LOCUS       sequence-96381-e        7030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  6myc-tagged PrgJ (SPI-1 rod protein) from Salmonella typhimurium, 
            under the constitutive, moderate-level retroviral LTR promoter for 
            mammalian cell expression; IRES-GFP reporter downstream of PrgJ.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7030)
  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 7030)
  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: mscv2.2-6mycPrgJ
FEATURES             Location/Qualifiers
     source          1..7030
                     /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    765..1331
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    781..1333
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(925..942)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1152..1171
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1345..1347,1348..1350,1351..2064)
                     /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(1390..1411)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1651..1670)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1998..2019
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2117..2631
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(2800..2816)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2800..2816)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2813..2835)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2824..2840
                     /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(2848..2854,2855..2872,2873..2878))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2893..2914
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3031..3048)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3202..3790)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3282..3301)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3961..4752,4753..4821))
                     /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     4584..4603
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4822..4926)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4994..5012
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5050..5072)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5172..5191
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5385..5407
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5535..5554
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5754..6270
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6334..6675
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(6742..7030,1..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 tcgattgcaa ctattgtccc tgagaatgcc gttatagggc aggcggtcaa
      481 tatcaggtct atggaaacgg acattgtctc gctggatgac cggctactcc aggctttttc
      541 tggttcggcg attgccacgg ctgtggataa acagacgatt accaacagga ttgaggaccc
      601 taatctggtg acggatccta aagagctggc tatttcgcaa gagatgattt cagattataa
      661 cctgtatgtt tctatggtca gtacccttac tcgtaaagga gtcggggctg ttgaaacgct
      721 attacgctca tgagtcgacg gtatcgataa gcttgatatc gaattccgcc ccccccccta
      781 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
      841 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
      901 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
      961 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     1021 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     1081 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     1141 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     1201 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     1261 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     1321 cacgatgata atatggccac aaccatggtg agcaagggcg aggagctgtt caccggggtg
     1381 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtctggc
     1441 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     1501 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     1561 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1621 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1681 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1741 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1801 atcatggccg acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacatc
     1861 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1921 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     1981 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     2041 ggcatggacg agctgtacaa gtgataagaa ttcgataaaa taaaagattt tatttagtct
     2101 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac
     2161 gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta
     2221 ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     2281 cccggctcag ggccaagaac agatggtccc cgatgcggtc ccgccctcag cagtttctag
     2341 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     2401 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     2461 aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc gtcgcccggg
     2521 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     2581 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tgggtaacag
     2641 tttcttgaag ttggagaaca acattctgag ggtaggagtc gaatattaag taatcctgac
     2701 tcaattagcc actgttttga atccacatac tccaatactc ctgaaatagt tcattatgga
     2761 cagcgcagaa gagctgggga gaattaattc gtaatcatgg tcatagctgt ttcctgtgtg
     2821 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     2881 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2941 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3001 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3061 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3121 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3181 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3241 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3301 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3361 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3421 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3481 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3541 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3601 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     3661 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3721 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3781 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3841 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3901 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3961 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4021 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4081 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4141 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4201 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4261 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4321 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4381 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4441 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4501 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4561 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4621 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4681 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4741 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4801 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4861 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4921 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4981 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     5041 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     5101 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     5161 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     5221 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     5281 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     5341 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     5401 taaaacgacg gcgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     5461 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     5521 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     5581 gtgatgccgg ccacgatgcg tccggcgtag aggcgattag tccaatttgt taaagacagg
     5641 atatcagtgg tccaggctct agttttgact caacaatatc accagctgaa gcctatagag
     5701 tacgagccat agataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa
     5761 gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa
     5821 atacataact gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata
     5881 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     5941 tggtccccag atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg
     6001 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
     6061 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
     6121 tcggcgcgcc agtcctccga tagactgcgt cgcccgggta cccgtattcc caataaagcc
     6181 tcttgctgtt tgcatccgaa tcgtggactc gctgatcctt gggagggtct cctcagattg
     6241 attgactgcc cacctcgggg gtctttcatt tggaggttcc accgagattt ggagacccct
     6301 gcccagggac caccgacccc cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct
     6361 gtctctgtct ttgtgcgtgt ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag
     6421 ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc
     6481 ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga
     6541 ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag
     6601 aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc
     6661 gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt
     6721 ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt
     6781 cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt
     6841 tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc
     6901 acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat
     6961 ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct
     7021 ccctgggtca
//
