ctcctgagga gcgaggggac aaaccagcag
     1561 atcaaggccc acgaggccag ctccgccgtg cagcacgtca acctgctgaa ggagtggagc
     1621 aactcgctcg aaaagaaggt ttccttgttg cagaatgaaa gtgtagaaaa aaacaagagc
     1681 atacaaagtt tgcacaatca gatatgtagc tttgaaattg aaattgagag acaaaaggaa
     1741 atgcttcgaa ataatgaatc caaaatcctt catttacagc gagtgataga cagccaagca
     1801 gagaaactga aggagcttga caaggagatc cggcccttcc ggcagaactg ggaggaagca
     1861 gacagcatga agagcagcgt ggagtccctc cagaaccgcg tgaccgagct ggagagcgtg
     1921 gacaagagcg cggggcaagt ggctcggaac acaggcctgc tggagtccca gctgagccgg
     1981 catgaccaga tgctgagtgt gcacgacatc cgcctagccg acatggacct gcgcttccag
     2041 gtcctggaga ccgccagcta caatggagtg ctcatctgga agattcgcga ctacaagcgg
     2101 cggaagcagg aggccgtcat ggggaagacc ctgtcccttt acagccagcc tttctacact
     2161 ggttactttg gctataagat gtgtgccagg gtctacctga acggggacgg gatggggaag
     2221 gggacgcact tgtcgctgtt ttttgtcatc atgcgtggag aatatgatgc cctgcttcct
     2281 tggccgttta agcagaaagt gacactcatg ctgatggatc aggggtcctc tcgacgtcat
     2341 ttgggagatg cattcaagcc cgaccccaac agcagcagct tcaagaagcc cactggagag
     2401 atgaatatcg cctctggctg cccagtcttt gtggcccaaa ctgttctaga aaatgggaca
     2461 tatattaaag atgatacaat ttttattaaa gtcatagtgg atacttcgga tctgcccgat
     2521 ccctgactcg agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc
     2581 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2641 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2701 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2761 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2821 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     2881 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2941 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3001 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3061 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3121 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3181 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3241 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3301 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3361 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3421 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3481 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3541 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3601 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3661 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3721 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3781 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3841 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3901 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3961 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4021 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4081 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4141 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4201 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4261 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4321 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4381 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4441 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4501 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4561 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4621 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4681 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4741 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4801 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4861 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4921 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4981 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5041 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5101 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5161 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5221 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5281 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5341 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5401 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5461 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5521 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5581 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5641 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5701 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5761 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5821 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5881 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5941 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6001 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6061 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6121 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6181 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6241 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6301 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6361 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6421 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6481 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6541 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6601 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6661 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6721 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6781 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6841 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6901 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6961 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     7021 cgatccccta tggtgcactc tcagtacaat ctgctctg
//
