LOCUS       sequence-342953-        9462 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 9462)
  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 9462)
  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 PUb-GFP
FEATURES             Location/Qualifiers
     source          1..9462
                     /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"
     CDS             join(5881..5883,5884..5886,5887..6600)
                     /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(5881..5883,5884..5886,5887..6597)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5926..5947)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6187..6206)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6534..6555
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6722..6803
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(6858..6891)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   7079..7141
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7271..7288)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7271..7287)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      7452..7479
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(7638..7655)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7809..8397)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7889..7908)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8544..9353)
                     /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     9267..9286
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9345..9364
                     /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 tatctaagag ttatctttac
     4501 atgtagcttg tgcattgaat ccaattataa tttgccttgg caccagctga gccagacaag
     4561 aaagaaagct tcccagaagt atatcgattt agaagggttg acgtcacttt gctgactgca
     4621 ctaatacagc aaatgataca attagaatga ttcaagtgaa attcccaaat tactgctttg
     4681 tctctggatt tggttatcag attacattcg aagctaagaa tagctaccga aattgtcgat
     4741 caaatcagga aatcctttct ctatcgaaaa aggcattcgc acatctttct ctgtatgcca
     4801 tatacacgaa tggtaggtac attgacgtct ttgccagaag ttgaactgca tcgttcaagg
     4861 tacagaatga acgactaaca gacacaagca cgttttgctg tccattcaga cacagggatg
     4921 gtacccatag tcgatcgatt tagagccatc caaccgaaca gaggtatatg tatgaatgga
     4981 ttgcagaaat tttctagaag tacaaccacc actacggcag tgtctataaa acgcccctgc
     5041 aaaggcaaaa ccagctcaat cgaatacgtt tcctagtgga gtgaacatta cgcggtccaa
     5101 gtaagcagtg ccagtgcaag tgaagtgaag tctctagtga aaaagagtga tccaattagc
     5161 cagaggagaa aatttcagag tgaacaaagc tttgttcaaa ggacaattac tattaaattt
     5221 gtgaaagtgc atttcggtga agggaatctt ctagtgaagg taggtaaatt aaatgatgaa
     5281 attatagcta tgagcgaaaa ctagtttggt gaatgattcc tttgtctttg aatgagcaaa
     5341 ctattttcca agatggcgac tattgagctt tgagtgatta gtgaaaattt gcaacgcagt
     5401 ttcatcatca ttgataaaac ccaattgtga ttcacggcga taatcatatt tcgttgaatc
     5461 atcgctgcta attgaattaa atttctagag caagcgcgaa ttcgccatat ttctaaaatt
     5521 aaatattgtg gtgataatta cccattaagg taatattaac acatatcgag aaaaaccttg
     5581 aggaaatcgt gaaaacttga agatacgcaa tttccaaact acgtagttca aagtcgaaaa
     5641 caagttaatt tttcactaaa aagtagggcg ttgttgtgac gtcatcacct tcaagtgtat
     5701 atttttcact tggcctgcga ctgcaaacgc agacaaagca aaacaagttt aaaacctgtc
     5761 gtgtcgtgct cgaagccaaa ggcaatgaat caatatcaaa tgagagtttg catttcacaa
     5821 ccaattactg aagcgtttcc tcgtttcttt ttctgctcaa cagagatttc aacgttcacc
     5881 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     5941 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     6001 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     6061 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     6121 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     6181 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     6241 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     6301 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     6361 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     6421 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     6481 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     6541 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     6601 agcggccgcg actctagatc aaatcagcca taccacattt gtagaggttt tacttgcttt
     6661 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     6721 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6781 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6841 gcttgtaatt cgtcgacata acttcgtata gcatacatta tacgaagtta tgagctcaat
     6901 tcgataaaag ttttgttact ttatagaaga aattttgagt ttttgttttt tttaataaat
     6961 aaataaacat aaataaattg tttgttgaat ttattattag tatgtaagtg taaatataat
     7021 aaaacttaat atctattcaa attaataaat aaacctcgat atacagaccg ataaaacaca
     7081 tgcgtcaatt ttacgcatga ttatctttaa cgtacgtcac aatatgatta tctttctagg
     7141 gttaatctag ctgcgtgttc tgcagcgtgt cgagcatctt catctgctcc atcacgctgt
     7201 aaaacacatt tgcaccgcga gtctgcccgt cctccacggg ttcaaaaacg tgaatgaacg
     7261 aggcgcgctc actggccgtc gttttacagg ggatgtcttc atatatatga agactcccat
     7321 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     7381 cgttgtgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca
     7441 tcaaactaag cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactcttcc
     7501 tggctagcgg tacgcgtatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     7561 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     7621 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     7681 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7741 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7801 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7861 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7921 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7981 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     8041 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     8101 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     8161 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     8221 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     8281 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     8341 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     8401 gatcctttga tcttttctac ggggtctgac gctcagtgga acgacgcgta actcacgtta
     8461 agggattttg gtcatgagct tgcgccgtcc cgtcaagtca gcgtaatgct ctgccagtgt
     8521 tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat
     8581 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
     8641 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
     8701 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
     8761 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct
     8821 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     8881 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa
     8941 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
     9001 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt tccggggatc
     9061 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
     9121 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     9181 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caagcgatag
     9241 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     9301 tccatgttgg aatttaatcg cggcctcgac gtttcccgtt gaatatggct cataacaccc
     9361 cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct
     9421 tgtgcaatgt aacatcagag attttgagac acgggccaga gc
//
