LOCUS       sequence-472661-        9155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac plasmid for the insertion of a sperm-expressed green 
            fluorescent protein in Aedes aegypti.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9155)
  TITLE     Mosquito sex under lock and key
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9155)
  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: 
            "Unpublished"
COMMENT     Sequence Label: PB{3xP3-dsRed2; beta-2Tubulin-mNeonGreen}
FEATURES             Location/Qualifiers
     source          1..9155
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   1172..1206
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    2622..2743
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(2864..3574)
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      3310..3319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3571..3580
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4582..4703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4824..5501)
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMSFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(4886..4909)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     5301..5321
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     regulatory      5498..5507
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   6764..6826
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7530..7547)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7530..7546)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7539..7561)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7755..7774)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7874..7896
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7934..7952)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8020..8124
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8125..8193,8194..8985)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8343..8362)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga catgattacg aattcgagct cggtacccgg
     1021 ggatcctcta gagtcgacgc tcgcgcgact tggtttgcca ttctttagcg cgcgtcgcgt
     1081 cacacagctt ggccacaatg tggtttttgt caaacgaaga ttctatgacg tgtttaaagt
     1141 ttaggtcgag taaagcgcaa atctttttta accctagaaa gatagtctgc gtaaaattga
     1201 cgcatgcatt cttgaaatat tgctctctct ttctaaatag cgcgaatccg tcgctgtgca
     1261 tttaggacat ctcagtcgcc gcttggagct cccgtgaggc gtgcttgtca atgcggtaag
     1321 tgtcactgat tttgaactat aacgaccgcg tgagtcaaaa tgacgcatga ttatctttta
     1381 cgtgactttt aagatttaac tcatacgata attatattgt tatttcatgt tctacttacg
     1441 tgataactta ttatatatat attttcttgt tatagatatc gtgactaata tataataaaa
     1501 tgggtagttc tttagacgat gagcatatcc tctctgctct tctgcaaagc gatgacgagc
     1561 ttgttggtga ggattctgac agtgaaatat cagatcacgt aagtgaagat gacgtccaga
     1621 gcgatacaga agaagcgttt atagatgagg tacatgaagt gcagccaacg tcaagcggta
     1681 gtgaaatatt agacgaacaa aatgttattg aacaaccagg ttcttcattg gcttctaaca
     1741 gaatcttgac cttgccacag aggactatta gaggtaagaa taaacattgt tggtcaactt
     1801 caaagtccac gaggcgtagc cgagtctctg cactgaacat tgtcagatct caaagaggtc
     1861 cgacgcgtat gtgccgcaat atatatgacc cacttttatg cttcaaacta ttttttactg
     1921 atgagataat ttcggaaatt gtaaaatgga caaatgctga gatatcattg aaacgtcggg
     1981 aatctatgac aggtgctaca tttcgtgaca cgaatgaaga tgaaatctat gctttctttg
     2041 gtattctggt aatgacagca gtgagaaaag ataaccacat gtccacagat gacctctttg
     2101 atcgatcttt gtcaatggtg tacgtctctg taatgagtcg tgatcgtttt gattttttga
     2161 tacgatgtct tagaatggat gacaaaagta tacggcccac acttcgagaa aacgatgtat
     2221 ttactcctgt tagaaaaata tgggatctct ttatccatca gtgcatacaa aattacactc
     2281 caggggctca tttgaccata gatgaacagt tacttggttt tagaggacgg tgtccgttta
     2341 ggatgtatat cccaaacaag ccaagtaagt atggaataaa aatcctcatg atgtgtgaca
     2401 gtggtacgaa gtatatgata aatggaatgc cttatttggg aagaggaaca cagaccaacg
     2461 gagtaccact cggtgaatac tacgtgaagg agttatcaaa gcctgtgcac ggtagttgtc
