LOCUS       sequence-261301-        9162 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retroviral expression plasmid for RIPK1-wt.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9162)
  AUTHORS   Zhang L, Blackwell K, Workman LM, Chen S, Pope MR, Janz S, Habelhah 
            H
  TITLE     RIP1 Cleavage in the Kinase Domain Regulates TRAIL-Induced NF-kappaB
            Activation and Lymphoma Survival.
  JOURNAL   Mol Cell Biol. 2015 Oct;35(19):3324-38. doi: 10.1128/MCB.00692-15. 
            Epub 2015 Jul 20.
  PUBMED    26195820
REFERENCE   2  (bases 1 to 9162)
  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: 
            "10.1128/MCB.00692-15"; journalName: "Mol Cell Biol"; date: 
            "2015-10"; volume: "35"; issue: "19"; pages: "3324-38"
COMMENT     Sequence Label: pMIG-Flag-RIPK1-wt
FEATURES             Location/Qualifiers
     source          1..9162
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2211..2258)
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             complement(2281..2697)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2297..2313)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2329..2351)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    2764..3105
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             3169..3685
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(3885..3904)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4032..4054)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4248..4267)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4367..4389
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4427..4445)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4513..4617
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4618..4686,4687..5478)
                     /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(4836..4855)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5649..6237
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6138..6157
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6391..6408
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6525..6546
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6561..6566,6567..6584,6585..6591)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6599..6615
                     /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)."
     LTR             6777..7291
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7338..7926
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             7444..7455
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7853..7873
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(8036..8749,8750..8752,8753..8755))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8081..8102)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     8430..8449
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8689..8710
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    join(8767..9162,1..157)
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    join(8769..9162,1..175)
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8929..8948)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     join(9158..9162,1..13)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
ORIGIN
        1 cctttggcga gaggggaaag acccctagga atgctcgtca agaagacagg gccaggtttc
       61 cgggccctca cattgccaaa agacggcaat atggtggaaa ataacatata gacaaacgca
      121 caccggcctt attccaagcg gcttcggcca gtaacgttag gggggggggg gcggaattct
      181 tagttctggc tgacgtaaat caagctgctc agaaggtcga tcctggaaca ctggtggagc
      241 gcctgggcca gcttccccac cgtggctccc tttatgcctt ccctcatcac ccacttttgg
      301 agcatctggt aaaccttttc tttcagtcca tctcgctcat agtcatggtc aatttcatca
      361 atctgagact gtgtgaagcc cagtttacgg gcacagtttt tccagtgctt tcccagattt
      421 tccctgatcg ggtccaggtg tttatccgtc agactagtgg tattatcaaa gatagcttgg
      481 tacttagcag ctggctcttc tttgaagttc gtatttgtgc tgtctagtag tgatgaactc
      541 gtcccaccaa tctccatata attgtaggct ccaatctgaa tgccagtact attgtatatg
      601 gtatatttta tagattcatc tgttggtggc aaggagctga atggcatggt gggtgtattt
      661 cctagatagt tggtctcagg cactgggata ttatgcagac taggcatatg acttggaatt
      721 ggcctgtacc aaactctggg acctgctgtt cctggatcca gtggtcttgt tccaaagcca
      781 tgtgagctat ataatccatt gttctgatac agtacttgag gttggctggt gagccctgat
      841 ggctggtgca ctgcattacc atgactggct gcactggaat aagcagtgcc ttttccctct
      901 gtattctgaa aattctcgta aggtctttgc tgtgcaaaag ggtcatggga gaccctgcgc
      961 ctcctttcct cctctctgtt gtaagccaca ttctgtctgg gctgctgttt cgtctgcctg
     1021 tccatgcggc tgccataaag atggtagttg gcttcgtctt ggagtttact ctgcaggctg
     1081 ggctcattct cttcttgtgg gtgctccagg gaaggagcaa accaggactc ctccacagga
