LOCUS       sequence-342936-        8766 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transgenesis vector for Aedes albopictus ectopic expression of the 
            Nix gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8766)
  AUTHORS   Lutrat C, Olmo RP, Baldet T, Bouyer J, Marois E
  TITLE     Transgenic expression of Nix converts genetic females into males and
            allows automated sex sorting in Aedes albopictus.
  JOURNAL   Commun Biol. 2022 Mar 7;5(1):210. doi: 10.1038/s42003-022-03165-7.
  PUBMED    35256751
REFERENCE   2  (bases 1 to 8766)
  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: "2022-03-7"; volume: "5(1):210. doi"; pages: " 
            10.1038/s42003-022-03165-7"
COMMENT     Sequence Label: ppBAalbNixE1E2 OpIE2-GFP
FEATURES             Location/Qualifiers
     source          1..8766
                     /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)"
     protein_bind    427..526
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     protein_bind    complement(602..635)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             4070..4090
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     polyA_signal    4365..4447
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4497..5044
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 promoter
                     /note="strong constitutive baculovirus promoter for insect 
                     cell expression"
     promoter        5080..5127
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        5119..5137
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(5147..5149,5150..5152,5153..5866)
                     /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(5147..5149,5150..5152,5153..5863)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5192..5213)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5453..5472)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5800..5821
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5988..6069
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(6162..6195)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   6383..6445
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6575..6592)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6575..6591)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      6756..6783
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(6942..6959)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7113..7701)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7193..7212)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7848..8657)
                     /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     8571..8590
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8649..8668
                     /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 gcgactgacg gtcgtaagca cccgcgtacg tgtccacccc
      421 ggtcacaacc ccttgtgtca tgtcggcgac cctacgcccc caactgagag aactcaaagg
      481 ttaccccagt tggggcacta ctcccgaaaa ccgcttctga cctgggaaaa cgtgaagccc
      541 cggggcatcc gctgagggtt gccgccgggg cttcggtgtg tccgtcagta ctgcaggtac
      601 cataacttcg tatagcatac attatacgaa gttataccgg atcctcgcat tttatgagta
      661 aaggcccatt actcatatat gggtaaagtg ctttttgtaa aaaatgagta aatcgattta
      721 ctcataaatt ataatttcac cgtgtttact tttcgtcttt tgttatattt ttgaactgtt
      781 aactattttg tatttatcat ttaattcata caaaattttg cgtttatttt gtagtttatt
      841 gcaatctacg aaaattatta atgaattttt acatgtttca aatactctta atcattttcc
      901 tcctggaaaa cagaccttcc gagacgtagc tctttgaagt tttcataatg atccatgtat
      961 gaaattttcc cctccggctg ttgttcatcg cgcagttgtt cctgttcttc cgttcctcgt
     1021 tcttccggcg cttgaagcct tggtgcagtc ggaactgtgg aataagactg atatattttt
     1081 atttcacggt cgttaatgaa aatgatgctg tcactatgga acttacctgc aaattccatg
     1141 cttttgtagc ctttcttgta gctccacatg atgatttgtt gctcctgtat tatcttcgag
     1201 taccaaacct tgtgaaacgg aaataatcca tcgtaaatga aaattgaacg gtatcaattt
     1261 atccaggaag tacttaccga ttaaaatatc aatgtatttg ggaacctcga ctctaatccg
     1321 cgtgggaatc cgatggacgc ggtatcggtc attgaacccg atgctgaagg gcctcgactg
     1381 gctatagcgg tgccgtgaaa atcaaaatat ctaaaatttc ccgatagtgt tatgatcagg
     1441 ggatccgcat gcggcaaggt aggcctactg ctcatctttt gcgagtgtgt tagtgtgact
     1501 ggcaacgcga ccagaagtct agtcgaatcc tgcttggaaa tcggtttgtt ttgtcgatgg