     2521 gtaatattac gtgtgacaat tggttcacct caatcccttt ggcaaaaaac ttactacaag
     2581 aaccgtataa gttcgagatc ggccggccta ggcgcgcccc ttaagataca ttgatgagtt
     2641 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2701 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2761 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     2821 ctacaaatgt ggtatggctg attatgatct agagtcgcgg ccgttacttg tacagctcgt
     2881 ccatgcccat cacatcggta aaggcctttt gccactcctt gaagttgagc tcggtcttgg
     2941 agtgcttgag ctccgtctta cggaacacgt acatcggctg gttcttcaga tagttagccg
     3001 ccattggctt ggcaaaggtg taggtggtcc gcgcagtgct ccggtagcgc ttgccatttc
     3061 cagtggtgta actccactta aaggtactga tgatggtttt gtcgttgggg taagtcttct
     3121 tcgacctgca ccagtccgca gcggtcagcg agttggtcat cacaggaccg tcagcaggga
     3181 aaccagtccc cttcacctgg gcctctcctt tgatgtggct tccctcgtag gtgtagcggt
     3241 agttaacagt aagggaggca ccatcttcaa actgcattgt gcgatggact tggtagccgg
     3301 agccatctac catggcggcc tggaaaggcg acatcccgtc agggtagggc aggtactgat
     3361 ggaagccata cccgatatga gggaccagaa tccaggggga gaactggagg tcacccttgg
     3421 tggacttcag gtttaactcc tcataaccat catttggatt gccggtgccc tgacccacca
     3481 tgtcaaagtc cacaccgttg atggagccaa agatgtgtaa ctcatgtgtc gctgggagag
     3541 aggccatgtt atcctcctcg cccttgctca ccatggtggc gaccggtgga tcccctggag
     3601 cacttctagc ggttccaaaa ttgggcttca aaatatgctt cttccgatgc ttgaagatgg
     3661 aagcagaatg gttccaaatc acgtccgcgt tcctcggtgg tgaaaatcga agtgaaaatt
     3721 tttccaactg tcgtcctccg ggtcgtcaag aagaaggatg gccttccatg acacactgtg
     3781 attgatttac attgtggcgg cagtatgggc ccccaaaacg ggagcgacgc gttcaacatg
     3841 tgctactgga tgcattcgct tcgaagtctg ttgtcacgtt gggggctgtc catatgccac
     3901 gtggttgtta tggggtggag aagggtactg ctcaagcatc acggtgcata caaaatttga
     3961 aaatttttgt gtggacgaaa gatcataagg aggagcgtga aaattcataa aaaatgacca
     4021 cgtggtttat ggacgaaaca aaagcgatat atgagccaca agaacatagc gtgattcacc
     4081 cccacattag gggaaacatg caagtccttt gcataaacta tcaaaatttg gcggaaaata
     4141 atagtgttga aatgaataat aacagttaca actaggtatt agttcttcct cttttcggca
     4201 ttacgtctcc tttgtggcag agcatgtttc tgagtttaaa aaccatcatt tattttttgt
     4261 tgattgctga tgattgaaga ttgaatgata gatttttgaa ccttaaaaca aaaaagttaa
     4321 cgcgatttaa actattatga ggggtttaaa aattctaacg ttcagagtgt ctactaattt
     4381 acagaaatga aattccctga tatttctagg ttttccaggc ttggaaaaaa atccaggttt
     4441 aagaaagttt acaacataca caaattattg atgaattttt cattttagat aagtagtaat
     4501 cattttcaaa gctaggagtt ttgcttcgaa atacttaaac aaaaaatcta aggggcgcgc
     4561 cgtacgcgta tcgataagct ttaagataca ttgatgagtt tggacaaacc acaactagaa
     4621 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     4681 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     4741 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg
     4801 attatgatct agagtcgcgg ccgctacagg aacaggtggt ggcggccctc ggtgcgctcg
     4861 tactgctcca cgatggtgta gtcctcgttg tgggaggtga tgtccagctt ggcgtccacg
     4921 tagtagtagc cgggcagctg cacgggcttc ttggccatgt agatggactt gaactccacc
     4981 aggtagtggc cgccgtcctt cagcttcagg gccttgtggg tctcgccctt cagcacgccg
     5041 tcgcgggggt acaggcgctc ggtggaggcc tcccagccca tggtcttctt ctgcatcacg
     5101 gggccgtcgg aggggaagtt cacgccgatg aacttcacct tgtagatgaa gcagccgtcc
     5161 tgcagggagg agtcctgggt cacggtcgcc acgccgccgt cctcgaagct catcacgcgc
     5221 tcccacttga agccctcggg gaaggacagc ttcttgtagt cggggatgtc ggcggggtgc
     5281 ttcacgtaca ccttggagcc gtactggaac tggggggaca ggatgtccca ggcgaagggc
     5341 agggggccgc ccttggtcac cttcagcttc acggtgttgt ggccctcgta ggggcggccc
     5401 tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg tgccctccat gcgcaccttg
     5461 aagcgcatga actcggtgat gacgttctcg gaggaggcca tggtggcgac cggtggatcc
     5521 cgggcccgcg gtaccgtcga ctctagcggt accccgattg tttagcttgt tcagctgcgc
     5581 ttgtttattt gcttagcttt cgcttagcga cgtgttcact ttgcttgttt gaattgaatt
     5641 gtcgctccgt agacgaagcg cctctattta tactccggcg gtcgagggtt cgaaatcgat
     5701 aagcttggat cctaattgaa ttagctctaa ttgaattagt ctctaattga attagatccc
