LOCUS       sequence-372762-        7441 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 Gr3 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7441)
  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 7441)
  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: GR3-T2A-QF2
FEATURES             Location/Qualifiers
     source          1..7441
                     /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             2247..2300
                     /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      3887..3896
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3893..4570
                     /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(4073..4093)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     4485..4508
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    4691..4812
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4728..4747)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4782..4801
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5220..5236)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5220..5236)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5233..5255)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5244..5260
                     /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(5268..5274,5275..5292,5293..5298))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5313..5334
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5451..5468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5622..6210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5702..5721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6381..7172,7173..7241))
                     /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     7004..7023
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7242..7346)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7414..7432
                     /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 acccggcttt
      421 atgatggggc cgtccgctgt ttgcgcgttt gggtagtgaa ataattcctc ttgatgatgg
      481 cgctcatgtg acgatgatgc gttaaatatt catcggaagc ggtggcgaag gagtcggaac
      541 tttgcccttg ttgtttttct tacgatggtc agggtcgtcg cttttgcatg ctaacatcgt
      601 aaattttaaa gtgacactgg gaacttttcc aattactaaa cctttttgtc gctaagttga
      661 cgaaaaacgt gatccagtat gctagagttc tcgactgagt aaggtgttct agagaattga
      721 caatgaatct caaccaagac cccattcagt acatcaattt gaataacaat gctcgaacgg
      781 tttttctgga cgtgaaacca atttacaacg aagagaagcg taaagtttct aatggattta
      841 acaatcgcat tggatttccg ccaatctctt cgaggagagt gttcggtttg gaaagtgaat
      901 ttaacacgcg atcggtgagt caaactgcag aagcttcctt ccgtccagta acgatcttac
      961 catatacaat cgcaggatat agtttacggc accacgaagc caatctacaa cgttctacgg
     1021 atgctgggag tgtttccttt ctcgagacct tcacccggag tgacattatt cgcctgtgcg
     1081 agccctgcga tggcatattg cggtgtgctt ttcgtgacgc ttatggtaat ttccgtatga
     1141 ctaagtttgg gtaatactcc gcgcggctaa ccttgccgat catatatatt cgtaggctta
     1201 cgtcatctac ataaccatcc tccgggtcca tattgtccgc acactggaag gccgcttcga
     1261 agaggccgtc atcgcttatc tcttcatcgt taacatcctg ccggttctga tcattccgtt
     1321 gatgtggtac gaaactcgta aggtctccag tttgctcaat cagtgggtcg acttcgaggc
     1381 gatctatcgt aaaactgcgg gcagagaact ggagctgtca ttcagaacca aagctctact
     1441 gattgctatt ttactgccgg ttctatcttg cctggcagtc atcataaccc acgtgacaat
     1501 ggttgagttc cagttggttc aggtcatccc gtattgcatt ttggacacgc taacctacat
     1561 gatgggaggt tattggtata tgacctgcga aactctcagt ataactgcta acattctggc
     1621 ggaggatttc caaagggctt tgagacacgt tggaccagca gctatggtat ccgaataccg
     1681 ctcgctatgg ctgcgattga gcaagcttgc tcgagaaacg ggatcatcca cgtgttacac
     1741 cttcacattt ctgtgtctat acctcttctt catcatcact ctttcgatct acggcctgat
     1801 gtcgcaaatt tccgaaggtt tcggcatcaa agacatcggc ctggcagtga ccgccttctg
     1861 cagcgttggg ttactcttct tcatatgcga cgaagctcac tacgcgtcgt tcaacgttcg
     1921 gaccaaattc cagaagaagt tgttgatggc agagctcagt tggatgaact cggatgcaca
     1981 aaccgaaatc aacatgtttt tgagggcaac cgagatgaat ccttcgagca tcaacttggg
     2041 cgggtttttc gacgtgaacc ggacgctgtt caaatcggtg agtttagtcc tgcaagattg
     2101 aataaccctg ttttaaaata attccattcc agcttttggc aacgatggtg acctatttgg
     2161 tggtgttgct acagttccaa atcagcatac cagacgactc tagcatgctt gttatgcaca
     2221 acatgaccgg ttcatatcgc gagggagagg gcagaggaag tcttctaaca tgcggtgacg
     2281 tggaggagaa tcccggccct atgccaccca agcgcaaaac gcttaacgct gcggctgagg
     2341 ctaacgctca tgccgacgga cacgccgacg gaaacgccga cggacacgtg gccaatacgg
     2401 ccgcgtcctc gaataatgcg aggttcgctg atctcactaa catcgatact ccgggtctgg
     2461 gacccacaac tacgaccctg ctcgtggaac cagcacgctc aaagcgtcaa cgagtgtccc
     2521 gcgcatgcga ccagtgccgt gcagcccgag agaaatgcga cggaatacag cctgcgtgtt
     2581 tcccgtgcgt ttcccaggga aggtcctgca cttatcaggc ttcgccgaaa aagaggggag
     2641 ttcaaaccgg ttatattcgt acgctggagc tcgccctcgc ctggatgttt gaaaatgtcg
     2701 cgcgttccga agatgccttg cataacctcc tcgtccgtga cgccggacaa ggatcagctc
     2761 tgctcgttgg taaagattcg ccggctgccg agcgactcca tgcccgttgg gctactagcc
     2821 gtgtcaataa gagcattacc cgcctcctcc gtcagttgga gctccctcct accgccacgg
