LOCUS       sequence-204696-        7775 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7775)
  AUTHORS   Quarato G, Guy CS, Grace CR, Llambi F, Nourse A, Rodriguez DA, 
            Wakefield R, Frase S, Moldoveanu T, Green DR
  TITLE     Sequential Engagement of Distinct MLKL Phosphatidylinositol-Binding 
            Sites Executes Necroptosis.
  JOURNAL   Mol Cell. 2016 Feb 18;61(4):589-601. doi: 
            10.1016/j.molcel.2016.01.011. Epub 2016 Feb 4.
  PUBMED    26853145
REFERENCE   2  (bases 1 to 7775)
  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.molcel.2016.01.011"; journalName: "Mol Cell"; date: 
            "2016-02-18- 18"; volume: "61"; issue: "4"; pages: "589-601"
COMMENT     Sequence Label: Cerulean-2xFv-hRIPK3
FEATURES             Location/Qualifiers
     source          1..7775
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    580..937
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1002..1418
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1348..1370
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1386..1402
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1428..1802
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1740..1756
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(1814..1816,1817..1819,1820..2530)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1859..1880)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2120..2139)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2467..2488
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2537..2857
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2864..3184
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     LTR             4880..5349
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     rep_origin      complement(5892..6480)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5972..5991)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6644..7435,7436..7504))
                     /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     7267..7286
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7505..7609)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag
      121 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg wmcccagmtg mrgtccagcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
      421 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
      481 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
      541 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggccag caacttatct
      601 gtgtctgtcc gattgtctag tgtctatgac tgattttatg cgcctgcgtc ggtactagtt
      661 agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttcg gaacacccgg
      721 ccgcaaccct gggagacgtc ccagggactt cgggggccgt ttttgtggcc cgacctgagt
      781 ccaaaaatcc cgatcgtttt ggactctttg gtgcaccccc cttagaggag ggatatgtgg
      841 ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg
      901 tttgggaccg aagccgcgcc gcgcgtcttg tctgctgcag catcgttctg tgttgtctct
      961 gtctgactgt gtttctgtat ttgtctgaaa atatgggccc gggccagact gttaccactc
     1021 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1081 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1141 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1201 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1261 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1321 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1381 cctcccttta tccagccctc actccttctc taggcgcccc catatggcca tatgagatct
     1441 tatatggggc acccccgccc cttgtaaact tccctgaccc tgacatgaca agagttacta
     1501 acagcccctc tctccaagct cacttacagg ctctctactt agtccagcac gaagtctgga
     1561 gacctctggc ggcagcctac caagaacaac tggaccgacc ggtggtacct cacccttacc
     1621 gagtcggcga cacagtgtgg gtccgccgac accagactaa gaacctagaa cctcgctgga
     1681 aaggacctta cacagtcctg ctgaccaccc ccaccgccct caaagtagac ggcatcgcag
     1741 cttggataca cgccgcccac gtgaaggctg ccgaccccgg gggtggacca tcctctagac
     1801 tgccggatct accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1861 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1921 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1981 gccctggccc accctcgtga ccaccctgac ctggggcgtg cagtgcttcg cccgctaccc
     2041 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2101 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2161 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2221 catcctgggg cacaagctgg agtacaacgc catcagcgac aacgtctata tcaccgccga
     2281 caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcag
     2341 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2401 gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg
     2461 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2521 gctgtacaag tctagaggcg tccaagtcga aaccattagt cccggcgatg gcagaacatt
     2581 tcctaaaagg ggacaaacat gtgtcgtcca ttatacaggc atgttggagg acggcaaaaa
     2641 ggtggacagt agtagagatc gcaataaacc tttcaaattc atgttgggaa aacaagaagt
     2701 cattagggga tgggaggagg gcgtggctca aatgtccgtc ggccaacgcg ctaagctcac
     2761 catcagcccc gactacgcat acggcgctac cggacatccc ggaattattc cccctcacgc
     2821 taccttggtg tttgacgtcg aactgttgaa gctcgagact agaggagtgc aggtggagac
     2881 tatctcccca ggagacgggc gcaccttccc caagcgcggc cagacctgcg tggtgcacta
     2941 caccgggatg cttgaagatg gaaagaaagt tgattcctcc cgggacagaa acaagccctt
     3001 taagtttatg ctaggcaagc aggaggtgat ccgaggctgg gaagaagggg ttgcccagat
