LOCUS       sequence-309314-        5902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase under the control of 8 copies of the Aedes 
            aegypti Rel2 NF-kappa-B site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5902)
  AUTHORS   Russell TA, Ayaz A, Davidson AD, Fernandez-Sesma A, Maringer K
  TITLE     Imd pathway-specific immune assays reveal NF-kappaB stimulation by 
            viral RNA PAMPs in Aedes aegypti Aag2 cells.
  JOURNAL   PLoS Negl Trop Dis. 2021 Feb 16;15(2):e0008524. doi: 
            10.1371/journal.pntd.0008524.
  PUBMED    33591970
REFERENCE   2  (bases 1 to 5902)
  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"; journalName: "PLoS Negl 
            Trop Dis."; date: "2021-02-16"; volume: "15(2):e0008524. doi"; 
            pages: " 10.1371/journal.pntd.0008524"
COMMENT     Sequence Label: pKM54
FEATURES             Location/Qualifiers
     source          1..5902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        32..136
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(137..205,206..997)
                     /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     complement(355..374)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1168..1756
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1657..1676
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1910..1927
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1942..2082
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2008..2027)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     polyA_signal    2321..2455
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2549..2683
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             3018..4670
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(3084..3102)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          4801..4866
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4996..5016
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5441..5575
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5848..5866)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
       61 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
      121 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
      181 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
      241 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
      301 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      361 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      421 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      481 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      541 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      601 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
      661 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
      721 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
      781 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
      841 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
      901 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
      961 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     1021 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     1081 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1141 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1201 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1261 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1321 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1381 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1441 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1501 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1561 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1621 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1681 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1741 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1801 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1861 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1921 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     1981 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     2041 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     2101 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     2161 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     2221 caggatcagc catatcacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2281 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt aacttgttta ttgcagctta
     2341 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2401 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcagcca
     2461 tatcacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     2521 gaaacataaa atgaatgcaa ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2581 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2641 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccaagctt cccgggcaga
     2701 tctctcgagg tcgacccggg gatccacgct ttcggtgatt tacgcaccac gctttcggtg
     2761 atttacgcac cacgctttcg gtgatttacg caccacgctt tcggtgattt acgcacaatt
     2821 cacgctttcg gtgatttacg caccacgctt tcggtgattt acgcaccacg ctttcggtga
     2881 tttacgcacc acgctttcgg tgatttacgc acagcttaat tagatctaat taatataaag
     2941 catataaaca gcccagcttg gagcgagtga ccatgggagc tcgaattcca gcttggcatt
     3001 ccggtactgt tggtaaaatg gaagacgcca aaaacataaa gaaaggcccg gcgccattct
     3061 atcctctaga ggatggaacc gctggagagc aactgcataa ggctatgaag agatacgccc
     3121 tggttcctgg aacaattgct tttacagatg cacatatcga ggtgaacatc acgtacgcgg
     3181 aatacttcga aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa
     3241 atcacagaat cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg
     3301 cgttatttat cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc
     3361 tcaacagtat gaacatttcg cagcctaccg tagtgtttgt ttccaaaaag gggttgcaaa
     3421 aaattttgaa cgtgcaaaaa aaattaccaa taatccagaa aattattatc atggattcta
     3481 aaacggatta ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg
     3541 gttttaatga atacgatttt gtaccagagt cctttgatcg tgacaaaaca attgcactga
     3601 taatgaattc ctctggatct actgggttac ctaagggtgt ggcccttccg catagaactg
     3661 cctgcgtcag attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata
     3721 ctgcgatttt aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat
     3781 atttgatatg tggatttcga gtcgtcttaa tgtatagatt tgaagaagag ctgtttttac
     3841 gatcccttca ggattacaaa attcaaagtg cgttgctagt accaacccta ttttcattct
     3901 tcgccaaaag cactctgatt gacaaatacg atttatctaa tttacacgaa attgcttctg
     3961 ggggcgcacc tctttcgaaa gaagtcgggg aagcggttgc aaaacgcttc catcttccag
     4021 ggatacgaca aggatatggg ctcactgaga ctacatcagc tattctgatt acacccgagg
     4081 gggatgataa accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg
     4141 atctggatac cgggaaaacg ctgggcgtta atcagagagg cgaattatgt gtcagaggac
     4201 ctatgattat gtccggttat gtaaacaatc cggaagcgac caacgccttg attgacaagg
     4261 atggatggct acattctgga gacatagctt actgggacga agacgaacac ttcttcatag
     4321 ttgaccgctt gaagtcttta attaaataca aaggatatca ggtggccccc gctgaattgg
     4381 aatcgatatt gttacaacac cccaacatct tcgacgcggg cgtggcaggt cttcccgacg
     4441 atgacgccgg tgaacttccc gccgccgttg ttgttttgga gcacggaaag acgatgacgg
     4501 aaaaagagat cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag
     4561 gagttgtgtt tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa
     4621 tcagagagat cctcataaag gccaagaagg gcggaaagtc caaattgtaa aatgtaactg
     4681 tattcagcga tgacgaaatt cttagctatt gtaatactct agaggatctt tgtgaaggaa
     4741 ccttacttct gtggtgtgac ataattggac aaactaccta cagagattta aagctctaag
     4801 gtaaatataa aatttttaag tgtataatgt gttaaactac tgattctaat tgtttgtgta
     4861 ttttagattc caacctatgg aactgatgaa tgggagcagt ggtggaatgc ctttaatgag
     4921 gaaaacctgt tttgctcaga agaaatgcca tctagtgatg atgaggctac tgctgactct
     4981 caacattcta ctcctccaaa aaagaagaga aaggtagaag accccaagga ctttccttca
     5041 gaattgctaa gttttttgag tcatgctgtg tttagtaata gaactcttgc ttgctttgct
     5101 atttacacca caaaggaaaa agctgcactg ctatacaaga aaattatgga aaaatattct
     5161 gtaaccttta taagtaggca taacagttat aatcataaca tactgttttt tcttactcca
     5221 cacaggcata gagtgtctgc tattaataac tatgctcaaa aattgtgtac ctttagcttt
     5281 ttaatttgta aaggggttaa taaggaatat ttgatgtata gtgccttgac tagagatcat
     5341 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     5401 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     5461 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5521 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccccag
     5581 gaagctcctc tgtgtcctca taaaccctaa cctcctctac ttgagaggac attccaatca
     5641 taggctgccc atccaccctc tgtgtcctcc tgttaattag gtcacttaac aaaaaggaaa
     5701 ttgggtaggg gtttttcaca gaccgctttc taagggtaat tttaaaatat ctgggaagtc
     5761 ccttccactg ctgtgttcca gaagtgttgg taaacagccc acaaatgtca acagcagaaa
     5821 catacaagct gtcagctttg cacaagggcc tcgtgatacg cctattttta taggttaatg
     5881 tcatgataat aatggtttct ta
//
