LOCUS       sequence-361316-        7924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac plasmid for insect transformation, expressing PhiC31 
            integrase under control of the Aedes aegypti PUb promoter, GFP 
            fluorescent marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7924)
  AUTHORS   Lutrat C, Burckbuchler M, Olmo RP, Beugnon R, Fontaine A, Akbari OS,
            Argiles-Herrero R, Baldet T, Bouyer J, Marois E
  TITLE     Combining two genetic sexing strains allows sorting of 
            non-transgenic males for Aedes genetic control.
  JOURNAL   Commun Biol. 2023 Jun 16;6(1):646. doi: 10.1038/s42003-023-05030-7.
  PUBMED    37328568
REFERENCE   2  (bases 1 to 7924)
  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: "Commun 
            Biol."; date: "2023-06-16"; volume: "6(1):646. doi"; pages: " 
            10.1038/s42003-023-05030-7"
COMMENT     Sequence Label: pB PUb-integrase
FEATURES             Location/Qualifiers
     source          1..7924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..20
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4..20
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   61..95
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     CDS             1781..1801
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1817..3631
                     /codon_start=1
                     /product="integrase from phage phi-C31"
                     /label=phage phi-C31 integrase
                     /translation="DTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDGG
                     RFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIPI
                     VSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQRELG
                     GYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWREI
                     KTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRILR
                     DPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQAW
                     LDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPGQ
                     HEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERAN
                     LVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAEA
                     PKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPIE
                     KRASITWAKPPTDDDEDDAQDGTEDVAA"
     polyA_signal    3739..3821
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      4369..4378
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4375..4377,4378..4380,4381..5094)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4375..4377,4378..4380,4381..5091)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4420..4441)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4681..4700)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5028..5049
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5216..5297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   5541..5603
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5733..5750)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5733..5749)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5914..5941
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(6100..6117)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6271..6859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6351..6370)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7006..7815)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     7729..7748
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7807..7826
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tgccaggaaa cagctatgac catgtaatac gacgatatga tcctgatgca gctagattaa
       61 ccctagaaag atagtctgcg taaaattgac gcatgcattc ttgaaatatt gctctctctt
      121 tctaaatagc gcgaatccgt cgctgtgcat ttaggacatc tcagtcgccg cttggagctc
      181 ccgtgaggcg tgcttgtcaa tgcggtaagt gtcactgatt ttgaactata acgaccgcgt
      241 gagtcaaaat gacgcatgat tatcttttac gtgactttta agatttaact catacgataa
      301 ttatattgtt atttcatgtt ctacttacgt gataacttat tatatatata ttttcttgtt
      361 atagattaga tcgcgctcgc ggatccatct ttacatgtag cttgtgcatt gaatccaatt
      421 ataatttgcc ttggcaccag ctgagccaga caagaaagaa agcttcccag aagtatatcg
      481 atttagaagg gttgacgtca ctttgctgac tgcactaata cagcaaatga tacaattaga
      541 atgattcaag tgaaattccc aaattactgc tttgtctctg gatttggtta tcagattaca
