LOCUS       sequence-190145-        7089 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 7089)
  TITLE     Karin Lab plasmids
REFERENCE   2  (bases 1 to 7089)
  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 Flag-humanTRAF3 (1096)
FEATURES             Location/Qualifiers
     source          1..7089
                     /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             905..928
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2585..2603)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2586..2603)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2623..2640)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2629..2853
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2899..3327
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2986..3005)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3196..3217
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3336..3356)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3341..3670
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3521..3656
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3583..3602
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3737..4531
                     /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(3791..3810)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4401..4420
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4705..4826
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4742..4761)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4796..4815
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4875..4891)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4875..4891)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4888..4910)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4899..4915
                     /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(4923..4929,4930..4947,4948..4953))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4968..4989
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5106..5123)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5277..5865)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5357..5376)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6036..6827,6828..6896))
                     /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     6659..6678
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6897..7001)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(7076..7089,1..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 gatccactag taacggccgc cagtgtgctg gaattcgccc ttatgtctag
      901 aatggactac aaggacgacg atgacaagga gtcgagtaaa aagatggact ctcctggcgc
      961 gctgcagact aacccgccgc taaagctgca cactgaccgc agtgctggga cgccagtttt
     1021 tgtccctgaa caaggaggtt acaaggaaaa gtttgtgaag accgtggagg acaagtacaa
     1081 gtgtgagaag tgccacctgg tgctgtgcag cccgaagcag accgagtgtg ggcaccgctt
     1141 ctgcgagagc tgcatggcgg ccctgctgag ctcttcaagt ccaaaatgta cagcgtgtca
     1201 agagagcatc gttaaagata aggtgtttaa ggataattgc tgcaagagag aaattctggc
     1261 tcttcagatc tattgtcgga atgaaagcag aggttgtgca gagcagttaa tgctgggaca
     1321 tctgctggtg catttaaaaa atgattgcca ttttgaagaa cttccatgtg tgcgtcctga
     1381 ctgcaaagaa aaggtcttga ggaaagacct gcgagaccac gtggagaagg cgtgtaaata
     1441 ccgggaagcc acatgcagcc actgcaagag tcaggttccg atgatcgcgc tgcagaaaca
     1501 cgaagacacc gactgtccct gcgtggtggt gtcctgccct cacaagtgca gcgtccagac
     1561 tctcctgagg agcgagggga caaaccagca gatcaaggcc cacgaggcca gctccgccgt
     1621 gcagcacgtc aacctgctga aggagtggag caactcgctc gaaaagaagg tttccttgtt
     1681 gcagaatgaa agtgtagaaa aaaacaagag catacaaagt ttgcacaatc agatatgtag
     1741 ctttgaaatt gaaattgaga gacaaaagga aatgcttcga aataatgaat ccaaaatcct
     1801 tcatttacag cgagtgatag acagccaagc agagaaactg aaggagcttg acaaggagat
     1861 ccggcccttc cggcagaact gggaggaagc agacagcatg aagagcagcg tggagtccct
     1921 ccagaaccgc gtgaccgagc tggagagcgt ggacaagagc gcggggcaag tggctcggaa
     1981 cacaggcctg ctggagtccc agctgagccg gcatgaccag atgctgagtg tgcacgacat
     2041 ccgcctagcc gacatggacc tgcgcttcca ggtcctggag accgccagct acaatggagt
     2101 gctcatctgg aagattcgcg actacaagcg gcggaagcag gaggccgtca tggggaagac
     2161 cctgtccctt tacagccagc ctttctacac tggttacttt ggctataaga tgtgtgccag
     2221 ggtctacctg aacggggacg ggatggggaa ggggacgcac ttgtcgctgt tttttgtcat
     2281 catgcgtgga gaatatgatg ccctgcttcc ttggccgttt aagcagaaag tgacactcat
     2341 gctgatggat caggggtcct ctcgacgtca tttgggagat gcattcaagc ccgaccccaa
     2401 cagcagcagc ttcaagaagc ccactggaga gatgaatatc gcctctggct gcccagtctt
     2461 tgtggcccaa actgttctag aaaatgggac atatattaaa gatgatacaa tttttattaa
     2521 agtcatagtg gatacttcgg atctgcccga tccctgactc gagcatgcat ctagagggcc
     2581 ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta
     2641 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2701 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2761 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2821 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     2881 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2941 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3001 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3061 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3121 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3181 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3241 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3301 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     3361 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3421 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3481 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3541 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3601 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3661 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     3721 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3781 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3841 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3901 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3961 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4021 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4081 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4141 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4201 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4261 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4321 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4381 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4441 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4501 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4561 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4621 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4681 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     4741 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4801 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     4861 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4921 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4981 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5041 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5101 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5161 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5221 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5281 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5341 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5401 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5461 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5521 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5581 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5641 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5701 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5761 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5821 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5881 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5941 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6001 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6061 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6121 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6181 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6241 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6301 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6361 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6421 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6481 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6541 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6601 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6661 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6721 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6781 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6841 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6901 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6961 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7021 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     7081 tctgctctg
//