     1141 cccatcccaa gtccctggga actgtgcagt gaaccaggct gttctgtggc tgaatttgac
     1201 cggcttgaag gtactgccac acaatcaagt tgaagagact gcattctctt cacaactgca
     1261 ttttcgtttg aatactcttt ctttaaactc ttcacgtcct cttctacact ttcttctaat
     1321 tgacttaaat aaaaaggcct aaatttttct tcaatgccag gaaatgtcgg ccgagcttcc
     1381 ggattcgctt cccagcagag cttcatgaga ctgataattt ctcttgggca gtactcagtg
     1441 atgtcatcca catctggcct gttcccagat tttatgcaca ttatcaactg ctgctcacag
     1501 atagcatttt catatggctc cttatttgca aatatcgccc agagtactac agcaaagctg
     1561 tacacatccg acttctctgt gggctttgcg ttgacgtcat tcaggtgctc gggcgccatg
     1621 tagtagaggg tgccgccatt cttcttagcg gtgccgtcca cttccctcag ctcattgtgc
     1681 tcttcattat tcagtttgct ccacatctta aaggaggcaa ggccgaggtc tgcgatctta
     1741 atgtggaagt cattatcaac aaggatattt tcaggcttca ggtccttgtg tatcacgcct
     1801 tttccatgta agtagcacat tccttcaatg atttccaaaa ttatccttcc ttttacagaa
     1861 agcggagtac tcatctcggc tttcagcacg tgcatcaggt tgcccttctc catgtactcc
     1921 atcaccaggg agtactgccc ttcctctatg atgacgccca ggagcttcac cacccggctg
     1981 tgtctcagtc tgttcatcat cttcgcctcc tccaagaggg cctcgttgtg ctcaatgcag
     2041 ttgggcccct tgtacactgt tttcatgatc atgagtccct gggttctgtg gaaacacaga
     2101 gacaccttcc cgaagcctcc gctgtccagt tctgcactct ccaggaagtc actggatttc
     2161 atcttaatga cattcaagga catgtctggt tgcatagatc ctgggtgacc tttatcgtca
     2221 tcgtctttgt agtctttatc gtcatcgtct ttgtagtcca tggtggagag atctaattcc
     2281 ggcgcctaga gaaggagtga gggctggata aagggaggat cgaggcgggg tcgaacgagg
     2341 aggttcaagg gggagagacg gggcggatgg aggaagagga ggcggaggct tagggtgtac
     2401 aaagggcttg acccagggag gggggtcaaa agccaaggct tcccaggtca cgatgtaggg
     2461 gacctggtct gggtgtccat gcgggccagg tgaaaagacc ttgatcttaa cctgggtgat
     2521 gaggtctcgg ttaaaggtgc cgtctcgcgg ccatccgacg ttaaaggttg gccattctgc
     2581 agagcagaag gtaacccaac gtctcttctt gacatctacc gactggttgt gagcgatccg
     2641 ctcgacatct ttccagtgac ctaaggtcaa acttaaggga gtggtaacag tctggcccta
     2701 attttcagac aaatacagaa acacagtcag acagagacaa cacagaacga tgctgcagcg
     2761 ctgcagcaga caagacgcgc ggcttcggtt ccaaaccgaa agcaaaaatt cagacggagg
     2821 cgggaactgt tttaggttct cgtctcctac cagaaccaca tatcctgacg gggtcggatt
     2881 ccacatcgac tcccttcctc aggtcgggcc acaaaaacgg cccccaaagt ccctgggacg
     2941 tctcccaggg ttgcggccgg gtgttcagaa ctcgtcagtt ccaccacggg tccgccagat
     3001 acagagctag ttagctaact agtacagacg caggcgcaaa cattagatgc cggcacaaac
     3061 acgcacaaag acagagacag acacgaaacg accgctggcc agcttacctc ccggcggggg
     3121 ggtcggtggt ccctgggcag gggtctccaa atctcggtgg aacctccaaa tgaaagaccc
     3181 ccgaggtggg cagtcaatca atctgaggag accctcccaa ggatcagcga gtccacgatt
     3241 cggatgcaaa cagcaagagg ctttattggg aatacgggta cccgggcgac gcagtctatc
     3301 ggaggactgg cgcgccgagt gaggggttgt gggctctttt attgagctcg gggagcagaa
     3361 gcgcgcgaac agaagcgaga agcgaactga ttggttagtt caaataaggc acagggtcat
     3421 ttcaggtcct tggggcaccc tggaaacatc tgatggttct ctagaaactg ctgagggcgg
     3481 gaccgcatct ggggaccatc tgttcttggc cctgagccgg ggcaggaact gcttaccaca
     3541 gatatcctgt ttggcccata ttctgctgtc tctctgttcc taaccttgat ctgaacttct
     3601 ctattctcag ttatgtattt tccatgcctt gcaaaatggc gttacttaag ctagcttgcc
     3661 aaacctacag gtggggtctt tcattccccc ctttttctgg agactaaata aaatctttta
     3721 ttttatctat ggctcgtact ctataggctt cagctggtga tattgttgag tcaaaactag
     3781 agcctggacc actgatatcc tgtctttaac aaattggact aatcgcctct acgccggacg
     3841 catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat
     3901 caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg
     3961 tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt
     4021 ccttgcgccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     4081 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     4141 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     4201 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     4261 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4321 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4381 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4441 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4501 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4561 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4621 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4681 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4741 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4801 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4861 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4921 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4981 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5041 