      601 ttcgaagcta agaatagcta ccgaaattgt cgatcaaatc aggaaatcct ttctctatcg
      661 aaaaaggcat tcgcacatct ttctctgtat gccatataca cgaatggtag gtacattgac
      721 gtctttgcca gaagttgaac tgcatcgttc aaggtacaga atgaacgact aacagacaca
      781 agcacgtttt gctgtccatt cagacacagg gatggtaccc atagtcgatc gatttagagc
      841 catccaaccg aacagaggta tatgtatgaa tggattgcag aaattttcta gaagtacaac
      901 caccactacg gcagtgtcta taaaacgccc ctgcaaaggc aaaaccagct caatcgaata
      961 cgtttcctag tggagtgaac attacgcggt ccaagtaagc agtgccagtg caagtgaagt
     1021 gaagtctcta gtgaaaaaga gtgatccaat tagccagagg agaaaatttc agagtgaaca
     1081 aagctttgtt caaaggacaa ttactattaa atttgtgaaa gtgcatttcg gtgaagggaa
     1141 tcttctagtg aaggtaggta aattaaatga tgaaattata gctatgagcg aaaactagtt
     1201 tggtgaatga ttcctttgtc tttgaatgag caaactattt tccaagatgg cgactattga
     1261 gctttgagtg attagtgaaa atttgcaacg cagtttcatc atcattgata aaacccaatt
     1321 gtgattcacg gcgataatca tatttcgttg aatcatcgct gctaattgaa ttaaatttct
     1381 agagcaagcg cgaattcgcc atatttctaa aattaaatat tgtggtgata attacccatt
     1441 aaggtaatat taacacatat cgagaaaaac cttgaggaaa tcgtgaaaac ttgaagatac
     1501 gcaatttcca aactacgtag ttcaaagtcg aaaacaagtt aatttttcac taaaaagtag
     1561 ggcgttgttg tgacgtcatc accttcaagt gtatattttt cacttggcct gcgactgcaa
     1621 acgcagacaa agcaaaacaa gtttaaaacc tgtcgtgtcg tgctcgaagc caaaggcaat
     1681 gaatcaatat caaatgagag tttgcatttc acaaccaatt actgaagcgt ttcctcgttt
     1741 ctttttctgc tcaacagaga tttcaacaat gagcggccct ccaaaaaaga agagaaaggt
     1801 agaagacccg ggcggcgaca cgtacgcggg tgcttacgac cgtcagtcgc gcgagcgcga
     1861 aaattcgagc gcagcaagcc cagcgacaca gcgtagcgcc aacgaagaca aggcggccga
     1921 ccttcagcgc gaagtcgagc gcgacggggg ccggttcagg ttcgtcgggc atttcagcga
     1981 agcgccgggc acgtcggcgt tcgggacggc ggagcgcccg gagttcgaac gcatcctgaa
     2041 cgaatgccgc gccgggcggc tcaacatgat cattgtctat gacgtgtcgc gcttctcgcg
     2101 cctgaaggtc atggacgcga ttccgattgt ctcggaattg ctcgccctgg gcgtgacgat
     2161 tgtttccact caggaaggcg tcttccggca gggaaacgtc atggacctga ttcacctgat
     2221 tatgcggctc gacgcgtcgc acaaagaatc ttcgctgaag tcggcgaaga ttctcgacac
     2281 gaagaacctt cagcgcgaat tgggcgggta cgtcggcggg aaggcgcctt acggcttcga
     2341 gcttgtttcg gagacgaagg agatcacgcg caacggccga atggtcaatg tcgtcatcaa
     2401 caagcttgcg cactcgacca ctccccttac cggacccttc gagttcgagc ccgacgtaat
     2461 ccggtggtgg tggcgtgaga tcaagacgca caaacacctt cccttcaagc cgggcagtca
     2521 agccgccatt cacccgggca gcatcacggg gctttgtaag cgcatggacg ctgacgccgt
     2581 gccgacccgg ggcgagacga ttgggaagaa gaccgcttca agcgcctggg acccggcaac
     2641 cgttatgcga atccttcggg acccgcgtat tgcgggcttc gccgctgagg tgatctacaa
     2701 gaagaagccg gacggcacgc cgaccacgaa gattgagggt taccgcattc agcgcgaccc
     2761 gatcacgctc cggccggtcg agcttgattg cggaccgatc atcgagcccg ctgagtggta
     2821 tgagcttcag gcgtggttgg acggcagggg gcgcggcaag gggctttccc gggggcaagc
     2881 cattctgtcc gccatggaca agctgtactg cgagtgtggc gccgtcatga cttcgaagcg
     2941 cggggaagaa tcgatcaagg actcttaccg ctgccgtcgc cggaaggtgg tcgacccgtc
     3001 cgcacctggg cagcacgaag gcacgtgcaa cgtcagcatg gcggcactcg acaagttcgt
     3061 tgcggaacgc atcttcaaca agatcaggca cgccgaaggc gacgaagaga cgttggcgct
     3121 tctgtgggaa gccgcccgac gcttcggcaa gctcactgag gcgcctgaga agagcggcga
     3181 acgggcgaac cttgttgcgg agcgcgccga cgccctgaac gcccttgaag agctgtacga
     3241 agaccgcgcg gcaggcgcgt acgacggacc cgttggcagg aagcacttcc ggaagcaaca
     3301 ggcagcgctg acgctccggc agcaaggggc ggaagagcgg cttgccgaac ttgaagccgc
     3361 cgaagccccg aagcttcccc ttgaccaatg gttccccgaa gacgccgacg ctgacccgac
     3421 cggccctaag tcgtggtggg ggcgcgcgtc agtagacgac aagcgcgtgt tcgtcgggct
     3481 cttcgtagac aagatcgttg tcacgaagtc gactacgggc agggggcagg gaacgcccat
     3541 cgagaagcgc gcttcgatca cgtgggcgaa gccgccgacc gacgacgacg aagacgacgc
     3601 ccaggacggc acggaagacg tagcggcgta gtaagcataa tcagccatac cacatttgta
     3661 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     3721 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3781 agcatcacaa atttcacaaa taaagcattt ttcttcactg cattctagtt gtggtttgtc
     3841 caaactcatc aatgtatcta agtatctaat tcaattagag actaattcaa ttagagtcta
     3901 attcaattag agttatctaa ttcaattaga gactaattca attagagtct aattcaatta
     3961 gagttatcta attcaattag agactaattc aattagagtc taattcaatt agagttatct
