LOCUS       sequence-354640-        8639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for CRISPR-mediated knock-in line that expresses a QF2 
            transcriptional activator by knocking-in a T2A-QF2 fragment 
            replacing the stop codon of the endogenous Aedes aegypti Ir25a gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8639)
  AUTHORS   Herre M, Goldman OV, Lu TC, Caballero-Vidal G, Qi Y, Gilbert ZN, 
            Gong Z, Morita T, Rahiel S, Ghaninia M, Ignell R, Matthews BJ, Li H,
            Vosshall LB, Younger MA
  TITLE     Non-canonical odor coding in the mosquito.
  JOURNAL   Cell. 2022 Aug 18;185(17):3104-3123.e28. doi: 
            10.1016/j.cell.2022.07.024.
  PUBMED    35985288
REFERENCE   2  (bases 1 to 8639)
  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"; doi: 
            "10.1016/j.cell.2022.07"; journalName: "Cell"; date: "2022-08-18- 
            18"; volume: "185"; issue: "17"; pages: "3104-3123.e28"
COMMENT     Sequence Label: IR25a-T2A-QF2
FEATURES             Location/Qualifiers
     source          1..8639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2235..2288
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      3875..3884
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3881..4558
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(4061..4081)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     4473..4496
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    4679..4800
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4716..4735)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4770..4789
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6418..6434)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6418..6434)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6431..6453)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6442..6458
                     /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)."
     promoter        complement(join(6466..6472,6473..6490,6491..6496))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6511..6532
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6649..6666)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6820..7408)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6900..6919)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7579..8370,8371..8439))
                     /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     8202..8221
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8440..8544)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8612..8630
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggatcg
      421 agtttatact gtggtggtaa gttattaata ttcttaagga tattccaggc ctttcaaaaa
      481 aacattgatt gattaccttt acctacaatc tttacattgt ttcgtagcaa gaaccattca
      541 ttatacgaga tccaactgct cccaaaggtt tcaaaggtta ttgtatcgac ttgttggatg
      601 aaattgcaaa aattgttaag ttcgattatg aaatcaagga agttgaagat ggcaagtttg
      661 gaaacatgaa cgaaaaaggt gaatggaacg gtatcgtacg caaattgatc gacaagcaag
      721 ccgatatcgg tttaggatcg atgtccgtga tggccgagcg ggaaactgta attgatttca
      781 cggtgccata ttacgatttg gttggaatat caatcatgat gctgctgccc agtacaccga
      841 gttcgctgtt caaatttttg accgtgttgg agacgaatgt gtggctttgt attctcgcag
      901 cgtatttctt cactagtttt ctcatgtgga tttttgatcg gtacagtcct tacagttacc
      961 agaacaatcg tgaaaagtac aagaatgatg atgaaaaacg agagttcaac attaaggaat
     1021 gcctatggtt ttgtatgact tcgttaacgc cgcaaggagg tggtgaagct ccgaaaaatt
     1081 tatccggacg gctggtggcg gcaacgtggt ggttgtttgg gtaagtattc tactgaattt
     1141 tgtttagata ataaatattt aatggatgat ttatgtaaaa cataaagatt catcatcatt
     1201 gcatcgtaca ctgctaactt ggctgctttc ctgaccgtat cgcgattgga cactcccgtg
     1261 gaatcgttgg acgatctttc gaagcagtat aaaattttgt acgccccttt gaacggatct
     1321 tcggctatga cttatttcca gcgaatggcg gatatcgaag cccgtttcta tgagtaagtt
     1381 tgttctcatt catgcttact aggggtaatt ttctaagcaa aatcactctc cacagaattt
     1441 ggaaagagat gtcgctgaat gattctctca cacccgtgga gcgctcgaag ctggcggttt
     1501 gggactatcc cgtcagtgat aagtatacga aaatgtggca agcaatgcag gaagctggac
