LOCUS       sequence-372758-        8502 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 Ir8a gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8502)
  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 8502)
  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: IR8a-T2A-QF2
FEATURES             Location/Qualifiers
     source          1..8502
                     /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             1991..2044
                     /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      3631..3640
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3637..4314
                     /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(3817..3837)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     4229..4252
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    4435..4556
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4472..4491)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4526..4545
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6281..6297)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6281..6297)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6294..6316)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6305..6321
                     /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(6329..6335,6336..6353,6354..6359))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6374..6395
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6512..6529)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6683..7271)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6763..6782)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7442..8233,8234..8302))
                     /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     8065..8084
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8303..8407)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8475..8493
                     /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 acccggtcag
      421 aatcttctca agattctcat cagaatcttc tcacgaatct cattagaatc ctctcaggat
      481 tatcattaga atcctctcaa gattctcatt agaatcctcc caggattctc atcagaatcc
      541 tctcaggaat ctcatcacac tgctctaaag atactcgtca gaatcctctt aggatcctca
      601 tcagaatcct ctcaggactc tcatcagaat cctctcagca ttctcatcag aatcccctca
      661 gcattctcat cgaaatcctt caggatgctc atcagaatcc tctaaagatt ctgatgagct
      721 tctttagagg attctcatca gaatcctctg aggattctca ttagaatcct ctcagcattc
      781 tctaaggatt tttttcagaa tcctctcagg attttcatta gaatcctgtc agaattttca
      841 ttagaatcct ctcaggattc ttatcagaac cctctcagga ttcagtgaag actttaacct
      901 ttaattctgt ttcagagact tcactctcaa ggaaagcttc tggtttgcct taacttcatt
      961 cactccacaa ggcggtggag aagccccaaa ggcactttcc ggacgtaccc tggtggcggc
     1021 atactggttg ttcgtcgtac taatgttggc tacatttacc gcgaacctgg ctgcgtttct
     1081 taccgttgaa cgtatgcaga ctccggtaca atccttggag cagctgtccc gtcaaagtcg
     1141 catcaaatac acagttgtga aggactccga cacgcatgat tacttcaaga acatgaagaa
     1201 cgccgaagac gtactgtacc gaatgtggcg taatctgacg ctttcgagtg gaaacgacca
     1261 gacgcagtat agcgtttggg attacccgat caaggaacag tataccaata tcttggctgc
     1321 catagagtca gcgaatccag tccagtcggc tgaagttgga ttccaacgag tcaacgaggg
     1381 tctagaagcg gattttgcat tcgttcacga ttcagcggag atcaagtacg aaatatcacg
     1441 caactgtaat ctcacggaag ttggggaagt tttcggtgaa cagccgtatg cgatcgcagt
     1501 tcaacaggga agtcatctgc aagacgagtt gagttattac attctggaac tgcaaaagga
     1561 acgctacttc gaatcgctga cggcaaagtt ttggaacaat tcggtacgcg gaatttgtcc
     1621 caatacggac gacagcgaag gaataacact ggaaagcttg ggtggagtgt tcatagcgac
     1681 tcttgtggga ctagccctgg caatggtaac tttggctggg gaagtgttgt attaccgccg
     1741 taaggatcag aagggcaagc tggtgcaggt tgagccgatg gatgtaggaa aaaagaaagt
     1801 agtaaaggat aagcattcgc ttacgttcaa aaatttgctg aacactgtag aaatgaagaa
     1861 caatctcaac aaggttgttg agaaaaagat cccaaaggag atcaccattg gaaataagtt
