LOCUS       sequence-309130-        5463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase with no promoter for negative control for 
            luciferase assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5463)
  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 5463)
  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: pKM45
FEATURES             Location/Qualifiers
     source          1..5463
                     /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"
     CDS             2579..4231
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2645..2663)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          4362..4427
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4557..4577
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5002..5136
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5409..5427)
                     /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 ggatccaagc
     2461 ttcccgggca gatctctcga ggtcgacaat acaatataaa gcatataaac agcccagctt
     2521 ggagcgagtg accatgggag ctcgaattcc agcttggcat tccggtactg ttggtaaaat
     2581 ggaagacgcc aaaaacataa agaaaggccc ggcgccattc tatcctctag aggatggaac
     2641 cgctggagag caactgcata aggctatgaa gagatacgcc ctggttcctg gaacaattgc
     2701 ttttacagat gcacatatcg aggtgaacat cacgtacgcg gaatacttcg aaatgtccgt
     2761 tcggttggca gaagctatga aacgatatgg gctgaataca aatcacagaa tcgtcgtatg
     2821 cagtgaaaac tctcttcaat tctttatgcc ggtgttgggc gcgttattta tcggagttgc
     2881 agttgcgccc gcgaacgaca tttataatga acgtgaattg ctcaacagta tgaacatttc
     2941 gcagcctacc gtagtgtttg tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa
     3001 aaaattacca ataatccaga aaattattat catggattct aaaacggatt accagggatt
     3061 tcagtcgatg tacacgttcg tcacatctca tctacctccc ggttttaatg aatacgattt
     3121 tgtaccagag tcctttgatc gtgacaaaac aattgcactg ataatgaatt cctctggatc
     3181 tactgggtta cctaagggtg tggcccttcc gcatagaact gcctgcgtca gattctcgca
     3241 tgccagagat cctatttttg gcaatcaaat cattccggat actgcgattt taagtgttgt
     3301 tccattccat cacggttttg gaatgtttac tacactcgga tatttgatat gtggatttcg
     3361 agtcgtctta atgtatagat ttgaagaaga gctgttttta cgatcccttc aggattacaa
     3421 aattcaaagt gcgttgctag taccaaccct attttcattc ttcgccaaaa gcactctgat
     3481 tgacaaatac gatttatcta atttacacga aattgcttct gggggcgcac ctctttcgaa
     3541 agaagtcggg gaagcggttg caaaacgctt ccatcttcca gggatacgac aaggatatgg
     3601 gctcactgag actacatcag ctattctgat tacacccgag ggggatgata aaccgggcgc
     3661 ggtcggtaaa gttgttccat tttttgaagc gaaggttgtg gatctggata ccgggaaaac
     3721 gctgggcgtt aatcagagag gcgaattatg tgtcagagga cctatgatta tgtccggtta
     3781 tgtaaacaat ccggaagcga ccaacgcctt gattgacaag gatggatggc tacattctgg
     3841 agacatagct tactgggacg aagacgaaca cttcttcata gttgaccgct tgaagtcttt
     3901 aattaaatac aaaggatatc aggtggcccc cgctgaattg gaatcgatat tgttacaaca
     3961 ccccaacatc ttcgacgcgg gcgtggcagg tcttcccgac gatgacgccg gtgaacttcc
     4021 cgccgccgtt gttgttttgg agcacggaaa gacgatgacg gaaaaagaga tcgtggatta
     4081 cgtcgccagt caagtaacaa ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga
     4141 agtaccgaaa ggtcttaccg gaaaactcga cgcaagaaaa atcagagaga tcctcataaa
     4201 ggccaagaag ggcggaaagt ccaaattgta aaatgtaact gtattcagcg atgacgaaat
     4261 tcttagctat tgtaatactc tagaggatct ttgtgaagga accttacttc tgtggtgtga
     4321 cataattgga caaactacct acagagattt aaagctctaa ggtaaatata aaatttttaa
     4381 gtgtataatg tgttaaacta ctgattctaa ttgtttgtgt attttagatt ccaacctatg
     4441 gaactgatga atgggagcag tggtggaatg cctttaatga ggaaaacctg ttttgctcag
     4501 aagaaatgcc atctagtgat gatgaggcta ctgctgactc tcaacattct actcctccaa
     4561 aaaagaagag aaaggtagaa gaccccaagg actttccttc agaattgcta agttttttga
     4621 gtcatgctgt gtttagtaat agaactcttg cttgctttgc tatttacacc acaaaggaaa
     4681 aagctgcact gctatacaag aaaattatgg aaaaatattc tgtaaccttt ataagtaggc
     4741 ataacagtta taatcataac atactgtttt ttcttactcc acacaggcat agagtgtctg
     4801 ctattaataa ctatgctcaa aaattgtgta cctttagctt tttaatttgt aaaggggtta
     4861 ataaggaata tttgatgtat agtgccttga ctagagatca taatcagcca taccacattt
     4921 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     4981 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     5041 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5101 tccaaactca tcaatgtatc ttatcatgtc tggatcccca ggaagctcct ctgtgtcctc
     5161 ataaacccta acctcctcta cttgagagga cattccaatc ataggctgcc catccaccct
     5221 ctgtgtcctc ctgttaatta ggtcacttaa caaaaaggaa attgggtagg ggtttttcac
     5281 agaccgcttt ctaagggtaa ttttaaaata tctgggaagt cccttccact gctgtgttcc
     5341 agaagtgttg gtaaacagcc cacaaatgtc aacagcagaa acatacaagc tgtcagcttt
     5401 gcacaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     5461 tta
//