     1561 tcccaaatag tttggatgaa gcggtcgagc gtattaggaa ctccacatcg gcctctggtt
     1621 tcgcttttct gggcgatgct actgatatac gatacaaagt acttacaaac tgtgatctac
     1681 agatggtcgg cgaagaattt tctcgcaaac cgtatgccat tgcagtacag caaggatcgc
     1741 cgttgaaaga tcaattcaac aatgcgtaag tttagctgac ggatgtccta ttcaatataa
     1801 caacttaata taccattata ccttctagca tcttgatgct tctcaacaaa cgacaacttg
     1861 aaaaactcaa ggaacaatgg tggaaaaatg atgacattca aagcaaatgt gaaaaaccgg
     1921 acgaccaatc cgatggtatt tcgattcaga acattggagg tgtcttcata gtgatcttcg
     1981 tcggcattgg aatggcgtgc attacgctgg tctttgaatt ctggtactac aaataccgca
     2041 aaaacgtaaa gattattgat gtcgccgagg ctacggaaga taaactggct cagaagacgt
     2101 ccaatttgag gttgccaaat ttgaagaacg aatttggagc catgggacag atcaaagatg
     2161 acagtcagaa aacgcaaagt ttgcgaactc gtacgcagac gatagacgcc aacaacttta
     2221 aatctcgttt tggagagggc agaggaagtc ttctaacatg cggtgacgtg gaggagaatc
     2281 ccggccctat gccacccaag cgcaaaacgc ttaacgctgc ggctgaggct aacgctcatg
     2341 ccgacggaca cgccgacgga aacgccgacg gacacgtggc caatacggcc gcgtcctcga
     2401 ataatgcgag gttcgctgat ctcactaaca tcgatactcc gggtctggga cccacaacta
     2461 cgaccctgct cgtggaacca gcacgctcaa agcgtcaacg agtgtcccgc gcatgcgacc
     2521 agtgccgtgc agcccgagag aaatgcgacg gaatacagcc tgcgtgtttc ccgtgcgttt
     2581 cccagggaag gtcctgcact tatcaggctt cgccgaaaaa gaggggagtt caaaccggtt
     2641 atattcgtac gctggagctc gccctcgcct ggatgtttga aaatgtcgcg cgttccgaag
     2701 atgccttgca taacctcctc gtccgtgacg ccggacaagg atcagctctg ctcgttggta
     2761 aagattcgcc ggctgccgag cgactccatg cccgttgggc tactagccgt gtcaataaga
     2821 gcattacccg cctcctccgt cagttggagc tccctcctac cgccacggct acggcctcga
     2881 taatgccgca cgtgatggag cagcctctca gtaccagcat taaccccgtc aacgaccgct
     2941 tcaacggtat tcccaacccc actccgtata actccgatgc agctctcgat gctatcactc
     3001 agaccaacga ttatggaagc gtaaatacac atggtatcct ctctacttac ccgccaccgg
     3061 ctacgcacct taatgaagct tccgtcgctc tcgctcccgg tggcgccccc ccccgaccgc
     3121 ctcctccgta tgttgacagc acgaccaatc acccgccgta ccactcgaat ctggttccaa
     3181 tggcgaactt tggttactcg accgttgatt acgatgccat ggttgacgat ttggctagca
     3241 ttgaatacac ggacgctgtg gatgtcgacc cacagtttat gaccaatctg ggattcgttc
     3301 ctggatgtaa cttctccgac attaatacat acgaacagtg atgacgtcta tcgataccgt
     3361 cgactaaagc caaatagaaa attattcagt tcctggctta agtttttaaa agtgatatta
     3421 tttatttggt tgtaaccaac caaaagaatg taaataacta atacataatt atgttagttt
     3481 taagttagca acaaattgat tttagctata ttagctactt ggttaataaa tagaatatat
     3541 ttatttaaag ataattgcgt ttttattgtc agggagtgag tttgcttaaa aactcgttta
     3601 gatccaattc gagctcgccc ggggatctaa ttcaattaga gactaattca attagagcta
     3661 attcaattag gatccaagct tatcgatttc gaaccctcga ccgccggagt ataaatagag
     3721 gcgcttcgtc tacggagcga caattcaatt caaacaagca aagtgaacac gtcgctaagc
     3781 gaaagctaag caaataaaca agcgcagctg aacaagctaa acaatcgggg taccgctaga
     3841 gtcgacggta ccgcgggccc gggatccacc ggtcgccacc atggcctcct ccgagaacgt
     3901 catcaccgag ttcatgcgct tcaaggtgcg catggagggc accgtgaacg gccacgagtt
     3961 cgagatcgag ggcgagggcg agggccgccc ctacgagggc cacaacaccg tgaagctgaa
     4021 ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttccagta
     4081 cggctccaag gtgtacgtga agcaccccgc cgacatcccc gactacaaga agctgtcctt
     4141 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggcgaccgt
     4201 gacccaggac tcctccctgc aggacggctg cttcatctac aaggtgaagt tcatcggcgt
     4261 gaacttcccc tccgacggcc ccgtgatgca gaagaagacc atgggctggg aggcctccac
     4321 cgagcgcctg tacccccgcg acggcgtgct gaagggcgag acccacaagg ccctgaagct
     4381 gaaggacggc ggccactacc tggtggagtt caagtccatc tacatggcca agaagcccgt
     4441 gcagctgccc ggctactact acgtggacgc caagctggac atcacctccc acaacgagga
     4501 ctacaccatc gtggagcagt acgagcgcac cgagggccgc caccacctgt tcctgtagcg
     4561 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     4621 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     4681 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4741 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4801 gatactgaag ttcatttttt gagtagtaac gtgttgtttg aaattaccca atagagaaga