     2881 ctacggcctc gataatgccg cacgtgatgg agcagcctct cagtaccagc attaaccccg
     2941 tcaacgaccg cttcaacggt attcccaacc ccactccgta taactccgat gcagctctcg
     3001 atgctatcac tcagaccaac gattatggaa gcgtaaatac acatggtatc ctctctactt
     3061 acccgccacc ggctacgcac cttaatgaag cttccgtcgc tctcgctccc ggtggcgccc
     3121 ccccccgacc gcctcctccg tatgttgaca gcacgaccaa tcacccgccg taccactcga
     3181 atctggttcc aatggcgaac tttggttact cgaccgttga ttacgatgcc atggttgacg
     3241 atttggctag cattgaatac acggacgctg tggatgtcga cccacagttt atgaccaatc
     3301 tgggattcgt tcctggatgt aacttctccg acattaatac atacgaacag tgatgacgtc
     3361 tatcgatacc gtcgactaaa gccaaataga aaattattca gttcctggct taagttttta
     3421 aaagtgatat tatttatttg gttgtaacca accaaaagaa tgtaaataac taatacataa
     3481 ttatgttagt tttaagttag caacaaattg attttagcta tattagctac ttggttaata
     3541 aatagaatat atttatttaa agataattgc gtttttattg tcagggagtg agtttgctta
     3601 aaaactcgtt tagatccaat tcgagctcgc ccggggatct aattcaatta gagactaatt
     3661 caattagagc taattcaatt aggatccaag cttatcgatt tcgaaccctc gaccgccgga
     3721 gtataaatag aggcgcttcg tctacggagc gacaattcaa ttcaaacaag caaagtgaac
     3781 acgtcgctaa gcgaaagcta agcaaataaa caagcgcagc tgaacaagct aaacaatcgg
     3841 ggtaccgcta gagtcgacgg taccgcgggc ccgggatcca ccggtcgcca ccatggcctc
     3901 ctccgagaac gtcatcaccg agttcatgcg cttcaaggtg cgcatggagg gcaccgtgaa
     3961 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gccacaacac
     4021 cgtgaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc
     4081 ccagttccag tacggctcca aggtgtacgt gaagcacccc gccgacatcc ccgactacaa
     4141 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     4201 cgtggcgacc gtgacccagg actcctccct gcaggacggc tgcttcatct acaaggtgaa
     4261 gttcatcggc gtgaacttcc cctccgacgg ccccgtgatg cagaagaaga ccatgggctg
     4321 ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agacccacaa
     4381 ggccctgaag ctgaaggacg gcggccacta cctggtggag ttcaagtcca tctacatggc
     4441 caagaagccc gtgcagctgc ccggctacta ctacgtggac gccaagctgg acatcacctc
     4501 ccacaacgag gactacacca tcgtggagca gtacgagcgc accgagggcc gccaccacct
     4561 gttcctgtag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     4621 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     4681 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4741 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4801 caatgtatct tatagatcaa tatcctgatg attgagaaaa ggaacttctc tcgaagtaat
     4861 aaatgtttca ttagtgttgc ttattattaa ctttaattta atttttatta tccctgaagg
     4921 acctcgcatc ttattcctgc gggtgccttt tgagaatgcg tatggggttt cttttttgac
     4981 ttatttcaga gttaaaacct ccagctacta ctaaatttcc tcatgaattt cttttagaat
     5041 ggattgggtt agaagcaaaa ggttgtaagt gacaacttta atgttttctc agtttttttt
     5101 taattccgta caaattgtat gtgagcccag aaataagcca ctttactttt gatggcgcta
     5161 cgtccttcaa gacatcctct agagtcgacc tgcaggcatg caagcttggc gtaatcatgg
     5221 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5281 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5341 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5401 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5461 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5521 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5581 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5641 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5701 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5761 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5821 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5881 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5941 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6001 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6061 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6121 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6181 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6241 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6301 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6361 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6421 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6481 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6541 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6601 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6661 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6721 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6781 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6841 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6901 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6961 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7021 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7081 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7141 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7201 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7261 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7321 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     7381 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     7441 c
//
