LOCUS       sequence-257011-        8345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MyD88 with an N-termial 3xFLAG tag and a C-terminal GyrB 
            domain from the E. coli DNA Gyrase B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8345)
  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 8345)
  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-GyrB
FEATURES             Location/Qualifiers
     source          1..8345
                     /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"
     CDS             1430..1495
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3149..3262
                     /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="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             3284..3328
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     misc_feature    3379..3945
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3395..3947
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3539..3556)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3766..3785
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3959..3961,3962..3964,3965..4678)
                     /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(4004..4025)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4265..4284)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4612..4633
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4740..5254
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5416..5432
                     /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(5440..5446,5447..5464,5465..5470))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5485..5506
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5623..5640)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5794..6382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5874..5893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6553..7344,7345..7413))
                     /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     7176..7195
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7414..7518)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7586..7604
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7642..7664)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7764..7783
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7977..7999
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8127..8146
                     /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 actacaaaga
     1441 ccatgatgga gactacaagg accatgacat cgactataag gacgacgacg ataagggagc
     1501 cggcgctggc gcgtctgccg gggaccctcg ggtgggaagt ggctccctcg actcattcat
     1561 gtttagtatt cccttggtgg ctctcaacgt gggggtgcgc cgacgcctct ccttgttttt
     1621 gaacccccga accccagttg ctgccgattg gaccctcctg gcagaggaga tggggttcga
     1681 gtacctcgag atacgagaat tggaaacccg acctgatcct acacgctcct tgctggacgc
     1741 ctggcaggga cgaagcggag ccagtgttgg gcgcttgctg gagctgctgg ctctgctgga
     1801 tcgagaagat atactcaagg agttgaagtc aaggattgag gaggactgcc aaaagtattt
     1861 gggtaaacag caaaaccaag aaagcgagaa accattgcaa gttgcccgcg ttgagtcttc
     1921 tgtgccccaa actaaggagc tcggtggcat caccaccctg gatgatcctt tgggccaaac
     1981 tccagaactg tttgacgcat ttatatgcta ctgccccaat gacattgagt tcgtgcagga
     2041 aatgattcga cagttggaac aaacagatta tcgactcaaa ttgtgtgtgt ctgaccgaga
     2101 tgttttgcca gggacttgtg tctggagcat tgcatctgaa cttattgaga agcgctgcag
     2161 gaggatggtg gtggtcgtgt cagacgatta ccttcagtcc aaagagtgtg atttccaaac
     2221 taagtttgcc cttagcctct ctccaggagt ccagcaaaag cggctcattc ccattaagta
     2281 taaagccatg aaaaaagatt tcccttctat tctccgattc atcaccattt gtgactacac
     2341 taatccttgt accaagagct ggttttggac taggctcgca aaggccctca gtttgcctgg
     2401 agccggagcc ggggcctcca attcatacga ttcttcctcc atcaaagtcc tgaaagggtt
     2461 ggatgctgtc cgcaagcgac caggcatgta tataggagat acagatgacg gcaccggcct
     2521 tcaccacatg gtattcgaag tggttgacaa tgccatagat gaggcacttg ccggacactg
     2581 caaggaaatc atagtaacca ttcatgctga taactcagtt tcagtccagg acgatggaag
     2641 gggtatccca actgggatac accccgaaga aggcgtttca gcagccgaag tgattatgac
     2701 cgtcctccat gctggaggaa agttcgacga taattcctac aaagtctccg gtggcctgca
     2761 tggggtgggc gtatctgtag taaatgctct cagccagaaa ctggaactcg tcattcagag
     2821 agaaggaaag atccaccgac agatatacga gcacggcgtc ccacaagctc ctctggcagt
     2881 tacaggggaa actgaaaaga caggcaccat ggtaaggttt tggccttccc ttgaaacttt
     2941 tacaaatgtg acagagttcg aatacgagat tctggccaaa cgattgaggg aactctcttt
     3001 tctcaactca ggagtgagta ttcggttgag ggataagcgg gacggaaaag aggaccactt
     3061 tcactacgaa ggaggtggtt cagaaggaaa atcttccgga tcaggaagcg aatcaaaggt
     3121 tacagactct ggagccggtg ctggagctat ggatgaaaag actacaggct ggcggggggg
     3181 acatgttgtg gaagggctgg caggggaact ggaacagttg cgcgcacgac ttgagcacca
     3241 ccctcagggc cagcgagaac cttctggggg ctgcaaactc ggcaaagaaa ccgcagctgc
     3301 aaagtttgag aggcagcaca tggacagctg agcggccgcg tttaaacgtc gacggtatcg
     3361 ataagcttga tatcgaattc cgcccccccc cctaacgtta ctggccgaag ccgcttggaa
     3421 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     3481 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     3541 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     3601 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     3661 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     3721 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     3781 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     3841 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     3901 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat
     3961 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     4021 cgacgtaaac ggccacaagt tcagcgtgtc tggcgagggc gagggcgatg ccacctacgg
     4081 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     4141 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     4201 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     4261 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     4321 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     4381 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     4441 catcaaggcg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     4501 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     4561 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     4621 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaatg
     4681 aattaattaa gaattcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     4741 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     4801 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
     4861 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     4921 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
     4981 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     5041 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     5101 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt gtatccaata
     5161 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
     5221 agtgattgac tacccgtcag cgggggtctt tcatgggtaa cagtttcttg aagttggaga
     5281 acaacattct gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt
     5341 tgaatccaca tactccaata ctcctgaaat agttcattat ggacagcgca gaaagagctg
     5401 gggagaattg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5461 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5521 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5581 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5641 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5701 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5761 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5821 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5881 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5941 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6001 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6061 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6121 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6181 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6241 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6301 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6361 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6421 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6481 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6541 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6601 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6661 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6721 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6781 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6841 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6901 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6961 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7021 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7081 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7141 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7201 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7261 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7321 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7381 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7441 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7501 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     7561 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     7621 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     7681 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     7741 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     7801 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     7861 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     7921 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     7981 gtcacgacgt tgtaaaacga cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac
     8041 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     8101 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     8161 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggcgatt agtccaattt
     8221 gttaaagaca ggatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg
     8281 aagcctatag agtacgagcc atagataaaa taaaagattt tatttagtct ccagaaaaag
     8341 ggggg
//