     4021 aattcaatta gagactaatt caattagagt ctaattcaat tagagttatc taattcaatt
     4081 agagactaat tcaattagag tctaattcaa ttagagttat ctaattcaat tagagactaa
     4141 ttcaattaga gctaattcaa ttaggatcca agcttatcga tttcgaaccc tcgaccgccg
     4201 gagtataaat agaggcgctt cgtctacgga gcgacaattc aattcaaaca agcaaagtga
     4261 acacgtcgct aagcgaaagc taagcaaata aacaagcgca gctgaacaag ctaaacaatc
     4321 ggggtaccgc tagagtcgac ggtaccgcgg gcccgggatc caccggtcgc caccatggtg
     4381 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     4441 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     4501 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     4561 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     4621 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     4681 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     4741 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     4801 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     4861 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     4921 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     4981 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     5041 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
     5101 cgcgactcta gatcaaatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     5161 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     5221 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5281 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcgcttgt
     5341 aattcgtcga catgagctca attcgataaa agttttgtta ctttatagaa gaaattttga
     5401 gtttttgttt tttttaataa ataaataaac ataaataaat tgtttgttga atttattatt
     5461 agtatgtaag tgtaaatata ataaaactta atatctattc aaattaataa ataaacctcg
     5521 atatacagac cgataaaaca catgcgtcaa ttttacgcat gattatcttt aacgtacgtc
     5581 acaatatgat tatctttcta gggttaatct agctgcgtgt tctgcagcgt gtcgagcatc
     5641 ttcatctgct ccatcacgct gtaaaacaca tttgcaccgc gagtctgccc gtcctccacg
     5701 ggttcaaaaa cgtgaatgaa cgaggcgcgc tcactggccg tcgttttaca ggggatgtct
     5761 tcatatatat gaagactccc atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     5821 atccgccggg agcggatttg aacgttgtga agcaacggcc cggagggtgg cgggcaggac
     5881 gcccgccata aactgccagg catcaaacta agcagaaggc catcctgacg gatggccttt
     5941 ttgcgtttct acaaactctt cctggctagc ggtacgcgta ttaattgcgt tgcgctcact
     6001 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6061 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6121 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6181 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6241 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6301 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6361 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6421 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6481 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6541 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6601 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6661 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6721 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6781 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6841 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6901 gaacgacgcg taactcacgt taagggattt tggtcatgag cttgcgccgt cccgtcaagt
     6961 cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc
     7021 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     7081 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     7141 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     7201 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     7261 tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     7321 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     7381 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     7441 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     7501 gaatgctgtt tttccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     7561 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     7621 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     7681 gggcttccca tacaagcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     7741 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg acgtttcccg
     7801 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     7861 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacgggcca
     7921 gagc
//