     3061 gagtgtgggt cagagagcca aactgactat atctccagat tatgcctatg gtgccactgg
     3121 gcacccaggc atcatcccac cacatgccac tctcgtcttc gatgtggagc ttctaaaact
     3181 ggaaactagt atgtcgtgcg tcaagttatg gcccagcggt gcccccgccc ccttggtgtc
     3241 catcgaggaa ctggagaacc aggagctcgt cggcaaaggc gggttcggca cagtgttccg
     3301 ggcgcaacat aggaagtggg gctacgatgt ggcggtcaag atcgtaaact cgaaggcgat
     3361 atccagggag gtcaaggcca tggcaagtct ggataacgaa ttcgtgctgc gcctagaagg
     3421 ggttatcgag aaggtgaact gggaccaaga tcccaagccg gctctggtga ctaaattcat
     3481 ggagaacggc tccttgtcgg ggctgctgca gtcccagtgc cctcggccct ggccgctcct
     3541 ttgccgcctg ctgaaagaag tggtgcttgg gatgttttac ctgcacgacc agaacccggt
     3601 gctcctgcac cgggacctca agccatccaa cgtcctgctg gacccagagc tgcacgtcaa
     3661 gctggcagat tttggcctgt ccacatttca gggaggctca cagtcaggga cagggtccgg
     3721 ggagccaggg ggcaccctgg gctacttggc cccagaactg tttgttaacg taaaccggaa
     3781 ggcctccaca gccagtgacg tctacagctt cgggatccta atgtgggcag tgcttgctgg
     3841 aagagaagtt gagttgccaa ccgaaccatc actcgtgtac gaagcagtgt gcaacaggca
     3901 gaaccggcct tcattggctg agctgcccca agccgggcct gagactcccg gcttagaagg
     3961 actgaaggag ctaatgcagc tctgctggag cagtgagccc aaggacagac cctccttcca
     4021 ggaatgccta ccaaaaactg atgaagtctt ccagatggtg gagaacaata tgaatgctgc
     4081 tgtctccacg gtaaaggatt tcctgtctca gctcaggagc agcaatagga gattttctat
     4141 cccagagtca ggccaaggag ggacagaaat ggatggcttt aggagaacca tagaaaacca
     4201 gcactctcgt aatgatgtca tggtttctga gtggctaaac aaactgaatc tagaggagcc
     4261 tcccagctct gttcctaaaa aatgcccgag ccttaccaag aggagcaggg cacaagagga
     4321 gcaggttcca caagcctgga cagcaggcac atcttcagat tcgatggccc aacctcccca
     4381 gactccagag acctcaactt tcagaaacca gatgcccagc cctacctcaa ctggaacacc
     4441 aagtcctgga ccccgaggga atcagggggc tgagagacaa ggcatgaact ggtcctgcag
     4501 gaccccggag ccaaatccag taacagggcg accgctcgtt aacatataca actgctctgg
     4561 ggtgcaagtt ggagacaaca actacttgac tatgcaacag acaactgcct tgcccacatg
     4621 gggcttggca ccttcgggca aggggagggg cttgcagcac cccccaccag taggttcgca
     4681 agaaggccct aaagatcctg aagcctggag caggccacag ggttggtata atcatagcgg
     4741 gaaataagtc gacgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     4801 aagaccccac ctgtaggttt ggcaagctag cttaagtaam gccatttkky cagatcaagg
     4861 tcaggaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     4921 tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct
     4981 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     5041 agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     5101 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     5161 tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg
     5221 attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac
     5281 ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg
     5341 ggtctttcac atgcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat
     5401 taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5461 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5521 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5581 aggaaagwrm rytaactcac attaattgcg ttgckytcrc tkyyykcytt mmrkwykkrm
     5641 awymwrtsgc catagttgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5701 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5761 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5821 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5881 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5941 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6001 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6061 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6121 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6181 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6241 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6301 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6361 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6421 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6481 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6541 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt gcggccggcc
     6601 gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6661 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6721 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6781 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6841 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6901 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6961 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7021 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7081 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7141 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7201 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7261 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7321 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7381 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7441 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7501 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7561 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7621 gaaaagtgcc acctgcataa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt
     7681 aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagaaaagt tcagatcaag
     7741 gtcaggaaca gatggaacag ctgaatatgg gccaa
//