     1561 gggtggcccg cgaaatcgca ttgtgttgtt tgggaaggcc cgctcgatgc atgttaggta
     1621 ggcgtagttt gttcgagttt gacgtgcctg caccctccct ggttatgatg gcgcactaat
     1681 gcattccgag aaaagttgct aacttgaaaa ttttcatcca aatatgggta agttcattta
     1741 cccatgtttg ggtatagcga ttttacctat aatatgggca aatttgaccg tttttcaaag
     1801 gtatatttta cccatattat gggtaatgca tatgaaaccc aaatatgggt aaatcaacta
     1861 ctgatctatg ggtaaactga tcttagcgtg tatgtttcac acgtgacatt cactggtttt
     1921 acaattggct tctttagcag tgttgggata attccatatt atgaatctgc atgccaaact
     1981 gggccgaaat ccaaattttc atcaattttg gtgcacggga acctatttaa atatcaattt
     2041 gaagtttgta tgggagcgat ttgtcgaatc acccctcgtt gcattttgta ctgggcggag
     2101 ctgtcaaaca gttgcctagc tgtcaaaagg tgatttcgaa taatctcttt gaaattgatt
     2161 ttaggtatca aaataaagtt ctaaaaatct gaaaaaaatc atagtggctc agaaaaaggt
     2221 gctctttcgt ataaaatcaa aaaatgaaca cttttttcaa aatttaaaaa cccaattttc
     2281 gcaaatctat agggctctgc acgaagttct ctccctctct ttcgctctca ttgagatttt
     2341 gtaaacaaca aggccaggaa atgtcaaaat cccatacaaa atcaaaacag tgcagtgccc
     2401 tatatgtaaa acacatcact ccgacgtgta aatttttttg agtgttgatt taatcaaagt
     2461 gaataaaaat attagtttta tgacatactt gttttctgag tgtagcaaaa atatgaaaac
     2521 acatttttgt actttgaatg ttaagcgtgt atgctttttg gtttggttat gtcaatatgt
     2581 caattgttaa acccatgtta atagttttaa ttttttttaa tcaaattctt tttttaagta
     2641 atgtacagta aaagtgaact taatctcatt aacaatcaat ttgaatacat taaaaaatat
     2701 tgcatataca ttggaaacat tcccgccgaa gtatcgaaaa cagatttaat tgcaaaattt
     2761 tccgtatttg gtgaaatatc taacttatac atgaagtcat tcattcagtt ttgtgatgtg
     2821 aaaccggcag ttgttcgtta cagactgatg aaaagtgtaa aggaatcttc aagtttacac
     2881 aatagtcgat atattcaatc ggttttaata gttctgccac tagattcttc ctacaataat
     2941 tactttcttc cttacaacac ttgtgttgtg gtatacactt ataacaaatt tggcatggta
     3001 gatttttatc aaaaattcag taaattagga gatatacatg cgatgaagaa agctacaaat
     3061 gtcatggttt acattagctt tgtatcagaa agagctgcaa ggaccattct ggatactaag
     3121 cctacagata tacatataaa tgtacaaaca attaatcatg ttacacgaaa tattaacgta
     3181 tgcttaatag attttgaaaa ggaatgtaca tcaaatacgg cgataaaatt aacactttta
     3241 tataatcgct caattggaat attcggacta ccatctaatt tcacagaagc aaaactgcac
     3301 gatgaatttt caaggtttgt gtggcaatgt gaatccaaat gagacagata aatataaaag
     3361 ctactcagat gaaaagacct tgaaggtcga aattagacat tatattttta tatgttttca
     3421 ggtatggcag aattgaaaaa aatagactag tgtacgactc aaccggacac tctaaacaat
     3481 acggttttgt ttattatgaa aagcacttgt ctgctcaagc ggccaaacag gaaatggacc
     3541 gcagtgatca tacaggacgt aaaattgcag ttcggtttgt tccagaaaaa gagtagggca
     3601 gatattaaat ctagtgacag aacaaagcaa ctagcaccgt actaaaaatg gcgtagggtt
     3661 ctcgaatatt tcgagagcta cttaccattt gtgtcataca caattaaaat gactgacaag
     3721 tctatctact tactcgcaat ctattttatc gcaggtttat caataagaat tttagtgcct
     3781 gcacaacgtt gggtttgatc tgatgtaata atcataatca aaattttcca accaaattaa
     3841 agctagctat atgtaaagta gccttgcgtg aaacaagtaa cttttgcgtg atttcataag
     3901 cccaaggctc aatttagcaa tcaatgatta gtactactat taatataacc atgttaggat
     3961 cggttttaaa taactaaatt tgttgcttct ggtgaaaaac catgtttgaa ctttatgtta
     4021 tattttgctg tcgggcttag ttaccgtgct acaatggatg gtgaaccttc atcatcacca
     4081 tcatcatcat aacagactga aagcatgaaa attctgctct taggattcac accatcgaaa
     4141 tcatcaataa ttagatatct tttcatctta tcagatagaa agatgttgga atatgctgaa
     4201 tgacatttcg aagtgtcaaa aaaccacacc gtagtatcac acggagcgca taaagcgtaa
     4261 gcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     4321 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     4381 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcatttttct
     4441 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatctaagta tcgatccatg
     4501 atgataaaca atgtatggtg ctaatgttgc ttcaacaaca attctgttga actgtgtttt
     4561 catgtttgcc aacaagcacc tttatactcg gtggcctccc caccaccaac ttttttgcac
     4621 tgcaaaaaaa cacgcttttg cacgcgggcc catacatagt acaaactcta cgtttcgtag
     4681 actattttac ataaatagtc tacaccgttg tatacgctcc aaatacacta ccacacattg
     4741 aacctttttg cagtgcaaaa aagtacgtgt cggcagtcac gtaggccggc cttatcgggt
     4801 cgcgtcctgt cacgtacgaa tcacattatc ggaccggacg agtgttgtct tatcgtgaca
     4861 ggacgccagc ttcctgtgtt gctaaccgca gccggacgca actccttatc ggaacaggac