     4861 aacatatttt tgtctatcgc atattaacag gttccacttg aatttttaga tttcttcaat
     4921 tatttttttt tgtttcataa tcagaagaaa tatgaaaatt ggactggatc agttcttctt
     4981 tctcgactcg tatttttttc ttctcgaaaa aattggaaca ctatggaaaa acgcttaaga
     5041 tctctattct accctagttt aactagtaca ataaacatct tggctacatt aaatttattc
     5101 tgtggataaa tctgaacaca cattttctac atcgatttat actaggattc agacccacag
     5161 agtaatagaa atacaaccct gtggaattcg cattgtatcg taaagctagt gggctacaat
     5221 caaagtgtag tcgtaaaagg tctttttcga aaatttttga attgtataaa agttctcgca
     5281 aaagatctaa tatctagtat cccacacagg atgcgatgca tggaattcca acaacagcaa
     5341 aaaaaaacgc gtatcgaatt taagatacga ggggtagtaa cgtgttgtta tatcattctt
     5401 gatataatcg ttattttgag agtagtaacg tgttagtagg attggttatg taattttaat
     5461 acagtagcta gtaagatcat cccaaatcat ttaagggtgt attatggaaa ttgagtcgac
     5521 ctcatgagaa aaaagtcatc tcaatttaca tgattgtttc cttacaatgt ttgcatccaa
     5581 tccgaaattc cttaacaact tttcatacat tctcgagcac tttatgctta aaagccccga
     5641 aggctttccg gtctgtctcg ttcaacgtcc acgtttcatg tcggtaagaa gccaccagat
     5701 gtatccgctt ttcacgcatt ttgtttttat tttggggttg tggattttaa gctggttaca
     5761 aagttctccc accactacca agcggttgcc tggttctcta gatttgtgct cttgaaaatt
     5821 cgaggtttgg cttcgatgca tcatcacctt aatcttcata tgcgagataa actggttgaa
     5881 cgatccccca aaatcgtaga gatcctagac tatagagaac aatccaataa tgtatgttaa
     5941 agccgacgtc ggatagcgct tccaaaagaa ccacgagtgg gaaaaaagaa agtgaagaaa
     6001 atttgaaact ggcggtcaag aaatatccga aaccgaagac agtgtaacaa aagaacttag
     6061 gccgtcgtta ttaattggag gaagcggaat acgccaaaac cctgaagtcg atgcgaagca
     6121 ctgagaagct atccgcctaa tgacgacatc gtgcaaacta gcatcgatga aatgaagtta
     6181 aaaaaacaaa caaactagtg tagtctgttg tgtggaagag ccggaaggtg gcttggacga
     6241 ggtgctcaag aattggaggc caagatgctg agaacggggc tgaaggtgcg gatgtcggtt
     6301 gatcggacga cgagctcgga agtgctggag gaactcggaa gtcaagaggg gcttttctat
     6361 agtagggtac aaaaatctag agtcgacctg caggcatgca agcttggcgt aatcatggtc
     6421 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6481 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6541 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6601 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6661 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6721 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6781 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6841 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6901 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6961 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7021 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7081 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7141 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7201 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7261 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7321 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7381 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7441 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7501 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7561 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7621 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7681 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7741 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7801 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7861 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7921 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7981 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8041 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8101 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8161 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8221 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8281 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8341 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8401 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8461 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8521 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     8581 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
