LOCUS       sequence-199543-        9164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retroviral expression plasmid for RIPK1-D324A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9164)
  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 9164)
  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-D324A
FEATURES             Location/Qualifiers
     source          1..9164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2045..2092)
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             complement(2115..2531)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2131..2147)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2163..2185)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    2598..2939
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             3003..3519
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(3719..3738)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3866..3888)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4082..4101)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4201..4223
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4261..4279)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4347..4451
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4452..4520,4521..5312)
                     /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(4670..4689)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5483..6071
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5972..5991
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6225..6242
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6359..6380
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6395..6400,6401..6418,6419..6425)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6433..6449
                     /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             6611..7125
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7172..7760
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             7278..7289
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7687..7707
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(7870..8583,8584..8586,8587..8589))
                     /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(7915..7936)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     8264..8283
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8523..8544
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    8601..9153
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8763..8782)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     8992..9009
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
ORIGIN
        1 gggggcggaa ttcttagttc tggctgacgt aaatcaagct gctcagaagg tcgatcctgg
       61 aacactggtg gagcgcctgg gccagcttcc ccaccgtggc tccctttatg ccttccctca
      121 tcacccactt ttggagcatc tggtaaacct tttctttcag tccatctcgc tcatagtcat
      181 ggtcaatttc atcaatctga gactgtgtga agcccagttt acgggcacag tttttccagt
      241 gctttcccag attttccctg atcgggtcca ggtgtttatc cgtcagacta gtggtattat
      301 caaagatagc ttggtactta gcagctggct cttctttgaa gttcgtattt gtgctgtcta
      361 gtagtgatga actcgtccca ccaatctcca tataattgta ggctccaatc tgaatgccag
      421 tactattgta tatggtatat tttatagatt catctgttgg tggcaaggag ctgaatggca
      481 tggtgggtgt atttcctaga tagttggtct caggcactgg gatattatgc agactaggca
      541 tatgacttgg aattggcctg taccaaactc tgggacctgc tgttcctgga tccagtggtc
      601 ttgttccaaa gccatgtgag ctatataatc cattgttctg atacagtact tgaggttggc
      661 tggtgagccc tgatggctgg tgcactgcat taccatgact ggctgcactg gaataagcag
      721 tgccttttcc ctctgtattc tgaaaattct cgtaaggtct ttgctgtgca aaagggtcat
      781 gggagaccct gcgcctcctt tcctcctctc tgttgtaagc cacattctgt ctgggctgct
      841 gtttcgtctg cctgtccatg cggctgccat aaagatggta gttggcttcg tcttggagtt
      901 tactctgcag gctgggctca ttctcttctt gtgggtgctc cagggaagga gcaaaccagg
      961 actcctccac aggacccatc ccaagtccct gggaactgtg cagtgaacca ggctgttctg
     1021 tggctgaatt tgaccggctt gaaggtactg ccacacaagc aagttgaaga gactgcattc
     1081 tcttcacaac tgcattttcg tttgaatact ctttctttaa actcttcacg tcctcttcta
     1141 cactttcttc taattgactt aaataaaaag gcctaaattt ttcttcaatg ccaggaaatg
     1201 tcggccgagc ttccggattc gcttcccagc agagcttcat gagactgata atttctcttg
     1261 ggcagtactc agtgatgtca tccacatctg gcctgttccc agattttatg cacattatca
     1321 actgctgctc acagatagca ttttcatatg gctccttatt tgcaaatatc gcccagagta
     1381 ctacagcaaa gctgtacaca tccgacttct ctgtgggctt tgcgttgacg tcattcaggt
