LOCUS Exported 7030 bp ds-DNA circular SYN 12-MAY-2021 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 mscv2.2-6mycPrgJ 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7030 /organism="synthetic DNA construct" /mol_type="other DNA" 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 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 1345..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(2848..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(3961..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 6742..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 //