LOCUS       sequence-190221-        6950 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of TRAF6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6950)
  TITLE     Karin Lab plasmids
REFERENCE   2  (bases 1 to 6950)
  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 humanTRAF6 (1090)
FEATURES             Location/Qualifiers
     source          1..6950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(49..68)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        240..619
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        620..823
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     774..794
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     868..887
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        868..886
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2508..2526)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2509..2526)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2546..2563)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2552..2776
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2822..3250
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2909..2928)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3119..3140
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3259..3279)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3264..3593
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3444..3579
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3506..3525
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3660..4454
                     /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(3714..3733)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4324..4343
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4628..4749
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4665..4684)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4719..4738
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4798..4814)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4798..4814)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4811..4833)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4822..4838
                     /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(4846..4852,4853..4870,4871..4876))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4891..4912
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5029..5046)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5200..5788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5280..5299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5959..6750,6751..6819))
                     /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     6582..6601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6820..6924)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
       61 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
      121 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
      181 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
      241 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      301 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      361 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      421 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      481 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      541 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      601 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      661 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      721 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      781 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
      841 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagcttg
      901 gtaccgaatt catgagtctg ctaaactgtg aaaacagctg tggatccagc cagtctgaaa
      961 gtgactgctg tgtggccatg gccagctcct gtagcgctgt aacaaaagat gatagtgtgg
     1021 gtggaactgc cagcacgggg aacctctcca gctcatttat ggaggagatc cagggatatg
     1081 atgtagagtt tgacccaccc ctggaaagca agtatgaatg ccccatctgc ttgatggcat
     1141 tacgagaagc agtgcaaacg ccatgcggcc ataggttctg caaagcctgc atcataaaat
     1201 caataaggga tgcaggtcac aaatgtccag ttgacaatga aatactgctg gaaaatcaac
     1261 tatttccaga caattttgca aaacgtgaga ttctttctct gatggtgaaa tgtccaaatg
     1321 aaggttgttt gcacaagatg gaactgagac atcttgagga tcatcaagca cattgtgagt
     1381 ttgctcttat ggattgtccc caatgccagc gtcccttcca aaaattccat attaatattc
     1441 acattctgaa ggattgtcca aggagacagg tttcttgtga caactgtgct gcatcaatgg
     1501 catttgaaga taaagagatc catgaccaga actgtccttt ggcaaatgtc atctgtgaat
     1561 actgcaatac tatactcatc agagaacaga tgcctaatca ttatgatcta gactgcccta
     1621 cagccccaat tccatgcaca ttcagtactt ttggttgcca tgaaaagatg cagaggaatc
     1681 acttggcacg ccacctacaa gagaacaccc agtcacacat gagaatgttg gcccaggctg
     1741 ttcatagttt gagcgttata cccgactctg ggtatatctc agaggtccgg aatttccagg
     1801 aaactattca ccagttagag ggtcgccttg taagacaaga ccatcaaatc cgggagctga
     1861 ctgctaaaat ggaaactcag agtatgtatg taagtgagct caaacgaacc attcgaaccc
     1921 ttgaggacaa agttgctgaa atcgaagcac agcagtgcaa tggaatttat atttggaaga
     1981 ttggcaactt tggaatgcat ttgaaatgtc aagaagagga gaaacctgtt gtgattcata
     2041 gccctggatt ctacactggc aaacccgggt acaaactgtg catgcgcttg caccttcagt
     2101 taccgactgc tcagcgctgt gcaaactata tatccctttt tgtccacaca atgcaaggag
     2161 aatatgacag ccacctccct tggcccttcc agggtacaat acgccttaca attcttgatc
     2221 agtctgaagc acctgtaagg caaaaccacg aagagataat ggatgccaaa ccagagctgc
     2281 ttgctttcca gcgacccaca atcccacgga acccaaaagg ttttggctat gtaactttta
     2341 tgcatctgga agccctaaga caaagaactt tcattaagga tgacacatta ttagtgcgct
     2401 gtgaggtctc cacccgcttt gacatgggta gccttcggag ggagggtttt cagccacgaa
     2461 gtactgatgc aggggtatag ctcgagcatg catctagagg gccctattct atagtgtcac
     2521 ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2581 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2641 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2701 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2761 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2821 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2881 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2941 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3001 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3061 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3121 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3181 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3241 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3301 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3361 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3421 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3481 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3541 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3601 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     3661 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3721 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3781 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3841 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3901 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3961 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4021 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4081 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4141 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4201 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4261 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4321 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4381 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4441 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4501 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4561 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4621 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4681 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4741 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4801 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4861 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4921 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4981 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5041 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5101 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5161 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5221 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5281 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5341 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5401 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5461 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5521 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5581 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5641 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5701 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5761 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5821 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5881 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5941 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6001 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6061 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6121 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6181 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6241 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6301 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6361 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6421 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6481 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6541 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6601 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6661 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6721 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6781 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6841 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6901 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