     1441 gctcgggcgc catgtagtag agggtgccgc cattcttctt agcggtgccg tccacttccc
     1501 tcagctcatt gtgctcttca ttattcagtt tgctccacat cttaaaggag gcaaggccga
     1561 ggtctgcgat cttaatgtgg aagtcattat caacaaggat attttcaggc ttcaggtcct
     1621 tgtgtatcac gccttttcca tgtaagtagc acattccttc aatgatttcc aaaattatcc
     1681 ttccttttac agaaagcgga gtactcatct cggctttcag cacgtgcatc aggttgccct
     1741 tctccatgta ctccatcacc agggagtact gcccttcctc tatgatgacg cccaggagct
     1801 tcaccacccg gctgtgtctc agtctgttca tcatcttcgc ctcctccaag agggcctcgt
     1861 tgtgctcaat gcagttgggc cccttgtaca ctgttttcat gatcatgagt ccctgggttc
     1921 tgtggaaaca cagagacacc ttcccgaagc ctccgctgtc cagttctgca ctctccagga
     1981 agtcactgga tttcatctta atgacattca aggacatgtc tggttgcata gatcctgggt
     2041 gacctttatc gtcatcgtct ttgtagtctt tatcgtcatc gtctttgtag tccatggtgg
     2101 agagatctaa ttccggcgcc tagagaagga gtgagggctg gataaaggga ggatcgaggc
     2161 ggggtcgaac gaggaggttc aagggggaga gacggggcgg atggaggaag aggaggcgga
     2221 ggcttagggt gtacaaaggg cttgacccag ggaggggggt caaaagccaa ggcttcccag
     2281 gtcacgatgt aggggacctg gtctgggtgt ccatgcgggc caggtgaaaa gaccttgatc
     2341 ttaacctggg tgatgaggtc tcggttaaag gtgccgtctc gcggccatcc gacgttaaag
     2401 gttggccatt ctgcagagca gaaggtaacc caacgtctct tcttgacatc taccgactgg
     2461 ttgtgagcga tccgctcgac atctttccag tgacctaagg tcaaacttaa gggagtggta
     2521 acagtctggc cctaattttc agacaaatac agaaacacag tcagacagag acaacacaga
     2581 acgatgctgc agcgctgcag cagacaagac gcgcggcttc ggttccaaac cgaaagcaaa
     2641 aattcagacg gaggcgggaa ctgttttagg ttctcgtctc ctaccagaac cacatatcct
     2701 gacggggtcg gattccacat cgactccctt cctcaggtcg ggccacaaaa acggccccca
     2761 aagtccctgg gacgtctccc agggttgcgg ccgggtgttc agaactcgtc agttccacca
     2821 cgggtccgcc agatacagag ctagttagct aactagtaca gacgcaggcg caaacattag
     2881 atgccggcac aaacacgcac aaagacagag acagacacga aacgaccgct ggccagctta
     2941 cctcccggcg ggggggtcgg tggtccctgg gcaggggtct ccaaatctcg gtggaacctc
     3001 caaatgaaag acccccgagg tgggcagtca atcaatctga ggagaccctc ccaaggatca
     3061 gcgagtccac gattcggatg caaacagcaa gaggctttat tgggaatacg ggtacccggg
     3121 cgacgcagtc tatcggagga ctggcgcgcc gagtgagggg ttgtgggctc ttttattgag
     3181 ctcggggagc agaagcgcgc gaacagaagc gagaagcgaa ctgattggtt agttcaaata
     3241 aggcacaggg tcatttcagg tccttggggc accctggaaa catctgatgg ttctctagaa
     3301 actgctgagg gcgggaccgc atctggggac catctgttct tggccctgag ccggggcagg
     3361 aactgcttac cacagatatc ctgtttggcc catattctgc tgtctctctg ttcctaacct
     3421 tgatctgaac ttctctattc tcagttatgt attttccatg ccttgcaaaa tggcgttact
     3481 taagctagct tgccaaacct acaggtgggg tctttcattc cccccttttt ctggagacta
     3541 aataaaatct tttattttat ctatggctcg tactctatag gcttcagctg gtgatattgt
     3601 tgagtcaaaa ctagagcctg gaccactgat atcctgtctt taacaaattg gactaatcgc
     3661 ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc
     3721 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct
     3781 tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc
     3841 ttgcatgcac cattccttgc gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     3901 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     3961 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga
     4021 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
     4081 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4141 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4201 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     4261 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4321 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4381 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4441 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4501 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4561 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4621 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4681 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4741 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4801 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4861 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4921 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4981 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5041 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5101 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5161 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5221 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5281 