LOCUS       sequence-263594-        7709 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MyD88 tagged with a 3xFLAG tag at its N-terminus and a V5 
            tag at its C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7709)
  AUTHORS   Tan Y, Kagan JC
  TITLE     Innate Immune Signaling Organelles Display Natural and Programmable 
            Signaling Flexibility.
  JOURNAL   Cell. 2019 Apr 4;177(2):384-398.e11. doi: 
            10.1016/j.cell.2019.01.039. Epub 2019 Mar 7.
  PUBMED    30853218
REFERENCE   2  (bases 1 to 7709)
  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.2019.01.039"; journalName: "Cell"; date: "2019-04-4-
            4"; volume: "177"; issue: "2"; pages: "384-398.e11"
COMMENT     Sequence Label: pMSCV-IRES-GFP-3xFLAG-MyD88-V5
FEATURES             Location/Qualifiers
     source          1..7709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1421..1430
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1457..1466
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1466..1531
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2456..2497
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2516..2626
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             2648..2692
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     misc_feature    2743..3309
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2759..3311
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2903..2920)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3130..3149
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3323..3325,3326..3328,3329..4042)
                     /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(3368..3389)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3629..3648)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3976..3997
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4104..4618
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4780..4796
                     /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(4804..4810,4811..4828,4829..4834))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4849..4870
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4987..5004)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5158..5746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5238..5257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5917..6708,6709..6777))
                     /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     6540..6559
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6778..6882)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6950..6968
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7006..7028)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7128..7147
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7341..7363
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7491..7510
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tagatctgcc gccaccatgg attacaagga
     1441 cataataaga tctgccgcca ccatggatta caaggaccat gatggcgact ataaggacca
     1501 tgatatcgat tataaagacg acgacgacaa gggtgctggg gcgggtgcga tgagtgcagg
     1561 ggatccacgc gttgggagtg gctcattgga ttcattcatg ttctccattc ccttagtagc
     1621 tctgaacgtt ggagtgcgtc gtcgcctgtc cttgttcctt aatccccgca cgccggtcgc
     1681 agcagattgg actttattgg cggaggaaat gggcttcgag taccttgaaa ttcgtgaact
     1741 tgagacccgc ccagacccaa ctcgtagctt attggacgct tggcaggggc gtagtggcgc
     1801 atctgtcggg cgtcttcttg agctgctggc cctgttggac cgtgaggaca ttttgaaaga
     1861 gctgaaatcc cgtatcgaag aggactgtca gaaatacctt ggaaaacagc agaaccaaga
     1921 gtccgagaag cccttacagg tagcccgcgt cgagtcatca gttcctcaga caaaagaatt
     1981 ggggggcatc accacattag acgatccttt gggtcaaaca cctgagttat ttgatgcctt
     2041 tatttgttat tgccctaatg atattgaatt cgttcaggaa atgatccgtc aacttgagca
     2101 gacggactat cgtctgaaat tgtgtgtctc cgaccgtgac gtattaccgg ggacttgtgt
     2161 atggtcaatt gccagcgaat tgatcgagaa acgttgtcgc cgcatggtcg tcgttgtgtc
     2221 ggatgattac ctgcaatcta aagaatgtga cttccagacc aagtttgctt taagtttgtc
     2281 tccaggggtt caacagaaac gtttgatccc catcaaatat aaagcaatga agaaagactt
     2341 tccctcaatt ttgcgcttta tcacgatctg cgattacaca aacccatgta ccaaatcttg
     2401 gttttggacc cgtctggcaa aggcactgtc cttaccgggc gccggggcgg gagcaggtaa
     2461 accaattccc aatcctcttt tgggccttga ttcaacgggt gcgggtgcgg gcgccgacga
     2521 gaaaaccacc gggtggcgcg ggggtcacgt cgtagaaggg ttagcggggg aattagaaca
     2581 gcttcgtgct cgcttagaac accaccccca aggccagcgc gagccttcag gcgggtgcaa
     2641 acttggcaag gaaaccgcgg cagcgaaatt tgagcgtcag catatggatt catgagcggc
     2701 cgcgtttaaa cgtcgacggt atcgataagc ttgatatcga attccgcccc cccccctaac
     2761 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     2821 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     2881 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     2941 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     3001 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     3061 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     3121 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     3181 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     3241 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     3301 cgatgataat atggccacaa ccatggtgag caagggcgag gagctgttca ccggggtggt
     3361 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtctggcga
     3421 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3481 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     3541 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3601 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     3661 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3721 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     3781 catggccgac aagcagaaga acggcatcaa ggcgaacttc aagatccgcc acaacatcga
     3841 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3901 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     3961 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     4021 catggacgag ctgtacaagt aatgaattaa ttaagaattc gataaaataa aagattttat
     4081 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt
     4141 aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc
     4201 aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt
     4261 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag
     4321 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
     4381 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
     4441 gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc
     4501 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
     4561 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcatgg
     4621 gtaacagttt cttgaagttg gagaacaaca ttctgagggt aggagtcgaa tattaagtaa
     4681 tcctgactca attagccact gttttgaatc cacatactcc aatactcctg aaatagttca
     4741 ttatggacag cgcagaaaga gctggggaga attgtgaaat tgttatccgc tcacaattcc
     4801 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4861 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4921 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4981 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5041 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5101 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5161 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5221 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5281 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5341 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5401 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5461 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5521 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5581 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5641 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5701 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5761 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5821 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5881 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5941 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6001 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6061 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6121 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6181 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6241 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6301 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6361 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6421 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6481 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6541 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6601 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6661 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6721 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6781 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6841 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6901 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     6961 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     7021 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     7081 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     7141 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     7201 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     7261 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     7321 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt
     7381 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     7441 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     7501 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     7561 gcgtagaggc gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt
     7621 ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag
     7681 attttattta gtctccagaa aaagggggg
//
