LOCUS       sequence-96380-e        8151 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  6xMYC-tagged full-length FlaA from Legionella pneumophila, expressed
            under the constitutive, moderate-level retroviral LTR promoter for 
            mammalian cell expression; IRES-GFP reporter downstream of FlaA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8151)
  AUTHORS   Kofoed EM, Vance RE
  TITLE     Innate immune recognition of bacterial ligands by NAIPs determines 
            inflammasome specificity.
  JOURNAL   Nature. 2011 Aug 28;477(7366):592-5. doi: 10.1038/nature10394.
  PUBMED    21874021
REFERENCE   2  (bases 1 to 8151)
  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"; journalName: 
            "Nature"; date: "2011-08-28- 28"; volume: "477"; issue: "7366"; 
            pages: "592-5"
COMMENT     Sequence Label: mscv2.2-6mycFlaA
FEATURES             Location/Qualifiers
     source          1..8151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..80
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     96..112
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             158..187
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             197..226
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             236..265
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             275..304
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             314..343
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             377..406
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1886..2452
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1902..2454
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2046..2063)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2273..2292
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2466..2468,2469..2471,2472..3185)
                     /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(2511..2532)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2772..2791)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3119..3140
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3238..3752
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(3921..3937)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3921..3937)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3934..3956)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3945..3961
                     /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(3969..3975,3976..3993,3994..3999))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4014..4035
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4152..4169)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4323..4911)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4403..4422)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5082..5873,5874..5942))
                     /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     5705..5724
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5943..6047)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6115..6133
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6171..6193)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6293..6312
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6506..6528
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6656..6675
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6875..7391
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7455..7796
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(7863..8151,1..128)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
       61 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
      121 taggcgccgg aattagatct atcgatttaa agctatggag caaaagctca tttctgaaga
      181 ggacttgaat gaaatggagc aaaagctcat ttctgaagag gacttgaatg aaatggagca
      241 aaagctcatt tctgaagagg acttgaatga aatggagcaa aagctcattt ctgaagagga
      301 cttgaatgaa atggagcaaa agctcatttc tgaagaggac ttgaatgaaa tggagagctt
      361 gggcgacctc accatggagc aaaagctcat ttctgaagag gacttgaatt caggccggcc
      421 agcggccgca gctcaagtaa tcaacactaa tgtggcgtcg ctcacagccc aacgtaattt
      481 gggtgtttcg ggcaacatga tgcaaacatc gatccagcgt ttatcatcgg gattaaggat
      541 taacagtgcc aaagatgatg cagcaggact cgccatttct caacgaatga ctgcacaaat
      601 tcgcgggatg aaccaagccg ttagaaatgc caacgatggt atttccctgg ctcaggttgc
      661 tgaaggagca atgcaggaaa caaccaatat tttacaacgc atgcgtgagc tttccgttca
      721 agctgctaac tcaaccaata actcatctga ccggtcttca attcaaagtg aaatttcaca
      781 attaaaaagt gaattggagc gtattgctca aaatactgaa ttcaatggtc aaagaattct
      841 cgatggttct ttctctggcg caagcttcca ggttggggcg aattcaaacc aaaccatcaa
      901 tttcagtatc ggcagcacaa aagcttcttc tcttggtggt attgccacgg caacaggaac
      961 agaagtagca ggtgcagcag cggcagatat tactatcgca attggaggag gagcagcaac
     1021 cagtattaac tcttctgcca attttaccgg ggcactaaac ggacaagatg ctacgtcagc
     1081 ctatgccaaa gcagcggcta tcaacgatgc cgggatagga ggactatcgg ttacagcgtc
     1141 taccagcggc acacaagcag ttggcgcaat aggtggaact gcaggtgata cctacaactt
     1201 aacgattaat ggtgttgcaa tatacactaa ccttaacgta gcaacggcat taaccaactc
     1261 tgatcttaga gatgccatca atggcgtcag taaccaaact ggagttgttg catcacttaa
     1321 cggtggtaat atgacactca ctgctgctga tggcagaaac attacggtaa cagaaagcgg
     1381 tacaggtttt actgcaggta ctgacggttt aactgtaact ggtggcgcat ttgatggcgc
     1441 gctacgcggt actttatcaa tcagcgccgt tgatactatc gcgattggtg gtactgttgc
     1501 taacattggt ttaagtgcca atatctctaa agacacagta gggatagatt cactggatgt
     1561 gagcacagcg tctggcgctc aaacagcgat taagagaatt gatgcagcac tcaatagcgt
     1621 aaactctaat cgggctaata tgggtgctct gcaaaatcgc tttgaatcaa cgatagccaa
     1681 cctgcaaaac gtatcagaca acttatcagc tgctagaagt cgtattcaag atgctgatta
     1741 cgctgctgaa atggctagtt tgaccaaaaa ccaaattctg caacaagcag gtacagcgat
     1801 gttggcacaa gctaatagcc taccgcaatc tgtattatct ttgttaggtc gatagtcgac
     1861 ggtatcgata agcttgatat cgaattccgc ccccccccct aacgttactg gccgaagccg
     1921 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
     1981 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
     2041 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
     2101 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
     2161 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
     2221 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
