LOCUS       sequence-324559-        6377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HA-tagged mouse TRAF5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6377)
  AUTHORS   Wang X, Jin C, Tang Y, Tang LY, Zhang YE
  TITLE     Ubiquitination of tumor necrosis factor receptor-associated factor 4
            (TRAF4) by Smad ubiquitination regulatory factor 1 (Smurf1) 
            regulates motility of breast epithelial and cancer cells.
  JOURNAL   J Biol Chem. 2013 Jul 26;288(30):21784-92. doi: 
            10.1074/jbc.M113.472704. Epub 2013 Jun 11.
  PUBMED    23760265
REFERENCE   2  (bases 1 to 6377)
  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"; doi: 
            "10.1074/jbc.M113.472704"; journalName: "J Biol Chem"; date: 
            "2013-07-26- 26"; volume: "288"; issue: "30"; pages: "21784-92"
COMMENT     Sequence Label: pRK-HA-TRAF5
FEATURES             Location/Qualifiers
     source          1..6377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             930..956
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             936..962
                     /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="YDVPDYASL"
     regulatory      2677..2686
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2690..2824
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2727..2746)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2781..2800
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2893..3250
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2893..3084
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2916..2936)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3101..3236
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3270..3287)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3270..3286)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3279..3301)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3499..3954
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3586..3605)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3796..3817
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3971..3990)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4090..4112
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4150..4168)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4236..4340
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4341..4409,4410..5201)
                     /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     complement(4559..4578)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5372..5960
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5861..5880
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6114..6131
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6248..6269
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6284..6289,6290..6307,6308..6314)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6322..6338
                     /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)."
     primer_bind     6327..6349
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6346..6362
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6346..6362
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgccgcc accatgtgct acccatacga tgttccagat tacgctagtc
      961 ttatcgatga attcggatcc atggctcatt cggaggagca agcggctgtc ccctgcgcct
     1021 tcatccgcca gaactctggc aactcaattt ccttggactt tgagcccgac accgagtacc
     1081 agtttgtgga gcagctggaa gaacgctaca aatgtgcctt ctgccactcc gtgcttcaca
     1141 acccccacca gaccggctgc gggcaccgct tctgccagca gtgcatccgg tctctgagag
     1201 aattgaactc ggtgccgatc tgcccggtag acaaggaggt catcaagcct caggaggtgt
     1261 tcaaagacaa ctgctgcaaa agagaagttc tcaatttaca cgtctactgc aaaaacgccc
     1321 ccgggtgcaa tgccaggatt attctgggac gattccagga ccaccttcag cactgttcct
     1381 tccaagccgt gccctgccct aacgagagct gccgggaagc catgctccgg aaagacgtga
     1441 aagagcacct gagcgcatac tgccggttcc gagaggagaa gtgcctttac tgcaaaaggg
     1501 acatagtggt gaccaacctg caggatcatg aggaaaactc gtgtcctgcg tacccagtgt
     1561 cttgtcccaa caggtgtgtg cagactattc caagagctag ggtgaatgaa caccttactg
     1621 tatgtcctga ggctgagcaa gactgtccct ttaagcacta tggctgcact gtcaagggta
     1681 agcgggggaa cttgctggag catgagcggg cagccctgca ggaccacatg cttctggttt
     1741 tagagaagaa ctaccaacta gaacagcgga tctctgattt atatcagagt ctcgaacaga
     1801 aggaaagcaa gatccagcag ctggcagaaa ccgtgaagaa gttcgaaaag gagcttaagc
     1861 agttcacaca gatgtttggc agaaatggaa ctttcctctc aaatgtccag gctctcacca
     1921 gtcacacgga caagtcagct tggctggaag cgcaggtgcg gcagctgcta caaatagtta
     1981 accagcagcc aagtcgactt gatctgaggt ctttggtgga tgcggttgac agcgtgaaac
     2041 agaggatcac ccagctggaa gccagtgacc agagattagt tcttttagag ggggagacca
     2101 gcaagcacga cgcacacatt aatatccaca aagcacagct gaataagaac gaagagcggt
     2161 ttaagcagct ggagggcgcc tgctacagtg gcaagctcat ctggaaggtg acagattaca
     2221 gggtgaagaa gagggaggcc gtggaggggc acacagtgtc cgtcttcagc cagcctttct
     2281 acaccagccg ctgcggctac cggctctgtg ccagggcgta cctgaacggg gacgggtcgg
     2341 ggaagggaac gcacctgtcc ctgtactttg tggtgatgcg cggtgagttt gactcgctgc
     2401 tgcagtggcc gttcaggcag agggtgaccc tgatgctttt ggaccagagc ggcaagaaga
     2461 accatattgt ggagaccttc aaagctgacc ccaacagcag cagcttcaaa aggccagatg
     2521 gcgagatgaa cattgcctct ggctgtcccc gctttgtgtc gcactctact ctggagaact
     2581 ccaagaacac ctacattaaa gacgacacac tgttcttgaa agtggccgtg gatttaactg
     2641 acttggagga tctgtagctc gacctgcaga agcttggccg ccatggccca acttgtttat
     2701 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2761 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     2821 gatcgggaat taattcggcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg
     2881 gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     2941 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3001 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3061 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3121 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3181 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3241 cttttgcaaa aagctgttaa cagcttggca ctggccgtcg ttttacaacg tcgtgactgg
     3301 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     3361 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     3421 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3481 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3541 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3601 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3661 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     3721 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3781 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3841 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3901 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     3961 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4021 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4081 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4141 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     4201 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4261 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4321 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4381 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4441 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4501 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4561 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4621 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4681 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4741 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4801 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4861 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4921 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4981 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5041 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5101 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5161 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5221 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5281 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5341 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5401 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5461 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5521 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5581 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5641 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5701 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5761 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5821 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5881 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5941 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6001 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6061 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     6121 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     6181 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     6241 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     6301 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     6361 acatgattac gaattaa
//
