LOCUS       sequence-354669-        8624 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 Ir76b gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8624)
  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 8624)
  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: IR76b-T2A-QF2
FEATURES             Location/Qualifiers
     source          1..8624
                     /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             1630..1683
                     /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      3270..3279
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3276..3953
                     /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(3456..3476)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3868..3891
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    4074..4195
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4111..4130)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4165..4184
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6403..6419)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6403..6419)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6416..6438)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6427..6443
                     /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(6451..6457,6458..6475,6476..6481))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6496..6517
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6634..6651)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6805..7393)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6885..6904)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7564..8355,8356..8424))
                     /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     8187..8206
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8425..8529)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8597..8615
                     /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 acccggtttc
      421 tactgttacg aatagaaaag cggggtgtct gcctttcttt attgaaatca aaaatcaaca
      481 actcgccgtc tatagtggtt acacgtgttc gccattggcg tagaccaaaa ataaatagaa
      541 ttttcgattg ggtctatcac acacccatga gcagttagac aaaaaaactt attggtgaag
      601 gtataaatca atgcacgtca gcctagacaa cctttccgaa ccttatcccc tgcccccttc
      661 ctaacagttg ctgttttcat agagcgacga gaccctgaac tcgttgagtt ttttggtgga
      721 caaacacttg gtggagttca caaccaacct gaacgatagc gacatcctga tgacaacggt
      781 gtccaagaaa aactacgtct acgtacggga tcgaccggcc atcgaacatc tggtttacaa
      841 agactacctc gttcggcgca aggtgaaccc catcaacgaa cgtagccatt gtccgtttgc
      901 aacggctaca acacctttcc tgaaacgggt tcgcgccttt gcctatccca ataccacacg
      961 ctggaacacg ctatttgatc cacagtaagt tctactaaaa gtataacaaa gacgccatgt
     1021 ataaaccatt tcttcgttct acgcagacta ctcaacatgg tcgaaggagg cttggtgaaa
     1081 ttcaagcttc tggagaatct acccaaggcc gagatctgtc cgcagaactt aggaggcact
     1141 gagcgtcagc tgaagaatcg ggacttggtc atgacatact tggtgatgtt gaccggcttt
     1201 gccacggcaa ttgctgtatt cgtttcggag gtaactacat ctgcttgatc acagctctag
     1261 taggccgtta acaatattct tcttgttgtc ttcctataga tgttcttccg catcctgaac
     1321 gagcgcaaac tgaacaaaga acagtcggaa gctgcccgga accgtggcac cgatggagtt
     1381 acctatgtga atgcaaaggc catgggttat ggcgattcac cgccgccgcc gtatgcggag
     1441 attttcagcc gccatgctag cggtggaagc atgtttggaa gcagtattga cagtggtagc
     1501 ggtgcatcac ggaggatgat caacggcagg gattacatgg tcgtcaagga gcgaaatggg
     1561 atcggttcgc agctcatacc aattcgggcg ccgtcagctg caatttttca gtacacttat
     1621 acgaatggag agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc
     1681 cctatgccac ccaagcgcaa aacgcttaac gctgcggctg aggctaacgc tcatgccgac
     1741 ggacacgccg acggaaacgc cgacggacac gtggccaata cggccgcgtc ctcgaataat
     1801 gcgaggttcg ctgatctcac taacatcgat actccgggtc tgggacccac aactacgacc
     1861 ctgctcgtgg aaccagcacg ctcaaagcgt caacgagtgt cccgcgcatg cgaccagtgc
