LOCUS       sequence-361358-        8967 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 8967)
  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 8967)
  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: ppBAalbNixE1E3E4 PUb-YFP
FEATURES             Location/Qualifiers
     source          1..8967
                     /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)."
     polyA_signal    3830..3912
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             join(5346..5348,5349..5351,5352..6062)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5391..5412)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5652..5671)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5999..6020
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6072..6092
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6227..6308
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(6363..6396)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   6584..6646
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6776..6793)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6776..6792)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      6957..6984
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(7143..7160)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7314..7902)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7394..7413)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8049..8858)
                     /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     8772..8791
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8850..8869
                     /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 caagctgcgg accgttcaag aaattttggc agcggaaaca tttcttgacc
     3361 ttttccacaa tacttatcag ctttggaaaa tccccagagg tagatcaatc ctactccgcc
     3421 aaaaacttcc ctggcgatgg aagctcttcc gaaccgatct tcccgagtcg gtgtcgagct
     3481 agattctgtt ttttccggtt ggagactgat ctccggttca ctggcgcccc agcgaccgtg
     3541 ctttccatga cgttatcgat ctagcctact ctcccggcgg tggacctagc ctactctgat
     3601 ggaaaaaatc cccagcgaaa gaagttctcc cgaaaccgag aattcaatct cgaccatcca
     3661 ggtgccgagg tcgacccagc gatgccggtt ggaggttgac ttccagttca ctatcactcc
     3721 gataagcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3781 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3841 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3901 tttcttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct aagagttatc
     3961 tttacatgta gcttgtgcat tgaatccaat tataatttgc cttggcacca gctgagccag
     4021 acaagaaaga aagcttccca gaagtatatc gatttagaag ggttgacgtc actttgctga
     4081 ctgcactaat acagcaaatg atacaattag aatgattcaa gtgaaattcc caaattactg
     4141 ctttgtctct ggatttggtt atcagattac attcgaagct aagaatagct accgaaattg
     4201 tcgatcaaat caggaaatcc tttctctatc gaaaaaggca ttcgcacatc tttctctgta
     4261 tgccatatac acgaatggta ggtacattga cgtctttgcc agaagttgaa ctgcatcgtt
     4321 caaggtacag aatgaacgac taacagacac aagcacgttt tgctgtccat tcagacacag
     4381 ggatggtacc catagtcgat cgatttagag ccatccaacc gaacagaggt atatgtatga
     4441 atggattgca gaaattttct agaagtacaa ccaccactac ggcagtgtct ataaaacgcc
     4501 cctgcaaagg caaaaccagc tcaatcgaat acgtttccta gtggagtgaa cattacgcgg
     4561 tccaagtaag cagtgccagt gcaagtgaag tgaagtctct agtgaaaaag agtgatccaa
     4621 ttagccagag gagaaaattt cagagtgaac aaagctttgt tcaaaggaca attactatta
     4681 aatttgtgaa agtgcatttc ggtgaaggga atcttctagt gaaggtaggt aaattaaatg
     4741 atgaaattat agctatgagc gaaaactagt ttggtgaatg attcctttgt ctttgaatga
     4801 gcaaactatt ttccaagatg gcgactattg agctttgagt gattagtgaa aatttgcaac
     4861 gcagtttcat catcattgat aaaacccaat tgtgattcac ggcgataatc atatttcgtt
     4921 gaatcatcgc tgctaattga attaaatttc tagagcaagc gcgaattcgc catatttcta
     4981 aaattaaata ttgtggtgat aattacccat taaggtaata ttaacacata tcgagaaaaa
     5041 ccttgaggaa atcgtgaaaa cttgaagata cgcaatttcc aaactacgta gttcaaagtc
     5101 gaaaacaagt taatttttca ctaaaaagta gggcgttgtt gtgacgtcat caccttcaag
     5161 tgtatatttt tcacttggcc tgcgactgca aacgcagaca aagcaaaaca agtttaaaac
     5221 ctgtcgtgtc gtgctcgaag ccaaaggcaa tgaatcaata tcaaatgaga gtttgcattt
     5281 cacaaccaat tactgaagcg tttcctcgtt tctttttctg ctcaacagag atttcaacgt
     5341 tcaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     5401 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     5461 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     5521 ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
     5581 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     5641 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     5701 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     5761 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     5821 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     5881 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     5941 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     6001 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     6061 agagatctcg acccaagaaa aagcggaagg tggaggaccc gtaaagcggc cgcgactcta
     6121 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     6181 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     6241 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6301 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcgcttg taattcgtcg
     6361 acataacttc gtatagcata cattatacga agttatgagc tcaattcgat aaaagttttg
     6421 ttactttata gaagaaattt tgagtttttg ttttttttaa taaataaata aacataaata
     6481 aattgtttgt tgaatttatt attagtatgt aagtgtaaat ataataaaac ttaatatcta
     6541 ttcaaattaa taaataaacc tcgatataca gaccgataaa acacatgcgt caattttacg
     6601 catgattatc tttaacgtac gtcacaatat gattatcttt ctagggttaa tctagctgcg
     6661 tgttctgcag cgtgtcgagc atcttcatct gctccatcac gctgtaaaac acatttgcac
     6721 cgcgagtctg cccgtcctcc acgggttcaa aaacgtgaat gaacgaggcg cgctcactgg
     6781 ccgtcgtttt acaggggatg tcttcatata tatgaagact cccatctgtt gtttgtcggt
     6841 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg tgaagcaacg
     6901 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ctaagcagaa
     6961 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttcctggct agcggtacgc
     7021 gtattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     7081 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     7141 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     7201 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     7261 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7321 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7381 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7441 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7501 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7561 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7621 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7681 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7741 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7801 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7861 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7921 tctacggggt ctgacgctca gtggaacgac gcgtaactca cgttaaggga ttttggtcat
     7981 gagcttgcgc cgtcccgtca agtcagcgta atgctctgcc agtgttacaa ccaattaacc
     8041 aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     8101 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     8161 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     8221 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     8281 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
     8341 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     8401 cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca
     8461 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     8521 tcaggatatt cttctaatac ctggaatgct gtttttccgg ggatcgcagt ggtgagtaac
     8581 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     8641 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     8701 ttcagaaaca actctggcgc atcgggcttc ccatacaagc gatagattgt cgcacctgat
     8761 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     8821 aatcgcggcc tcgacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     8881 atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat
     8941 cagagatttt gagacacggg ccagagc
//
