LOCUS       sequence-342935-        4184 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac insect transgenesis destination vector for Golden Gate 
            Cloning assemblies.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4184)
  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 4184)
  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: pPiggyBacATCC LacZ GCTT loxAttP
FEATURES             Location/Qualifiers
     source          1..4184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..20
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     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)."
     protein_bind    663..684
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(699..704,705..722,723..729)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    737..753
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     742..764
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     761..777
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             773..1348
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
                     FPRQALNRGLPLGFRFSALRHLDPKKLD"
     primer_bind     796..816
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        798..816
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    829..936
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     853..869
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     853..869
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(903..919)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(904..920)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(944..963)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(945..963)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(973..989)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(982..1004)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1131..1511
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1218..1237)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1428..1449
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     protein_bind    complement(1580..1613)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   1801..1863
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(1993..2009)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      2174..2201
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2360..2377)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2531..3119)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2611..2630)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3266..4075)
                     /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     3989..4008
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4067..4086
                     /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 atccagagac ccgcaacgca
      661 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
      721 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
      781 gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga gctccaccgc
      841 ggtggcggcc gctctagaac tagtggatcc cccgggctgc aggaattcga tatcaagctt
      901 atcgataccg tcgacctcga gggggggccc ggtacccaat tcgccctata gtgagtcgta
      961 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     1021 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     1081 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     1141 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     1201 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1261 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1321 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     1381 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     1441 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     1501 gcctatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgggtct
     1561 ccgcttgtaa ttcgtcgaca taacttcgta tagcatacat tatacgaagt tatgagctca
     1621 attcgataaa agttttgtta ctttatagaa gaaattttga gtttttgttt tttttaataa
     1681 ataaataaac ataaataaat tgtttgttga atttattatt agtatgtaag tgtaaatata
     1741 ataaaactta atatctattc aaattaataa ataaacctcg atatacagac cgataaaaca
     1801 catgcgtcaa ttttacgcat gattatcttt aacgtacgtc acaatatgat tatctttcta
     1861 gggttaatct agctgcgtgt tctgcagcgt gtcgagcatc ttcatctgct ccatcacgct
     1921 gtaaaacaca tttgcaccgc gagtctgccc gtcctccacg ggttcaaaaa cgtgaatgaa
     1981 cgaggcgcgc tcactggccg tcgttttaca ggggatgtct tcatatatat gaagactccc
     2041 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     2101 aacgttgtga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     2161 catcaaacta agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt
     2221 cctggctagc ggtacgcgta ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2281 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2341 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2401 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2461 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2521 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2581 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2641 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2701 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2761 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2821 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2881 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2941 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3001 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3061 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3121 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgacgcg taactcacgt
     3181 taagggattt tggtcatgag cttgcgccgt cccgtcaagt cagcgtaatg ctctgccagt
     3241 gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca
     3301 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     3361 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     3421 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     3481 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt
     3541 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     3601 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa
     3661 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     3721 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt tttccgggga
     3781 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     3841 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     3901 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaagcgat
     3961 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     4021 catccatgtt ggaatttaat cgcggcctcg acgtttcccg ttgaatatgg ctcataacac
     4081 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat
     4141 cttgtgcaat gtaacatcag agattttgag acacgggcca gagc
//