     1921 cgtgcagccc gagagaaatg cgacggaata cagcctgcgt gtttcccgtg cgtttcccag
     1981 ggaaggtcct gcacttatca ggcttcgccg aaaaagaggg gagttcaaac cggttatatt
     2041 cgtacgctgg agctcgccct cgcctggatg tttgaaaatg tcgcgcgttc cgaagatgcc
     2101 ttgcataacc tcctcgtccg tgacgccgga caaggatcag ctctgctcgt tggtaaagat
     2161 tcgccggctg ccgagcgact ccatgcccgt tgggctacta gccgtgtcaa taagagcatt
     2221 acccgcctcc tccgtcagtt ggagctccct cctaccgcca cggctacggc ctcgataatg
     2281 ccgcacgtga tggagcagcc tctcagtacc agcattaacc ccgtcaacga ccgcttcaac
     2341 ggtattccca accccactcc gtataactcc gatgcagctc tcgatgctat cactcagacc
     2401 aacgattatg gaagcgtaaa tacacatggt atcctctcta cttacccgcc accggctacg
     2461 caccttaatg aagcttccgt cgctctcgct cccggtggcg cccccccccg accgcctcct
     2521 ccgtatgttg acagcacgac caatcacccg ccgtaccact cgaatctggt tccaatggcg
     2581 aactttggtt actcgaccgt tgattacgat gccatggttg acgatttggc tagcattgaa
     2641 tacacggacg ctgtggatgt cgacccacag tttatgacca atctgggatt cgttcctgga
     2701 tgtaacttct ccgacattaa tacatacgaa cagtgatgac gtctatcgat accgtcgact
     2761 aaagccaaat agaaaattat tcagttcctg gcttaagttt ttaaaagtga tattatttat
     2821 ttggttgtaa ccaaccaaaa gaatgtaaat aactaataca taattatgtt agttttaagt
     2881 tagcaacaaa ttgattttag ctatattagc tacttggtta ataaatagaa tatatttatt
     2941 taaagataat tgcgttttta ttgtcaggga gtgagtttgc ttaaaaactc gtttagatcc
     3001 aattcgagct cgcccgggga tctaattcaa ttagagacta attcaattag agctaattca
     3061 attaggatcc aagcttatcg atttcgaacc ctcgaccgcc ggagtataaa tagaggcgct
     3121 tcgtctacgg agcgacaatt caattcaaac aagcaaagtg aacacgtcgc taagcgaaag
     3181 ctaagcaaat aaacaagcgc agctgaacaa gctaaacaat cggggtaccg ctagagtcga
     3241 cggtaccgcg ggcccgggat ccaccggtcg ccaccatggc ctcctccgag aacgtcatca
     3301 ccgagttcat gcgcttcaag gtgcgcatgg agggcaccgt gaacggccac gagttcgaga
     3361 tcgagggcga gggcgagggc cgcccctacg agggccacaa caccgtgaag ctgaaggtga
     3421 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc cagtacggct
     3481 ccaaggtgta cgtgaagcac cccgccgaca tccccgacta caagaagctg tccttccccg
     3541 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggcg accgtgaccc
     3601 aggactcctc cctgcaggac ggctgcttca tctacaaggt gaagttcatc ggcgtgaact
     3661 tcccctccga cggccccgtg atgcagaaga agaccatggg ctgggaggcc tccaccgagc
     3721 gcctgtaccc ccgcgacggc gtgctgaagg gcgagaccca caaggccctg aagctgaagg
     3781 acggcggcca ctacctggtg gagttcaagt ccatctacat ggccaagaag cccgtgcagc
     3841 tgcccggcta ctactacgtg gacgccaagc tggacatcac ctcccacaac gaggactaca
     3901 ccatcgtgga gcagtacgag cgcaccgagg gccgccacca cctgttcctg tagcggccgc
     3961 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     4021 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     4081 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4141 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaggtga
     4201 ctttggaaag tagccccact tttttggtat aggtattgtt cagcatcaat cataaatagt
     4261 taaccttcaa gagtcgctgt tacatgactc gatatcatgt aacggttgtc gatggaaagt
     4321 atgcaaagtg ctgtttgtct gttttagagt taaaatttca tagttactat gatggtcctc
     4381 tctcaaataa gtttccaaaa catttatata gatctagagt tttagtaaat gtagattttc
     4441 gttatctatc cagaccagta ccggagattt cattttaaat ctttttattt ggcatagcac
     4501 actaacgaaa aattgcagtg acttagggac cattgtgcat acgaattcaa atacagtgaa
     4561 ctctctttca atcggtattg aagggaccat cgagttaggg aggtatcgag atgcagaata
     4621 catatttttc agttttcagt ttcaatttta gctttaaaac attaatttta aaaaagcgtt
     4681 cgaaataata gttttaacgt taaaaattaa agaatactaa gcaacacttt ttgaacaatc
     4741 ttgaaaaact tcagattgtt tcatccttaa atgggttgtc aaccttaatt ttacaatgat
     4801 catagttaaa atttttgaaa tctattaaca ttcggagggt gtatgaaact tttcatgtaa
     4861 acttcgagtt agatagatat aaagttgcag gaaactagaa cagattacat agatgtgctt
     4921 attctgcgcg acgtgtgagt cgagacgagt cgctctcacc gcgagtgacg tgagacgact
     4981 aatgttaata aaatgggagc gagtcgacaa ttgtcacctc attcgactca tctcacctca