     1921 tgtgccagca tcggagaagc aacagaagat cagctacata tcggtatttc ctaagaatcc
     1981 gatgaatgga gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg
     2041 ccctatgcca cccaagcgca aaacgcttaa cgctgcggct gaggctaacg ctcatgccga
     2101 cggacacgcc gacggaaacg ccgacggaca cgtggccaat acggccgcgt cctcgaataa
     2161 tgcgaggttc gctgatctca ctaacatcga tactccgggt ctgggaccca caactacgac
     2221 cctgctcgtg gaaccagcac gctcaaagcg tcaacgagtg tcccgcgcat gcgaccagtg
     2281 ccgtgcagcc cgagagaaat gcgacggaat acagcctgcg tgtttcccgt gcgtttccca
     2341 gggaaggtcc tgcacttatc aggcttcgcc gaaaaagagg ggagttcaaa ccggttatat
     2401 tcgtacgctg gagctcgccc tcgcctggat gtttgaaaat gtcgcgcgtt ccgaagatgc
     2461 cttgcataac ctcctcgtcc gtgacgccgg acaaggatca gctctgctcg ttggtaaaga
     2521 ttcgccggct gccgagcgac tccatgcccg ttgggctact agccgtgtca ataagagcat
     2581 tacccgcctc ctccgtcagt tggagctccc tcctaccgcc acggctacgg cctcgataat
     2641 gccgcacgtg atggagcagc ctctcagtac cagcattaac cccgtcaacg accgcttcaa
     2701 cggtattccc aaccccactc cgtataactc cgatgcagct ctcgatgcta tcactcagac
     2761 caacgattat ggaagcgtaa atacacatgg tatcctctct acttacccgc caccggctac
     2821 gcaccttaat gaagcttccg tcgctctcgc tcccggtggc gccccccccc gaccgcctcc
     2881 tccgtatgtt gacagcacga ccaatcaccc gccgtaccac tcgaatctgg ttccaatggc
     2941 gaactttggt tactcgaccg ttgattacga tgccatggtt gacgatttgg ctagcattga
     3001 atacacggac gctgtggatg tcgacccaca gtttatgacc aatctgggat tcgttcctgg
     3061 atgtaacttc tccgacatta atacatacga acagtgatga cgtctatcga taccgtcgac
     3121 taaagccaaa tagaaaatta ttcagttcct ggcttaagtt tttaaaagtg atattattta
     3181 tttggttgta accaaccaaa agaatgtaaa taactaatac ataattatgt tagttttaag
     3241 ttagcaacaa attgatttta gctatattag ctacttggtt aataaataga atatatttat
     3301 ttaaagataa ttgcgttttt attgtcaggg agtgagtttg cttaaaaact cgtttagatc
     3361 caattcgagc tcgcccgggg atctaattca attagagact aattcaatta gagctaattc
     3421 aattaggatc caagcttatc gatttcgaac cctcgaccgc cggagtataa atagaggcgc
     3481 ttcgtctacg gagcgacaat tcaattcaaa caagcaaagt gaacacgtcg ctaagcgaaa
     3541 gctaagcaaa taaacaagcg cagctgaaca agctaaacaa tcggggtacc gctagagtcg
     3601 acggtaccgc gggcccggga tccaccggtc gccaccatgg cctcctccga gaacgtcatc
     3661 accgagttca tgcgcttcaa ggtgcgcatg gagggcaccg tgaacggcca cgagttcgag
     3721 atcgagggcg agggcgaggg ccgcccctac gagggccaca acaccgtgaa gctgaaggtg
     3781 accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc
     3841 tccaaggtgt acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc
     3901 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggc gaccgtgacc
     3961 caggactcct ccctgcagga cggctgcttc atctacaagg tgaagttcat cggcgtgaac
     4021 ttcccctccg acggccccgt gatgcagaag aagaccatgg gctgggaggc ctccaccgag
     4081 cgcctgtacc cccgcgacgg cgtgctgaag ggcgagaccc acaaggccct gaagctgaag
     4141 gacggcggcc actacctggt ggagttcaag tccatctaca tggccaagaa gcccgtgcag
     4201 ctgcccggct actactacgt ggacgccaag ctggacatca cctcccacaa cgaggactac
     4261 accatcgtgg agcagtacga gcgcaccgag ggccgccacc acctgttcct gtagcggccg
     4321 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     4381 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     4441 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4501 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaaat
     4561 gaatttgcaa ttaagggtca tttttaaatt acgtcacgct tgttgtacaa gttcatcgaa
     4621 acgtgacgac ccatacacaa ttttcgatgt ttccatgtgt gtgaaatttt gcgtgacgtg
     4681 atttatggac gtttcttaat tgaattatgg gcatcacaga taagtaaacc taagttgatg
     4741 aaattgataa ataaataaat tataatagtt tgttgaatca tagcttatgg gttactttcg
     4801 aagatttgaa cagaaaaacc ccgactaagt ccgttgagac tcgtatgttt gtctttattt
     4861 cgtgcctttt cgttttcagt tccttgtttt tcgctcgttt gctgtgtgtg tgttgggaca
     4921 caacagtagt caccttaaat actatttgca gtaattttct tgattttcca tccgtttgcc
