LOCUS       sequence-342937-        9035 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 9035)
  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 9035)
  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: ppBAalbNixE1E2E3E4 PUb-DsRed
FEATURES             Location/Qualifiers
     source          1..9035
                     /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    3978..4060
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             join(5494..5496,5497..5499,5500..6174)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(5677..5697)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     5895..5914
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6089..6112
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    6295..6376
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(6431..6464)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   6652..6714
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6844..6861)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6844..6860)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      7025..7052
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(7211..7228)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7382..7970)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7462..7481)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8117..8926)
                     /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     8840..8859
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8918..8937
                     /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 caaggtatgg cagaattgaa aaaaatagac tagtgtacga ctcaaccgga
     3361 cactctaaac aatacggttt tgtttattat gaaaagcact tgtctgctca agcggccaaa
     3421 caggaaatgg accgcagtga tcatacagga cgtaaaattg cactgcggac cgttcaagaa
     3481 attttggcag cggaaacatt tcttgacctt ttccacaata cttatcagct ttggaaaatc
     3541 cccagaggta gatcaatcct actccgccaa aaacttccct ggcgatggaa gctcttccga
     3601 accgatcttc ccgagtcggt gtcgagctag attctgtttt ttccggttgg agactgatct
     3661 ccggttcact ggcgccccag cgaccgtgct ttccatgacg ttatcgatct agcctactct
     3721 cccggcggtg gacctagcct actctgatgg aaaaaatccc cagcgaaaga agttctcccg
     3781 aaaccgagaa ttcaatctcg accatccagg tgccgaggtc gacccagcga tgccggttgg
     3841 aggttgactt ccagttcact atcactccga taagcataat cagccatacc acatttgtag
     3901 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     3961 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4021 gcatcacaaa tttcacaaat aaagcatttt tcttcactgc attctagttg tggtttgtcc
     4081 aaactcatca atgtatctaa gagttatctt tacatgtagc ttgtgcattg aatccaatta
     4141 taatttgcct tggcaccagc tgagccagac aagaaagaaa gcttcccaga agtatatcga
     4201 tttagaaggg ttgacgtcac tttgctgact gcactaatac agcaaatgat acaattagaa
     4261 tgattcaagt gaaattccca aattactgct ttgtctctgg atttggttat cagattacat
     4321 tcgaagctaa gaatagctac cgaaattgtc gatcaaatca ggaaatcctt tctctatcga
     4381 aaaaggcatt cgcacatctt tctctgtatg ccatatacac gaatggtagg tacattgacg
     4441 tctttgccag aagttgaact gcatcgttca aggtacagaa tgaacgacta acagacacaa
     4501 gcacgttttg ctgtccattc agacacaggg atggtaccca tagtcgatcg atttagagcc
     4561 atccaaccga acagaggtat atgtatgaat ggattgcaga aattttctag aagtacaacc
     4621 accactacgg cagtgtctat aaaacgcccc tgcaaaggca aaaccagctc aatcgaatac
     4681 gtttcctagt ggagtgaaca ttacgcggtc caagtaagca gtgccagtgc aagtgaagtg
     4741 aagtctctag tgaaaaagag tgatccaatt agccagagga gaaaatttca gagtgaacaa
     4801 agctttgttc aaaggacaat tactattaaa tttgtgaaag tgcatttcgg tgaagggaat
     4861 cttctagtga aggtaggtaa attaaatgat gaaattatag ctatgagcga aaactagttt
     4921 ggtgaatgat tcctttgtct ttgaatgagc aaactatttt ccaagatggc gactattgag
     4981 ctttgagtga ttagtgaaaa tttgcaacgc agtttcatca tcattgataa aacccaattg
     5041 tgattcacgg cgataatcat atttcgttga atcatcgctg ctaattgaat taaatttcta
     5101 gagcaagcgc gaattcgcca tatttctaaa attaaatatt gtggtgataa ttacccatta
     5161 aggtaatatt aacacatatc gagaaaaacc ttgaggaaat cgtgaaaact tgaagatacg
     5221 caatttccaa actacgtagt tcaaagtcga aaacaagtta atttttcact aaaaagtagg
     5281 gcgttgttgt gacgtcatca ccttcaagtg tatatttttc acttggcctg cgactgcaaa
     5341 cgcagacaaa gcaaaacaag tttaaaacct gtcgtgtcgt gctcgaagcc aaaggcaatg
     5401 aatcaatatc aaatgagagt ttgcatttca caaccaatta ctgaagcgtt tcctcgtttc
     5461 tttttctgct caacagagat ttcaacgttc accatggtgc gctcctccaa gaacgtcatc
     5521 aaggagttca tgcgcttcaa ggtgcgcatg gagggcaccg tgaacggcca cgagttcgag
     5581 atcgagggcg agggcgaggg ccgcccctac gagggccaca acaccgtgaa gctgaaggtg
     5641 accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc
     5701 tccaaggtgt acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc
     5761 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     5821 caggactcct ccctgcagga cggctgcttc atctacaagg tgaagttcat cggcgtgaac
     5881 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag
     5941 cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc acaaggccct gaagctgaag
     6001 gacggcggcc actacctggt ggagttcaag tccatctaca tggccaagaa gcccgtgcag
     6061 ctgcccggct actactacgt ggactccaag ctggacatca cctcccacaa cgaggactac
     6121 accatcgtgg agcagtacga gcgcaccgag ggccgccacc acctgttcct gtagcggccg
     6181 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     6241 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     6301 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6361 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcgcttgta
     6421 attcgtcgac ataacttcgt atagcataca ttatacgaag ttatgagctc aattcgataa
     6481 aagttttgtt actttataga agaaattttg agtttttgtt ttttttaata aataaataaa
     6541 cataaataaa ttgtttgttg aatttattat tagtatgtaa gtgtaaatat aataaaactt
     6601 aatatctatt caaattaata aataaacctc gatatacaga ccgataaaac acatgcgtca
     6661 attttacgca tgattatctt taacgtacgt cacaatatga ttatctttct agggttaatc
     6721 tagctgcgtg ttctgcagcg tgtcgagcat cttcatctgc tccatcacgc tgtaaaacac
     6781 atttgcaccg cgagtctgcc cgtcctccac gggttcaaaa acgtgaatga acgaggcgcg
     6841 ctcactggcc gtcgttttac aggggatgtc ttcatatata tgaagactcc catctgttgt
     6901 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgtg
     6961 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaact
     7021 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tcctggctag
     7081 cggtacgcgt attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     7141 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     7201 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     7261 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     7321 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7381 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7441 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7501 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7561 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7621 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7681 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7741 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7801 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     7861 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7921 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7981 tgatcttttc tacggggtct gacgctcagt ggaacgacgc gtaactcacg ttaagggatt
     8041 ttggtcatga gcttgcgccg tcccgtcaag tcagcgtaat gctctgccag tgttacaacc
     8101 aattaaccaa ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     8161 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     8221 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     8281 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     8341 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     8401 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     8461 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     8521 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     8581 cacctgaatc aggatattct tctaatacct ggaatgctgt ttttccgggg atcgcagtgg
     8641 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     8701 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     8761 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaagcga tagattgtcg
     8821 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     8881 tggaatttaa tcgcggcctc gacgtttccc gttgaatatg gctcataaca ccccttgtat
     8941 tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa
     9001 tgtaacatca gagattttga gacacgggcc agagc
//