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5341 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5401 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5461 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5521 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5581 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5641 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5701 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5761 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5821 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5881 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5941 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6001 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6061 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6121 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6181 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6241 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     6301 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     6361 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     6421 tgttgtgtgg aattgtgagc ggataacaat ttcacaattc tccccagctc tttctgcgct
     6481 gtccataatg aactatttca ggagtattgg agtatgtgga ttcaaaacag tggctaattg
     6541 agtcaggatt acttaatatt cgactcctac cctcagaatg ttgttctcca acttcaagaa
     6601 actgttaccc atgaaagacc cccgctgacg ggtagtcaat cactcagagg agaccctccc
     6661 aaggaacagc gagaccacaa gtcggatgca actgcaagag ggtttattgg atacacgggt
     6721 acccgggcga cgcagtctat cggaggactg gcgcgccgag tgaggggttg tgggctcttt
     6781 tattgagctc ggggagcaga agcgcgcgaa cagaagcgag aagcgaactg attggttagt
     6841 tcaaataagg cacagggtca tttcaggtcc ttggggcacc ctggaaacat ctgatggttc
     6901 tctagaaact gctgagggcg ggaccgcatc tggggaccat ctgttcttgg ccctgagccg
     6961 gggcaggaac tgcttaccac agatatcctg tttggcccat attctgctgt ctctctgttc
     7021 ctaaccttga tctgaacttc tctattctca gttatgtatt ttccatgcct tgcaaaatgg
     7081 cgttacttaa gctagcttgc caaacctaca ggtggggtct ttcattcccc cctttttctg
     7141 gagactaaat aaaatctttt attttatcga tgcggggagg cggcccaaag ggagatccga
     7201 ctcgtctgag ggcgaaggcg aagacgcgga agaggccgca gagccggcag caggccgcgg
     7261 gaaggaaggt ccgctggatt gagggccgaa gggacgtagc agaaggacgt cccgcgcaga
     7321 atccaggtgg caacacaggc gagcagccaa ggaaaggacg atgatttccc cgacaacacc
     7381 acggaattgt cagtgcccaa cagccgagcc cctgtccagc agcgggcaag gcaggcggcg
     7441 atgagttccg ccgtggcaat agggaggggg aaagcgaaag tcccggaaag gagctgacag
     7501 gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa acacagtgca caccacgcca
     7561 cgttgcctga caacgggcca caactcctca taaagagaca gcaaccagga tttatacaag
     7621 gaggagaaaa tgaaagccat acgggaagca atagcatgat acaaaggcat taaagcagcg
     7681 tatccacata gcgtaaaagg agcaacatag ttaagaatac cagtcaatct ttcacaaatt
     7741 ttgtaatcca gaggttgatt atcgataagc ttggctgcag gtcgactcta gaaagtgtct
     7801 catgccggag gccattaaag acaggcctta ctggctgaat agaagcggtg ataattctta
     7861 attaattcat tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
     7921 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactgggt
     7981 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     8041 gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcgc
     8101 cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta
     8161 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     8221 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     8281 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     8341 gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt
     8401 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt
     8461 cagcttgccg taggtggcat cgccctcgcc ctcgccagac acgctgaact tgtggccgtt
     8521 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     8581 gctcaccatg gttgtggcca tattatcatc gtgtttttca aaggaaaacc acgtccccgt
     8641 ggttcggggg gcctagacgt ttttttaacc tcgactaaac acatgtaaag catgtgcacc
     8701 gaggccccag atcagatccc atacaatggg gtaccttctg ggcatccttc agccccttgt
     8761 tgaatacgct tgaggagagc catttgactc tttccacaac tatccaactc acaacgtggc
     8821 actggggttg tgccgccttt gcaggtgtat cttatacacg tggcttttgg ccgcagaggc
     8881 acctgtcgcc aggtgggggg ttccgctgcc tgcaaagggt cgctacagac gttgtttgtc
     8941 ttcaagaagc ttccagagga actgcttcct tcacgacatt caacagacct tgcattcctt
     9001 tggcgagagg ggaaagaccc ctaggaatgc tcgtcaagaa gacagggcca ggtttccggg
     9061 ccctcacatt gccaaaagac ggcaatatgg tggaaaataa catatagaca aacgcacacc
     9121 ggccttattc caagcggctt cggccagtaa cgttaggggg gggg
//
