LOCUS       sequence-149201-        6885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To overexpress mRipk3 in Mammalian Cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6885)
  AUTHORS   Seo J, Lee EW, Sung H, Seong D, Dondelinger Y, Shin J, Jeong M, Lee 
            HK, Kim JH, Han SY, Lee C, Seong JK, Vandenabeele P, Song J
  TITLE     CHIP controls necroptosis through ubiquitylation- and 
            lysosome-dependent degradation of RIPK3.
  JOURNAL   Nat Cell Biol. 2016 Mar;18(3):291-302. doi: 10.1038/ncb3314. Epub 
            2016 Feb 22.
  PUBMED    26900751
REFERENCE   2  (bases 1 to 6885)
  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: "10.1038/ncb3314"; 
            journalName: "Nat Cell Biol"; date: "2016-03"; volume: "18"; issue: 
            "3"; pages: "291-302"
COMMENT     Sequence Label: pcDNA3-HA-mRipk3
FEATURES             Location/Qualifiers
     source          1..6885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             910..936
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2298..2309
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(2437..2455)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2438..2455)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2475..2492)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2481..2705
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2751..3179
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2838..2857)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3048..3069
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3188..3208)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3193..3523
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3374..3509
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3436..3455
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3590..4384
                     /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(3644..3663)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4254..4273
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4558..4679
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4595..4614)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4649..4668
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4728..4744)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4728..4744)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4741..4763)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4752..4768
                     /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(4776..4782,4783..4800,4801..4806))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4821..4842
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4959..4976)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5130..5718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5210..5229)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5889..6680,6681..6749))
                     /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     6512..6531
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6750..6854)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 atggcctcct acccttatga tgtgccagat tatgcctctc ccgaattcat gtcttctgtc
      961 aagttatggc ctactggtgc gtcagcggtt cctctggtga gccgtgaaga actgaagaag
     1021 ctggagtttg tgggcaaagg agggttcgga gtcgtgttcc gggcacacca cagaacatgg
     1081 aaccatgatg tagcagtcaa gatcgtgaac tcgaagaaga tatcctggga ggtgaaggct
     1141 atggttaatc ttcgtaatga gaacgttctg ctcctgctgg gggtcactga ggacctccag
     1201 tgggacttcg tgtccgggca ggctctggtg acaagattca tggagaatgg ctccctcgca
     1261 gggctgctgc aacccgagtg ccctcggccc tggccactcc tctgtcgcct gctgcaggaa
     1321 gtggtgctgg ggatgtgcta cctacacagc ttgaaccctc cgctcctgca ccgggacctc
     1381 aagccctcta acattctgct ggatccagag ctccacgcca agctagcaga ttttggcctg
     1441 tccacgtttc agggagggtc ccagtcaggg tcaggatcag gatcaggatc cagggactct
     1501 gggggcaccc tagcgtactt ggacccagag ctgttattta atgtcaacct gaaggcttct
     1561 aaagcgagtg atgtctacag ctttgggatc ctcgtgtggg cagtgctggc tggcagagaa
     1621 gctgagttgg tagacaagac ttcactaatc cgggaaacag tgtgtgacag gcagagtcgt
     1681 cctccactga cagagctgcc tccaggtagc cctgagactc ctggcttgga aaaactgaag
     1741 gagttaatga ttcattgctg gggttcccag tccgaaaaca ggccatcctt ccaggactgc
     1801 gaaccaaaaa ccaatgaagt ttacaatctg gtaaaggaca aggtagatgc tgctgtctcc
     1861 gaggtaaagc attatctgtc tcagcacaga agcagcggca gaaacttgtc tgccagagag
     1921 ccaagccaaa gaggcacaga aatggattgc ccgagggaaa ccatggtttc taaaatgctg
     1981 gaccgcctgc atttggagga accctccgga ccagttcctg gaaaatgtcc tgagaggcaa
     2041 gcacaggaca catcagttgg gcctgccaca ccagcaagga catcttctga ccccgtggct
     2101 ggcactcctc agattccaca tactttaccc ttcagaggca caacacctgg gccagtcttt
     2161 actgagactc ccggtcctca cccccaaagg aatcagggag atggaagaca cggcactcct
     2221 tggtatccct ggaccccacc gaatccaatg acagggccac cggctctcgt cttcaacaac
     2281 tgttctgaag tgcagattgg gaactacaac tccttggtag caccaccaag aactactgcc
     2341 tcaagttcgg ccaagtatga ccaagcacag ttcggcaggg gtaggggctg gcagcccttc
     2401 cacaagtagc tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag
     2461 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2521 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2581 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2641 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     2701 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     2761 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2821 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2881 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     2941 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3001 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3061 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3121 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3181 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc aggcaggcag
     3241 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3301 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3361 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3421 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     3481 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     3541 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     3601 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3661 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3721 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     3781 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3841 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3901 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3961 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4021 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4081 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     4141 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4201 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4261 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4321 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4381 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4441 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4501 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     4561 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4621 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4681 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     4741 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4801 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4861 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4921 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4981 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5041 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5101 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5161 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5221 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5281 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     5341 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5401 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5461 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5521 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5581 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5641 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5701 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5761 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5821 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5881 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5941 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6001 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6061 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6121 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6181 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6241 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6301 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6361 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6421 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6481 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6541 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6601 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6661 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6721 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6781 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6841 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
