LOCUS       sequence-416263-        6741 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To detect localization of RIPK1 protein in live mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6741)
  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 6741)
  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: pEGFP-N2-RIPK1
FEATURES             Location/Qualifiers
     source          1..6741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2621..2630
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2627..2629,2630..2632,2633..3346)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2627..2629,2630..2632,2633..3343)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2672..2693)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2933..2952)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3280..3301
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3469..3590
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3506..3525)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3560..3579
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3597..4052)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3734..3755)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3946..3965
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4079..4183
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4185..4542
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4185..4376
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4208..4228)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4393..4528
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4577..5371
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4577..5371
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4631..4650)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5241..5260
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5559..5578)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5603..5650
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5979..6567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6468..6487
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagaa tgcaaccaga catgtccttg aatgtcatta agatgaaatc
      601 cagtgacttc ctggagagtg cagaactgga cagcggaggc tttgggaagg tgtctctgtg
      661 tttccacaga acccagggac tcatgatcat gaaaacagtg tacaaggggc ccaactgcat
      721 tgagcacaac gaggccctct tggaggaggc gaagatgatg aacagactga gacacagccg
      781 ggtggtgaag ctcctgggcg tcatcataga ggaagggaag tactccctgg tgatggagta
      841 catggagaag ggcaacctga tgcacgtgct gaaagccgag atgagtactc cgctttctgt
      901 aaaaggaagg ataattttgg aaatcattga aggaatgtgc tacttacatg gaaaaggcgt
      961 gatacacaag gacctgaagc ctgaaaatat ccttgttgat aatgacttcc acattaagat
     1021 cgcagacctc ggccttgcct cctttaagat gtggagcaaa ctgaataatg aagagcacaa
     1081 tgagctgagg gaagtggacg gcaccgctaa gaagaatggc ggcaccctct actacatggc
     1141 gcccgagcac ctgaatgacg tcaacgcaaa gcccacagag aagtcggatg tgtacagctt
     1201 tgctgtagta ctctgggcga tatttgcaaa taaggagcca tatgaaaatg ctatctgtga
     1261 gcagcagttg ataatgtgca taaaatctgg gaacaggcca gatgtggatg acatcactga
     1321 gtactgccca agagaaatta tcagtctcat gaagctctgc tgggaagcga atccggaagc
     1381 tcggccgaca tttcctggca ttgaagaaaa atttaggcct ttttatttaa gtcaattaga
     1441 agaaagtgta gaagaggacg tgaagagttt aaagaaagag tattcaaacg aaaatgcagt
     1501 tgtgaagaga atgcagtctc ttcaacttga ttgtgtggca gtaccttcaa gccggtcaaa
     1561 ttcagccaca gaacagcctg gttcactgca cagttcccag ggacttggga tgggtcctgt
     1621 ggaggagtcc tggtttgctc cttccctgga gcacccacaa gaagagaatg agcccagcct
     1681 gcagagtaaa ctccaagacg aagccaacta ccatctttat ggcagccgca tggacaggca
     1741 gacgaaacag cagcccagac agaatgtggc ttacaacaga gaggaggaaa ggagacgcag
     1801 ggtctcccat gacccttttg cacagcaaag accttacgag aattttcaga atacagaggg
     1861 aaaaggcact gcttattcca gtgcagccag tcatggtaat gcagtgcacc agccctcagg
     1921 gctcaccagc caacctcaag tactgtatca gaacaatgga ttatatagct cacatggctt
     1981 tggaacaaga ccactggatc caggaacagc aggtcccaga gtttggtaca ggccaattcc
     2041 aagtcatatg cctagtctgc ataatatccc agtgcctgag accaactatc taggaaatac
     2101 acccaccatg ccattcagct ccttgccacc aacagatgaa tctataaaat ataccatata
     2161 caatagtact ggcattcaga ttggagccta caattatatg gagattggtg ggacgagttc
     2221 atcactacta gacagcacaa atacgaactt caaagaagag ccagctgcta agtaccaagc
     2281 tatctttgat aataccacta gtctgacgga taaacacctg gacccaatca gggaaaatct
     2341 gggaaagcac tggaaaaact gtgcccgtaa actgggcttc acacagtctc agattgatga
     2401 aattgaccat gactatgagc gagatggact gaaagaaaag gtttaccaga tgctccaaaa
     2461 gtgggtgatg agggaaggca taaagggagc cacggtgggg aagctggccc aggcgctcca
     2521 ccagtgttcc aggatcgacc ttctgagcag cttgatttac gtcagccaga acgaattctg
     2581 cagtcgacgg taccgcgggc ccgggatcca ccggccggtc gccaccatgg tgagcaaggg
     2641 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2701 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2761 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2821 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     2881 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     2941 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     3001 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     3061 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     3121 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     3181 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     3241 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     3301 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc
     3361 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     3421 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     3481 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3541 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     3601 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3661 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3721 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3781 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3841 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3901 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3961 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4021 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     4081 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4141 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     4201 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4261 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4321 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4381 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4441 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4501 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     4561 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4621 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4681 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4741 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4801 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4861 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4921 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4981 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5041 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5101 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5161 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5221 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5281 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5341 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     5401 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5461 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5521 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     5581 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     5641 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     5701 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     5761 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     5821 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5881 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5941 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6001 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6061 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6121 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6181 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6241 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6301 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6361 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6421 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6481 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6541 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6601 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6661 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
     6721 tagcccatat atggagttcc g
//