     2281 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
     2341 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
     2401 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca
     2461 caaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2521 tggacggcga cgtaaacggc cacaagttca gcgtgtctgg cgagggcgag ggcgatgcca
     2581 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     2641 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     2701 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     2761 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2821 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     2881 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     2941 agaacggcat caaggcgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     3001 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3061 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     3121 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3181 agtgataaga attcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg
     3241 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg
     3301 aaaatacata actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga
     3361 atatgggcca aacaggatat ctgtggtaag cagttcctgc ccccggctca gggccaagaa
     3421 cagatggtcc ccgatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc
     3481 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     3541 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     3601 cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgtgt atccaataaa
     3661 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     3721 tgattgacta cccgtcagcg ggggtctttc atgggtaaca gtttcttgaa gttggagaac
     3781 aacattctga gggtaggagt cgaatattaa gtaatcctga ctcaattagc cactgttttg
     3841 aatccacata ctccaatact cctgaaatag ttcattatgg acagcgcaga agagctgggg
     3901 agaattaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3961 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4021 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4081 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4141 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4201 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4261 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4321 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4381 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4441 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4501 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4561 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4621 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4681 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4741 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     4801 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4861 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4921 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4981 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5041 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5101 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5161 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5221 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5281 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5341 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5401 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5461 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5521 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5581 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5641 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5701 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5761 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5821 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5881 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5941 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6001 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6061 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     6121 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     6181 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     6241 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     6301 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     6361 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     6421 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     6481 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggcgcaagga
     6541 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     6601 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     6661 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     6721 gtccggcgta gaggcgatta gtccaatttg ttaaagacag gatatcagtg gtccaggctc
     6781 tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca tagataaaat
     6841 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     6901 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga
     6961 gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct
     7021 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     7081 cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     7141 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     7201 tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg
     7261 atagactgcg tcgcccgggt acccgtattc ccaataaagc ctcttgctgt ttgcatccga
     7321 atcgtggact cgctgatcct tgggagggtc tcctcagatt gattgactgc ccacctcggg
     7381 ggtctttcat ttggaggttc caccgagatt tggagacccc tgcccaggga ccaccgaccc
     7441 ccccgccggg aggtaagctg gccagcggtc gtttcgtgtc tgtctctgtc tttgtgcgtg
     7501 tttgtgccgg catctaatgt ttgcgcctgc gtctgtacta gttagctaac tagctctgta
     7561 tctggcggac ccgtggtgga actgacgagt tctgaacacc cggccgcaac cctgggagac
     7621 gtcccaggga ctttgggggc cgtttttgtg gcccgacctg aggaagggag tcgatgtgga
     7681 atccgacccc gtcaggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc
     7741 ctccgtctga atttttgctt tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc
     7801 gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt
     7861 agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc
     7921 ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg
     7981 cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca
     8041 tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc
     8101 cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc a
//
