LOCUS       sequence-361370-        4443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  attB-site containing docking plasmid for genomic insertion into attP
            site, encoding GFP under control of the Aedes aegypti PUb promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4443)
  AUTHORS   Lutrat C, Burckbuchler M, Olmo RP, Beugnon R, Fontaine A, Akbari OS,
            Argiles-Herrero R, Baldet T, Bouyer J, Marois E
  TITLE     Combining two genetic sexing strains allows sorting of 
            non-transgenic males for Aedes genetic control.
  JOURNAL   Commun Biol. 2023 Jun 16;6(1):646. doi: 10.1038/s42003-023-05030-7.
  PUBMED    37328568
REFERENCE   2  (bases 1 to 4443)
  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: "2023-06-16"; volume: "6(1):646. doi"; pages: " 
            10.1038/s42003-023-05030-7"
COMMENT     Sequence Label: pENTR-attB-PUb-GFP
FEATURES             Location/Qualifiers
     source          1..4443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             join(1661..1663,1664..1666,1667..2380)
                     /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(1661..1663,1664..1666,1667..2377)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1706..1727)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1967..1986)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2314..2335
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2502..2583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2539..2558)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2593..2612
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    2630..2663
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     primer_bind     complement(2684..2703)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2685..2703)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2708..2724)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2708..2724)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2826..2845)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2837..3646
                     /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     complement(2904..2923)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3793..4381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4282..4301
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggcatccat ctttacatgt agcttgtgca
      301 ttgaatccaa ttataatttg ccttggcacc agctgagcca gacaagaaag aaagcttccc
      361 agaagtatat cgatttagaa gggttgacgt cactttgctg actgcactaa tacagcaaat
      421 gatacaatta gaatgattca agtgaaattc ccaaattact gctttgtctc tggatttggt
      481 tatcagatta cattcgaagc taagaatagc taccgaaatt gtcgatcaaa tcaggaaatc
      541 ctttctctat cgaaaaaggc attcgcacat ctttctctgt atgccatata cacgaatggt
      601 aggtacattg acgtctttgc cagaagttga actgcatcgt tcaaggtaca gaatgaacga
      661 ctaacagaca caagcacgtt ttgctgtcca ttcagacaca gggatggtac ccatagtcga
      721 tcgatttaga gccatccaac cgaacagagg tatatgtatg aatggattgc agaaattttc
      781 tagaagtaca accaccacta cggcagtgtc tataaaacgc ccctgcaaag gcaaaaccag
      841 ctcaatcgaa tacgtttcct agtggagtga acattacgcg gtccaagtaa gcagtgccag
      901 tgcaagtgaa gtgaagtctc tagtgaaaaa gagtgatcca attagccaga ggagaaaatt
      961 tcagagtgaa caaagctttg ttcaaaggac aattactatt aaatttgtga aagtgcattt
     1021 cggtgaaggg aatcttctag tgaaggtagg taaattaaat gatgaaatta tagctatgag
     1081 cgaaaactag tttggtgaat gattcctttg tctttgaatg agcaaactat tttccaagat
     1141 ggcgactatt gagctttgag tgattagtga aaatttgcaa cgcagtttca tcatcattga
     1201 taaaacccaa ttgtgattca cggcgataat catatttcgt tgaatcatcg ctgctaattg
     1261 aattaaattt ctagagcaag cgcgaattcg ccatatttct aaaattaaat attgtggtga
     1321 taattaccca ttaaggtaat attaacacat atcgagaaaa accttgagga aatcgtgaaa
     1381 acttgaagat acgcaatttc caaactacgt agttcaaagt cgaaaacaag ttaatttttc
     1441 actaaaaagt agggcgttgt tgtgacgtca tcaccttcaa gtgtatattt ttcacttggc
     1501 ctgcgactgc aaacgcagac aaagcaaaac aagtttaaaa cctgtcgtgt cgtgctcgaa
     1561 gccaaaggca atgaatcaat atcaaatgag agtttgcatt tcacaaccaa ttactgaagc
     1621 gtttcctcgt ttctttttct gctcaacaga gatttcaaca atggtgagca agggcgagga
     1681 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1741 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     1801 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     1861 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1921 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1981 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2041 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     2101 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     2161 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2221 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     2281 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     2341 cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg actctagatc
     2401 aaatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     2461 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2521 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2581 tgcattctag ttgtggtttg tccaaactca tcaatgtatc gctttgcggg tgccagggcg
     2641 tgcccttggg ctccccgggc gcgtactcca cctcacccat atcccctata gtgagtcgta
     2701 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2761 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2821 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2881 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2941 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     3001 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     3061 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     3121 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     3181 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     3241 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     3301 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     3361 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     3421 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     3481 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     3541 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     3601 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     3661 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3721 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3781 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3841 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3901 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3961 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4021 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4081 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4141 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4201 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4261 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4321 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4381 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4441 gtt
//
