LOCUS       sequence-433250-        6476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RIPLET lacking N-terminal RING domain in pcDNA3.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6476)
  AUTHORS   Cadena C, Ahmad S, Xavier A, Willemsen J, Park S, Park JW, Oh SW, 
            Fujita T, Hou F, Binder M, Hur S
  TITLE     Ubiquitin-Dependent and -Independent Roles of E3 Ligase RIPLET in 
            Innate Immunity.
  JOURNAL   Cell. 2019 May 16;177(5):1187-1200.e16. doi: 
            10.1016/j.cell.2019.03.017. Epub 2019 Apr 18.
  PUBMED    31006531
REFERENCE   2  (bases 1 to 6476)
  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: 
            "10.1016/j.cell.2019.03.017"; journalName: "Cell"; date: 
            "2019-05-16- 16"; volume: "177"; issue: "5"; pages: "1187-1200.e16"
COMMENT     Sequence Label: HA-RIPLET Delta-RING
FEATURES             Location/Qualifiers
     source          1..6476
                     /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             907..933
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2029..2047)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2030..2047)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2067..2084)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2073..2297
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2343..2771
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2430..2449)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2640..2661
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2780..2800)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2785..3114
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2965..3100
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3027..3046
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3181..3975
                     /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(3235..3254)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3845..3864
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4149..4270
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4186..4205)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4240..4259
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4319..4335)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4319..4335)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4332..4354)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4343..4359
                     /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(4367..4373,4374..4391,4392..4397))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4412..4433
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4550..4567)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4721..5309)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4801..4820)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5480..6271,6272..6340))
                     /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     6103..6122
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6341..6445)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 atggcgtacc catacgatgt tccagattac gctggatccg agatacaggc gggctccgac
      961 cctgcccact gcccctgccc gggctccagt tccctctcca gcgcggccgc gaggccccgg
     1021 cgccgcccgg aactgcagcg ggtggcagta gagaagagca tcacagaagt tgctcaggag
     1081 ctgacagagc tggtggaaca tcttgtagac attgtcagaa gcctgcagaa tcagaggccc
     1141 ctatcagaat ctggaccaga caacgaactg agcatcctgg gcaaggcttt ttcttctggg
     1201 gtggatcttt ccatggcttc tccaaagctg gtgacttccg acacagctgc agggaaaatc
     1261 agagatattc tccatgacct agaagaaatt caggaaaaat tacaagaaag cgtcacctgg
     1321 aaagaggctc ctgaagcaca aatgcaggga gaactcctgg aagccccgtc ttcctcctca
     1381 tgcccattgc ctgaccagag ccaccctgca ctcaggagag cttctcggtt tgctcagtgg
     1441 gccatccatc caacctttaa cttgaagagc ctttcctgca gcctggaggt gtccaaggat
     1501 tcccgtacag tgactgtgtc tcaccgccca caaccctatc gctggagctg tgagaggttt
     1561 tctaccagcc aggtcttatg ttcccaggcc ctgtcttctg gaaagcatta ctgggaagtg
     1621 gacactagga attgcagcca ctgggcagtt ggggtggctt cctgggagat gagccgcgac
     1681 caggtcctgg gaaggactat ggactcttgt tgtgtggaat ggaaggggac tagccagctc
     1741 tctgcatggc acatggtcaa ggaaactgtc cttggctcag acagacctgg ggtggtgggc
     1801 atctggctga accttgagga gggaaagctt gccttctatt cagtggacaa tcaggagaag
     1861 cttctgtatg agtgtaccat ctctgcctcc tctcctttgt accctgcctt ctggctgtat
     1921 ggcttacatc ctggaaatta cctgataata aagcaagtaa aggtgtaaga attctgcaga
     1981 tatccatcac actggcggcc gctcgagcat gcatctagag ggccctattc tatagtgtca
     2041 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2101 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2161 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2221 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2281 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     2341 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2401 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2461 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2521 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2581 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2641 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2701 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2761 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2821 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2881 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2941 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3001 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     3061 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     3121 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     3181 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3241 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3301 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3361 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3421 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3481 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3541 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3601 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3661 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     3721 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     3781 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     3841 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     3901 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     3961 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4021 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4081 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4141 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4201 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4261 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4321 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4381 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4441 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4501 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4561 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4621 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4681 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4741 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4801 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4861 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4921 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4981 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5041 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5101 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5161 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5221 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5281 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5341 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5401 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5461 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5521 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5581 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5641 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5701 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5761 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5821 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5881 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5941 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6001 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6061 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6121 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6181 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6241 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6301 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6361 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6421 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
