LOCUS       sequence-329039-        8300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 3x Flag-tagged LGP2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8300)
  AUTHORS   Bamming D, Horvath CM
  TITLE     Regulation of signal transduction by enzymatically inactive 
            antiviral RNA helicase proteins MDA5, RIG-I, and LGP2.
  JOURNAL   J Biol Chem. 2009 Apr 10;284(15):9700-12. doi: 
            10.1074/jbc.M807365200. Epub 2009 Feb 11.
  PUBMED    19211564
REFERENCE   2  (bases 1 to 8300)
  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: 
            "10.1074/jbc.M807365200"; journalName: "J Biol Chem"; date: 
            "2009-04-10- 10"; volume: "284"; issue: "15"; pages: "9700-12"
COMMENT     Sequence Label: CMV-10-3xFLAG-LGP2
FEATURES             Location/Qualifiers
     source          1..8300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             614..679
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2742..3364
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2941..2960)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3388..3408)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3406..3703
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3757..4551
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3811..3830)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4421..4440
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5203..5337
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5240..5259)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5294..5313
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5342..5361)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5343..5361)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5375..5391)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5375..5391)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5388..5410)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5399..5415
                     /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(5423..5429,5430..5447,5448..5453))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5468..5489
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5606..5623)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5777..6365)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5857..5876)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6536..7327,7328..7396))
                     /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     7159..7178
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7397..7501)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7528..7983
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7615..7634)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7825..7846
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8109..8131
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8123..8140
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8124..8140
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattaacc atggactaca aagaccatga cggtgattat aaagatcatg acatcgatta
      661 caaggatgac gatgacaagc ttgcggccgc gatggagctt cggtcctacc aatgggaggt
      721 gatcatgcct gccctggagg gcaagaatat catcatctgg ctgcccacgg gtgccgggaa
      781 gacccgggcg gctgcttatg tggccaagcg gcacctagag actgtggatg gagccaaggt
      841 ggttgtattg gtcaacaggg tgcacctggt gacccagcat ggtgaagagt tcaggcgcat
      901 gctggatgga cgctggaccg tgacaaccct gagtggggac atgggaccac gtgctggctt
      961 tggccacctg gcccggtgcc atgacctgct catctgcaca gcagagcttc tgcagatggc
     1021 actgaccagc cccgaggagg aggagcacgt ggagctcact gtcttctccc tgatcgtggt
     1081 ggatgagtgc caccacacgc acaaggacac cgtctacaac gtcatcatga gccagtacct
     1141 agaacttaaa ctccagaggg cacagccgct accccaggtg ctgggtctca cagcctcccc
     1201 aggcactggc ggggcctcca aactcgatgg ggccatcaac cacgtcctgc agctctgtgc
     1261 caacttggac acgtggtgca tcatgtcacc ccagaactgc tgcccccagc tgcaggagca
     1321 cagccaacag ccttgcaaac agtacaacct ctgccacagg cgcagccagg atccgtttgg
     1381 ggacttgctg aagaagctca tggaccaaat ccatgaccac ctggagatgc ctgagttgag
     1441 ccggaaattt gggacgcaaa tgtatgagca gcaggtggtg aagctgagtg aggctgcggc
     1501 tttggctggg cttcaggagc aacgggtgta tgcgcttcac ctgaggcgct acaatgacgc
     1561 gctgctcatc catgacaccg tccgcgccgt ggatgccttg gctgcgctgc aggatttcta
     1621 tcacagggag cacgtcacta aaacccagat cctgtgtgcc gagcgccggc tgctggccct
     1681 gttcgatgac cgcaagaatg agctggccca cttggcaact catggcccag agaatccaaa
     1741 actggagatg ctggaaaaga tcctgcaaag gcagttcagt agctctaaca gccctcgggg
     1801 tatcatcttc acccgcaccc gccaaagcgc acactccctc ctgctctggc tccagcagca
     1861 gcagggcctg cagactgtgg acatccgggc ccagctactg attggggctg ggaacagcag
     1921 ccagagcacc cacatgaccc agagggacca gcaagaagtg atccagaagt tccaagatgg
     1981 aaccctgaac cttctggtgg ccacgagtgt ggcggaggag gggctggaca tcccacattg
     2041 caatgtggtg gtgcgttatg ggctcttgac caatgaaatc tccatggtcc aggccagggg
     2101 ccgtgcccgg gccgatcaga gtgtatacgc gtttgtagca actgaaggta gccgggagct
     2161 gaagcgggag ctgatcaacg aggcgctgga gacgctgatg gagcaggcag tggctgctgt
     2221 gcagaaaatg gaccaggccg agtaccaggc caagatccgg gatctgcagc aggcagcctt
     2281 gaccaagcgg gcggcccagg cagcccagcg ggagaaccag cggcagcagt tcccagtgga
     2341 gcacgtgcag ctactctgca tcaactgcat ggtggctgtg ggccatggca gcgacctgcg
     2401 gaaggtggag ggcacccacc atgtcaatgt gaaccccaac ttctcgaact actataatgt
     2461 ctccagggat cctgtggtca tcaacaaagt cttcaaggac tggaagcctg ggggtgtcat
     2521 cagctgcagg aactgtgggg aggtctgggg tctgcagatg atctacaagt cagtgaagct
     2581 gccagtgctc aaagtccgca gcatgctgct ggagacccct caggggcgga tccaggccaa
     2641 aaagtggtcc cgcgtgccct tctccgtgcc tgactttgac ttcctgcagc attgtgccga
     2701 gaacttgtcg gacctctccc tggactgatc tagaggatcc cgggtggcat ccctgtgacc
     2761 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     2821 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     2881 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     2941 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     3001 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     3061 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     3121 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     3181 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa taactatacc
     3241 agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg gtgggacatt
     3301 tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag atgcctgttg
     3361 aattgggtac gcggccagct tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3421 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3481 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3541 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3601 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3661 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggaatt gatcagcttg
     3721 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3781 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3841 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3901 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3961 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4021 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4081 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4141 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4201 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4261 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4321 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4381 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4441 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4501 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4561 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4621 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4681 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccgggctc gatcccctcg
     4741 cgagttggtt cagctgctgc ctgaggctgg acgacctcgc ggagttctac cggcagtgca
     4801 aatccgtcgg catccaggaa accagcagcg gctatccgcg catccatgcc cccgaactgc
     4861 aggagtgggg aggcacgatg gccgctttgg tcgacccgga cgggacgctc ctgcgcctga
     4921 tacagaacga attgcttgca ggcatctcat gagtgtgtct tcccgttttc cgcctgaggt
     4981 cactgcgtgg atggagcgct ggcgcctgct gcgcgacggc gagctgctca ccacccactc
     5041 gccaagctgg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgccgatc
     5101 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     5161 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     5221 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5281 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaat
     5341 tccctatagt gagtcgtatt aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt
     5401 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5461 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5521 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5581 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5641 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5701 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5761 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5821 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5881 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5941 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6001 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6061 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6121 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6181 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6241 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6301 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6361 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6421 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6481 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6541 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6601 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6661 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6721 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6781 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6841 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6901 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6961 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7021 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7081 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7141 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7201 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7261 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7321 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7381 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7441 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7501 gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg
     7561 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     7621 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     7681 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     7741 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     7801 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     7861 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     7921 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     7981 tttccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     8041 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     8101 agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagctga tctatacatt
     8161 gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa tcaatattgg
     8221 ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg
     8281 tccaatatga ccgccatgtt
//