     5041 cacgccgcgc agaataaaca cgagagagat caaagtatgc tagattgaag ggactgcaca
     5101 ttccatcgta ttagggaaaa tatcgtgtta caaaatgtcg agtaagggag agctgacttt
     5161 attgccaata actgcaaatt agtcacattt gacatttttt accatttcga gttaattgtg
     5221 cttattctgc gcggcgtgtg agtcgagacg agtcgctctc acctcgggtg acgtgagacg
     5281 actaatgtta atcaaatggg agcgagtcga caattgtcac ctcattcgac tcgtgtcacc
     5341 tcacacgccg cgcagaatag gcacgaatac catgcagggt catcaaatga taaacacttt
     5401 cgatccactt actgagctct gtgagattgt tctagaaaac tcgaaaaaaa aaataccaaa
     5461 atgttttgtc acattggcaa tggtaaccgc ataacagtca cattgagatt atatatgagc
     5521 ctcgtaatgt aatgaaataa tgaaatttct atcagatttt tttctggcaa tatcgcatat
     5581 atgagctgta cgaaatttca gcatcaaata agcacttttg tgtttttggt aattttaaga
     5641 tattttagta tcttcccata ctgcacactc agttgagctc ggctaatttt cattcttttg
     5701 ccgagatccg cacagctgag caactgaata aagtgtgtgt tatggggctt tgcacgaaaa
     5761 tccatccctc tctttcacta tgtgctttca ctcttccatg aaattagaaa acatcaagac
     5821 cagtaaacgt caaaatccca tacaaaatca gaaacttcat aaaatgcaca cttggtactc
     5881 catttactca atgcctactt ttgtcgaatg cttcaaatat gcaagggatc gatcaggtat
     5941 ctgaggaatt ttttttcaag ctcacgatca aaagctacac acgtgctcat atttgtgtcc
     6001 aagttttttt ttcgaataca ggccttattc gaagctcaat tgcataatac gctctaagga
     6061 gcagactttt taattgtcct aatatagtgg aatgatgcca aggtactaga cacaacttaa
     6121 gtttcgtagg atacgaatat gctaggatgg ataggacaat gatgtgtcaa aaagattcac
     6181 tggagctctt ctgagtacat cttagcaact ttaggtaaga gacaatccac aagccacgtg
     6241 gtcatatttt tgggttgtag tcttttgttc atgcagaaca tttaaaattt gtgtgtccac
     6301 cctaagtgag cacgtggttt atggacgacc gcgaaggtaa cttgtgtagt ttcggcaacc
     6361 tctagagtcg acctgcaggc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt
     6421 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6481 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6541 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6601 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6661 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6721 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6781 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6841 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6901 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6961 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7021 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7081 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7141 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7201 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     7261 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7321 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7381 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7441 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7501 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7561 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7621 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7681 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7741 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7801 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7861 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7921 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7981 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8041 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8101 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8161 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8221 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8281 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8341 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8401 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8461 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8521 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     8581 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc
//
