LOCUS       sequence-433252-        6161 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RIPLET lacking C-terminal PRY/SPRY domain in pcDNA3.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6161)
  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 6161)
  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-PRY/SPRY
FEATURES             Location/Qualifiers
     source          1..6161
                     /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(1714..1732)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1715..1732)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1752..1769)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1758..1982
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2028..2456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2115..2134)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2325..2346
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2465..2485)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2470..2799
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2650..2785
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2712..2731
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2866..3660
                     /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(2920..2939)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3530..3549
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3834..3955
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3871..3890)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3925..3944
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4004..4020)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4004..4020)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4017..4039)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4028..4044
                     /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(4052..4058,4059..4076,4077..4082))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4097..4118
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4235..4252)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4406..4994)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4486..4505)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5165..5956,5957..6025))
                     /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     5788..5807
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6026..6130)
                     /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 gctggatcca tggcgggcct gggcctgggc
      961 tccgccgttc ccgtgtggct ggccgaggac gacctcggct gcatcatctg ccaggggctg
     1021 ctggactggc ccgccacgct gccctgcggc cacagcttct gccgccactg cctggaggcc
     1081 ctgtggggcg cccgcgacgc ccgccgctgg gcctgcccca cttgccgcca gggcgccgcg
     1141 cagcagccgc acctgcggaa gaacacgcta ctgcaggacc tggccgacaa gtaccgccgc
     1201 gccgcacgcg agatacaggc gggctccgac cctgcccact gcccctgccc gggctccagt
     1261 tccctctcca gcgcggccgc gaggccccgg cgccgcccgg aactgcagcg ggtggcagta
     1321 gagaagagca tcacagaagt tgctcaggag ctgacagagc tggtggaaca tcttgtagac
     1381 attgtcagaa gcctgcagaa tcagaggccc ctatcagaat ctggaccaga caacgaactg
     1441 agcatcctgg gcaaggcttt ttcttctggg gtggatcttt ccatggcttc tccaaagctg
     1501 gtgacttccg acacagctgc agggaaaatc agagatattc tccatgacct agaagaaatt
     1561 caggaaaaat tacaagaaag cgtcacctgg aaagaggctc ctgaagcaca aatgcaggga
     1621 gaactcctgg aagccccgtc ttcctcctca taagaattct gcagatatcc atcacactgg
     1681 cggccgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc
     1741 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1801 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1861 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     1921 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     1981 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     2041 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2101 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2161 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2221 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2281 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2341 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2401 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2461 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2521 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2581 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2641 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2701 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     2761 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     2821 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2881 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2941 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3001 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3061 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3121 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3181 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3241 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3301 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3361 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     3421 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3481 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3541 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3601 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3661 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3721 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3781 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     3841 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3901 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3961 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4021 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4081 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4141 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4201 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4261 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4321 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4381 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4441 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4501 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4561 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4621 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4681 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4741 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4801 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4861 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4921 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4981 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5041 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5101 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5161 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5221 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5281 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5341 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5401 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5461 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5521 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5581 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5641 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5701 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5761 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5821 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5881 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5941 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6001 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6061 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6121 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