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5101 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5161 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5221 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5281 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5341 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5401 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5461 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5521 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5581 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5641 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5701 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5761 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5821 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5881 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5941 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6001 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6061 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6121 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6181 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6241 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6301 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6361 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     6421 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     6481 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     6541 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     6601 gtgagcggat aacaatttca caattctccc cagctctttc tgcgctgtcc ataatgaact
     6661 atttcaggag tattggagta tgtggattca aaacagtggc taattgagtc aggattactt
     6721 aatattcgac tcctaccctc agaatgttgt tctccaactt caagaaactg ttacccatga
     6781 aagacccccg ctgacgggta gtcaatcact cagaggagac cctcccaagg aacagcgaga
     6841 ccacaagtcg gatgcaactg caagagggtt tattggatac acgggtaccc gggcgacgca
     6901 gtctatcgga ggactggcgc gccgagtgag gggttgtggg ctcttttatt gagctcgggg
     6961 agcagaagcg cgcgaacaga agcgagaagc gaactgattg gttagttcaa ataaggcaca
     7021 gggtcatttc aggtccttgg ggcaccctgg aaacatctga tggttctcta gaaactgctg
     7081 agggcgggac cgcatctggg gaccatctgt tcttggccct gagccggggc aggaactgct
     7141 taccacagat atcctgtttg gcccatattc tgctgtctct ctgttcctaa ccttgatctg
     7201 aacttctcta ttctcagtta tgtattttcc atgccttgca aaatggcgtt acttaagcta
     7261 gcttgccaaa cctacaggtg gggtctttca ttcccccctt tttctggaga ctaaataaaa
     7321 tcttttattt tatcgatgcg gggaggcggc ccaaagggag atccgactcg tctgagggcg
     7381 aaggcgaaga cgcggaagag gccgcagagc cggcagcagg ccgcgggaag gaaggtccgc
     7441 tggattgagg gccgaaggga cgtagcagaa ggacgtcccg cgcagaatcc aggtggcaac
     7501 acaggcgagc agccaaggaa aggacgatga tttccccgac aacaccacgg aattgtcagt
     7561 gcccaacagc cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt
     7621 ggcaataggg agggggaaag cgaaagtccc ggaaaggagc tgacaggtgg tggcaatgcc
     7681 ccaaccagtg ggggttgcgt cagcaaacac agtgcacacc acgccacgtt gcctgacaac
     7741 gggccacaac tcctcataaa gagacagcaa ccaggattta tacaaggagg agaaaatgaa
     7801 agccatacgg gaagcaatag catgatacaa aggcattaaa gcagcgtatc cacatagcgt
     7861 aaaaggagca acatagttaa gaataccagt caatctttca caaattttgt aatccagagg
     7921 ttgattatcg ataagcttgg ctgcaggtcg actctagaaa gtgtctcatg ccggaggcca
     7981 ttaaagacag gccttactgg ctgaatagaa gcggtgataa ttcttaatta attcattact
     8041 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
     8101 tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctgggtgctc aggtagtggt
     8161 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
     8221 gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcgccttg atgccgttct
     8281 tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc
     8341 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
     8401 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga
     8461 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     8521 tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg
     8581 gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg
     8641 tggcatcgcc ctcgccctcg ccagacacgc tgaacttgtg gccgtttacg tcgccgtcca
     8701 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatggttg
     8761 tggccatatt atcatcgtgt ttttcaaagg aaaaccacgt ccccgtggtt cggggggcct
     8821 agacgttttt ttaacctcga ctaaacacat gtaaagcatg tgcaccgagg ccccagatca
     8881 gatcccatac aatggggtac cttctgggca tccttcagcc ccttgttgaa tacgcttgag
     8941 gagagccatt tgactctttc cacaactatc caactcacaa cgtggcactg gggttgtgcc
     9001 gcctttgcag gtgtatctta tacacgtggc ttttggccgc agaggcacct gtcgccaggt
     9061 ggggggttcc gctgcctgca aagggtcgct acagacgttg tttgtcttca agaagcttcc
     9121 agaggaactg cttccttcac gacattcaac agaccttgca tt
//