     5761 cgggcgagct cgaattaacc attgtgggaa ccgtgcgatc aaacaaacgc gagataccgg
     5821 aagtactgaa aaacagtcgc tccaggccag tgggaacatc gatgttttgt tttgacggac
     5881 cccttactct cgtctcatat aaaccgaagc cagctaagat ggtatactta ttatcatctt
     5941 gtgatgagga tgcttctatc aacgaaagta ccggtaaacc gcaaatggtt atgtattata
     6001 atcaaactaa aggcggagtg gacacgctag accaaatgtg ttctgtgatg acctgcagta
     6061 ggaagacgaa taggtggcct atggcattat tgtacggaat gataaacatt gcctgcataa
     6121 attcttttat tatatacagc cataatgtca gtagcaaggg agaaaaggtt caaagtcgca
     6181 aaaaatttat gagaaacctt tacatgagcc tgacgtcatc gtttatgcgt aagcgtttag
     6241 aagctcctac tttgaagaga tatttgcgcg ataatatctc taatattttg ccaaatgaag
     6301 tgcctggtac atcagatgac agtactgaag agccagtaat gaaaaaacgt acttactgta
     6361 cttactgccc ctctaaaata aggcgaaagg caaatgcatc gtgcaaaaaa tgcaaaaaag
     6421 ttatttgtcg agagcataat attgatatgt gccaaagttg tttctgactg actaataagt
     6481 ataatttgtt tctattatgt ataagttaag ctaattactt attttataat acaacatgac
     6541 tgtttttaaa gtacaaaata agtttatttt tgtaaaagag agaatgttta aaagttttgt
     6601 tactttatag aagaaatttt gagtttttgt ttttttttaa taaataaata aacataaata
     6661 aattgtttgt tgaatttatt attagtatgt aagtgtaaat ataataaaac ttaatatcta
     6721 ttcaaattaa taaataaacc tcgatataca gaccgataaa acacatgcgt caattttacg
     6781 catgattatc tttaacgtac gtcacaatat gattatcttt ctagggttaa ataatagttt
     6841 ctaatttttt tattattcag cctgctgtcg tgaataccgt atatctcaac gctgtctgtg
     6901 agattgtcgt attctagcct ttttagtttt tcgctcatcg acttgatatt gtccgacaca
     6961 ttttcgtcga tttgcgtttt gatcaaagac ttgagcagag acacgttaat caactgttca
     7021 aattgatcca tattaacgat atcaacccga tgcgtatatg gtgcgtaaaa tatatttttt
     7081 aaccctctta tactttgcac tctgcgttaa tacgcgttcg tgtacagacg taatcatgtt
     7141 ttcttttttg gataaaactc ctactgagtt tgacctcata ttagaccctc acaagttgca
     7201 aaacgtggca ttttttacca atgaagaatt taaagttatt ttaaaaaatt tcatcacaga
     7261 tttaaagaag aaccaaaaat taaattattt caacagttta atcgaccagt taatcaacgt
     7321 gtacacagac gcgtcggcaa aaaacacgca gcccgacgtg ttggctaaaa ttattaaatc
     7381 aacttgtgtt atagtcacgg atttgccgtc caacgtgttc ctcaaaaagt tgaagaccaa
     7441 caagtttacg gacactatta attatttgat tttgccccac ttcattttgt gggatcacaa
     7501 ttttgttata tttttaaaca aagcttggca ctggccgtcg ttttacaacg tcgtgactgg
     7561 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     7621 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     7681 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     7741 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     7801 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     7861 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     7921 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     7981 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     8041 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     8101 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     8161 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     8221 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     8281 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     8341 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     8401 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     8461 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     8521 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     8581 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     8641 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     8701 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     8761 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     8821 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     8881 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     8941 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     9001 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     9061 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     9121 gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
