LOCUS       sequence-243711-        5329 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes firefly luciferase under the pIEx promoter for constitutive 
            expression in insect (e.g. Drosophila, mosquito) cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5329)
  AUTHORS   Fredericks AC, Russell TA, Wallace LE, Davidson AD, Fernandez-Sesma 
            A, Maringer K
  TITLE     Aedes aegypti (Aag2)-derived clonal mosquito cell lines reveal the 
            effects of pre-existing persistent infection with the 
            insect-specific bunyavirus Phasi Charoen-like virus on arbovirus 
            replication.
  JOURNAL   PLoS Negl Trop Dis. 2019 Nov 6;13(11):e0007346. doi: 
            10.1371/journal.pntd.0007346. eCollection 2019 Nov.
  PUBMED    31693659
REFERENCE   2  (bases 1 to 5329)
  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: "2019-11-6"; volume: "13(11):e0007346. doi"; 
            pages: " 10.1371/journal.pntd.0007346. eCollection 2019 Nov"
COMMENT     Sequence Label: pKM19
FEATURES             Location/Qualifiers
     source          1..5329
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        486..1077
                     /label=IE1 promoter
                     /note="promoter of the ie1 gene from the baculovirus 
                     Autographa californica"
     CDS             1165..2817
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(1231..1249)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      2845..3152
                     /label=IE1 terminator
                     /note="terminator of the ie1 gene from the baculovirus 
                     Autographa californica"
     promoter        3182..3286
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3287..3355,3356..4147)
                     /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(3505..3524)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4318..4906
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4807..4826
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5060..5077
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5194..5215
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5230..5235,5236..5253,5254..5260)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5268..5284
                     /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)."
     primer_bind     5273..5295
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5292..5308
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5292..5308
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        join(5329,1..482)
                     /gene="Autographa californica hr5"
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
ORIGIN
        1 gcgtaaaaca caatcaagta tgagtcataa gctgatgtca tgttttgcac acggctcata
       61 accgaactgg ctttacgagt agaattctac ttgtaacgca cgatcagtgg atgatgtcat
      121 ttgtttttca aatcgagatg atgtcatgtt ttgcacacgg ctcataaact cgctttacga
      181 gtagaattct acgtgtaacg cacgatcgat tgatgagtca tttgttttgc aatatgatat
      241 catacaatat gactcatttg tttttcaaaa ccgaacttga tttacgggta gaattctact
      301 tgtaaagcac aatcaaaaag atgatgtcat ttgtttttca aaactgaact cgctttacga
      361 gtagaattct acgtgtaaaa cacaatcaag aaatgatgtc atttgttata aaaataaaag
      421 ctgatgtcat gttttgcaca tggctcataa ctaaactcgc tttacgggta gaattctacg
      481 cgcgtcgatg tctttgtgat gcgcgcgaca tttttgtagg ttattgataa aatgaacgga
      541 tacgttgccc gacattatca ttaaatcctt ggcgtagaat ttgtcgggtc cattgtccgt
      601 gtgcgctagc atgcccgtaa cggacctcgt acttttggct tcaaaggttt tgcgcacaga
      661 caaaatgtgc cacacttgca gctctgcatg tgtgcgcgtt accacaaatc ccaacggcgc
      721 agtgtacttg ttgtatgcaa ataaatctcg ataaaggcgc ggcgcgcgaa tgcagctgat
      781 cacgtacgtc cctcgtgttc cgttcaagga cggtgttatc gacctcagat taatgtttat
      841 cggccgactg ttttcgtatc cgctcaccaa acgcgttttt gcattaacat tgtatgtcgg
      901 cggatgttct atatctaatt tgaataaata aacgataacc gcgttggttt tagagggcat
      961 aataaaagaa atattgttat cgtgttcgcc attagggcag tataaattga cgttcatgtt
     1021 ggatattgtt tcagttgcaa gttgacactg gcggcgacaa gatcgtgaac aaccaagtga
     1081 ccatggagat ctaattgcgg ccgcaattat gcatcaacca tgggagctcg aattccagct
     1141 tggcattccg gtactgttgg taaaatggaa gacgccaaaa acataaagaa aggcccggcg
     1201 ccattctatc ctctagagga tggaaccgct ggagagcaac tgcataaggc tatgaagaga
     1261 tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt gaacatcacg
     1321 tacgcggaat acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg
     1381 aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg
     1441 ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt
     1501 gaattgctca acagtatgaa catttcgcag cctaccgtag tgtttgtttc caaaaagggg
     1561 ttgcaaaaaa ttttgaacgt gcaaaaaaaa ttaccaataa tccagaaaat tattatcatg
     1621 gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta
     1681 cctcccggtt ttaatgaata cgattttgta ccagagtcct ttgatcgtga caaaacaatt
     1741 gcactgataa tgaattcctc tggatctact gggttaccta agggtgtggc ccttccgcat
     1801 agaactgcct gcgtcagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt
     1861 ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca
     1921 ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagagctg
     1981 tttttacgat cccttcagga ttacaaaatt caaagtgcgt tgctagtacc aaccctattt
     2041 tcattcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt
     2101 gcttctgggg gcgcacctct ttcgaaagaa gtcggggaag cggttgcaaa acgcttccat
     2161 cttccaggga tacgacaagg atatgggctc actgagacta catcagctat tctgattaca
     2221 cccgaggggg atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag
     2281 gttgtggatc tggataccgg gaaaacgctg ggcgttaatc agagaggcga attatgtgtc
     2341 agaggaccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt
     2401 gacaaggatg gatggctaca ttctggagac atagcttact gggacgaaga cgaacacttc
     2461 ttcatagttg accgcttgaa gtctttaatt aaatacaaag gatatcaggt ggcccccgct
     2521 gaattggaat cgatattgtt acaacacccc aacatcttcg acgcgggcgt ggcaggtctt
     2581 cccgacgatg acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg
     2641 atgacggaaa aagagatcgt ggattacgtc gccagtcaag taacaaccgc gaaaaagttg
     2701 cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca
     2761 agaaaaatca gagagatcct cataaaggcc aagaagggcg gaaagtccaa attgtaactc
     2821 gagcactaag tgattaacct caggttatac atatattttg aatttaatta attatacata
     2881 tattttatat tatttttgtc ttttattatc gaggggccgt tgttggtgtg gggttttgca
     2941 tagaaataac aatgggagtt ggcgacgttg ctgcgccaac accacctccc ttccctcctt
     3001 tcatcatgta tctgtagata aaataaaata ttaaacctaa aaacaagacc gcgcctatca
     3061 acaaaatgat aggcattaac ttgccgctga cgctgtcact aacgttggac gatttgccga
     3121 ctaaaccttc atcgcccagt aaccaatcta gacgtcaggt ggcacttttc ggggaaatgt
     3181 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3241 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3301 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3361 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3421 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3481 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3541 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3601 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3661 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3721 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3781 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3841 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3901 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3961 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4021 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4081 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4141 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     4201 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4261 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4321 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4381 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4441 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4501 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4561 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4621 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4681 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4741 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4801 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4861 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4921 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4981 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     5041 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     5101 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     5161 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     5221 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     5281 acaatttcac acaggaaaca gctatgacca tgattacgaa ttcccgggc
//