     4921 gcgcctccat atcagccgcg cgttatctca tgcgcgtgac cggacacgag gcgcccgtcc
     4981 cgcttatcgc gcctataaat acagcccgca acgatctggt aaacacagtt gaacagcatc
     5041 tgttcgaatt aattcgggat ctgccgggct gcagcacgtg ttgacaatta atcatcggca
     5101 tagtatatcg gcatagtata atacgactca ctataggagg gccacaatgg tgagcaaggg
     5161 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     5221 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     5281 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     5341 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     5401 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     5461 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     5521 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     5581 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     5641 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     5701 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     5761 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     5821 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc
     5881 tagatcaaat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     5941 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     6001 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6061 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcgctt gtaattcgtc
     6121 gacgcggata tcgtttaaac cgttgcttgt aattcgtcga cataacttcg tatagcatac
     6181 attatacgaa gttatgagct caattcgata aaagttttgt tactttatag aagaaatttt
     6241 gagtttttgt tttttttaat aaataaataa acataaataa attgtttgtt gaatttatta
     6301 ttagtatgta agtgtaaata taataaaact taatatctat tcaaattaat aaataaacct
     6361 cgatatacag accgataaaa cacatgcgtc aattttacgc atgattatct ttaacgtacg
     6421 tcacaatatg attatctttc tagggttaat ctagctgcgt gttctgcagc gtgtcgagca
     6481 tcttcatctg ctccatcacg ctgtaaaaca catttgcacc gcgagtctgc ccgtcctcca
     6541 cgggttcaaa aacgtgaatg aacgaggcgc gctcactggc cgtcgtttta caggggatgt
     6601 cttcatatat atgaagactc ccatctgttg tttgtcggtg aacgctctcc tgagtaggac
     6661 aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg cccggagggt ggcgggcagg
     6721 acgcccgcca taaactgcca ggcatcaaac taagcagaag gccatcctga cggatggcct
     6781 ttttgcgttt ctacaaactc ttcctggcta gcggtacgcg tattaattgc gttgcgctca
     6841 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6901 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6961 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7021 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7081 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7141 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7201 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7261 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7321 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7381 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7441 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7501 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     7561 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7621 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7681 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7741 tggaacgacg cgtaactcac gttaagggat tttggtcatg agcttgcgcc gtcccgtcaa
     7801 gtcagcgtaa tgctctgcca gtgttacaac caattaacca attctgatta gaaaaactca
     7861 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     7921 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     7981 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     8041 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     8101 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     8161 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     8221 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     8281 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     8341 tggaatgctg tttttccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     8401 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     8461 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     8521 tcgggcttcc catacaagcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     8581 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct cgacgtttcc
     8641 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     8701 gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg agacacgggc
     8761 cagagc
//