     4981 gtaattctta cggttttatg aatttattac catcccttga cttgacttgg ctagacagta
     5041 caacatacgt gtgaacaaca catacaattt tactataggt ttgaaaagtt gtttgcccat
     5101 tagagggatt ggaaccacat ccaatctcta gaaaattctt attctgtttt ttggaggttg
     5161 aaatttagtg cactaacgaa aagtataaag gccatcaata aaaaaaagct tagagggaaa
     5221 cgattggctt cacgcaggta atcaagagaa gtacatttat tccacaatcc acaattgcta
     5281 ctcagtgtac atcaaactaa acgaatcatt acatgttatt gcattttcag cagcatctag
     5341 gagtgatgac ttcttctcta caaagggtat tattccatca cctacattcc gcatgcgcct
     5401 tgtgtggctt tactctttgt tgaggacaat cctatcgcgg tgtactatgc agttcttcgc
     5461 acgccgctgg gctcgcacta ggcagcataa ctgtctatgt gtgatgctgg agaagcgcat
     5521 cattgaatga ttttaatgtg gttgtcatcg tattgtatct gtgtacttca gttagtttgt
     5581 ttacgtcccg tatcgtttcg taccttttat cgtgtcgtaa taaagtatgt tttaaattgt
     5641 tacgtgcgcg tttaattccc tgatgtaggg acgagtttaa agaacaacca tcggggtatt
     5701 tccacggtac aaaagtggcg acgagaattc gcgagtgagg gtagtttttc cgtcgataaa
     5761 aactcaccca acatttcgtc acccgcgcgg aagaaaagaa agtggagtgc gatggctgac
     5821 ggtgatagtg atcgaaacgg gcaaaatggt gttgaccccc cgggagtgat aaatcggcag
     5881 caattaaacc agccgattat ccccgcgccg gcggctcacc agcagcagca gcagcaaatt
     5941 ttcgtccccg ttgtgactac acaccagcag cagcagcagc cattcgttcc ggtttcttca
     6001 gctccacaac agcagttctc aaccttccaa tcccaccaaa caccgcaagc ccaagtaagt
     6061 accgatatgc tcatgcaaat tatgcatatg atgcagcaat cgatgaccca gagtcagcag
     6121 cagcaccaac agcttatggg gcaaattatt gcgcagcagc agcagttcct aagtagcgtc
     6181 gcgtcgtcaa taaatgttca agtgccaccc aatccggaac aaatccttga ctccttggtc
     6241 tagagtcgac ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     6301 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6361 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6421 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6481 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6541 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6601 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6661 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6721 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6781 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6841 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6901 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6961 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7021 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7081 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     7141 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7201 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7261 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7321 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7381 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7441 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7501 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7561 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7621 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7681 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7741 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7801 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7861 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7921 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7981 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8041 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8101 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8161 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8221 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8281 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8341 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8401 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     8461 cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
