LOCUS       sequence-354642-        7872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for CRISPR-mediated knock-in line that expresses the 
            DNA-binding domain of QF by knocking-in a T2A-QF-DBD fragment 
            replacing the stop codon of the endogenous Aedes aegypti Orco gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7872)
  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 7872)
  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: ORCO-T2A-QF-DBD
FEATURES             Location/Qualifiers
     source          1..7872
                     /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             1623..1676
                     /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"
     intron          2499..2564
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2694..2714
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2986..3107
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      3386..3395
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3392..4069
                     /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(3572..3592)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3984..4007
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    4190..4311
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5651..5667)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5651..5667)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5664..5686)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5675..5691
                     /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(5699..5705,5706..5723,5724..5729))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5744..5765
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5882..5899)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6053..6641)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6133..6152)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6812..7603,7604..7672))
                     /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     7435..7454
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7673..7777)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7845..7863
                     /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 acccgggagt
      421 ttcgctttcc acgcgcggtg tgtctggaat cgttattaac gaaaaaaaat attctaattc
      481 gggatttaca attctctgtg aatatgaaaa catttcaacg agagaaacga aagttatagt
      541 gatttgaagg ttttcggcta ttctgtaaga tattctttca aataataggt gccaaattgg
      601 tggacatgtt ttgtaattaa cgatttcaga cacaccgtgc cacgtttggt tctctctagc
      661 aatttgattg tgcatttttt ttctctaaca gagttcatca gtgatggagg ctgcctactc
      721 gtgccactgg tatgacggtt ccgaggaagc caaaactttc gtgcaaatcg tttgccagca
      781 gtgtcagaaa gcgatgacca tatccggggc caagtttttc accgtttcac tggatctgtt
      841 cgcatcggtg agtgattttt ttttcgtttt cgattcggtt tctgggggga aattagtgtc
      901 ggatcgatcg ataaaagcga ttgcatgggc ataaccagaa aggagacatg agtcctcctg
      961 cctccacctc ccttttgaaa aaaagctggg tgaattaatt ttgtgtttgt attttcaaaa
     1021 tcgtctatat ctacaaaata attgcatcac gattgatatt tctgagaaat ttttgaagtt
     1081 actcaaacaa taattttcct aaaaaagatc ttatcagaaa ttatgagaag gttttatcaa
     1141 cagtccgcat ttttgaatca atataataag tttttctgaa aaaaaaaaaa tacagaaata
     1201 cggtgacgaa tttctgtgga atttttttga gattcaaata tttcaaaaac atattttatg
     1261 gtcgtctgca aaaaaccgtg agacgttttt gatttggtgg aatctatctt gaagatttct
     1321 tcgatgaact gtatgaaatt actatgtcga atcaattgaa tctgtagaaa aaatgtctca
     1381 tccggatttt tccaagcctt gtttctaatc gtccactgga ctccactttg gaaaagtgtg
     1441 aattttttaa gtcacttgcc cctccggcta ttctggctac gccaatgacc gattgtataa
     1501 gaaagccacg aaactgagtc aactataatt aatgattcgt cgttttcttt ccgttttcct
     1561 ttcccccaca ggttctggga gcggtcgtca cctacttcat ggtgctcgtg cagttgaaag
     1621 gagagggcag aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc ggccctatgc
     1681 tggagatccg cgccgccttc ctgcgccagc gcaacaccgc cctgcgcacc gaggtggccg
     1741 agctggagca ggaggtgcag cgcctggaga acgaggtgag ccagtacgag acccgctacg
     1801 gccccctggg cggcggcaag gctagcggag gaggtggtgg aggtggaggt ggaggtgacg
     1861 tcatgccacc caagcgcaaa acgcttaacg ctgcggctga ggctaacgct catgccgacg
     1921 gacacgccga cggaaacgcc gacggacacg tggccaatac ggccgcgtcc tcgaataatg
     1981 cgaggttcgc tgatctcact aacatcgata ctccgggtct gggacccaca actacgaccc
     2041 tgctcgtgga accagcacgc tcaaagcgtc aacgagtgtc ccgcgcatgc gaccagtgcc
     2101 gtgcagcccg agagaaatgc gacggaatac agcctgcgtg tttcccgtgc gtttcccagg
     2161 gaaggtcctg cacttatcag gcttcgccga aaaagagggg agttcaaacc ggttatattc
     2221 gtacgctgga gctcgccctc gcctggatgt ttgaaaatgt cgcgcgttcc gaagatgcct
     2281 tgcataacct cctcgtccgt gacgccggac aaggatcagc tctgctcgtt ggtaaagatt
     2341 cgccggctgc cgagcgactc catgcccgtt gggctactag ccgtgtcaat aagagcatta
     2401 cccgcctcct ctaaggccgg ccgatctttg tgaaggaacc ttacttctgt ggtgtgacat
     2461 aattggacaa actacctaca gagatttaaa gctctaaggt aaatataaaa tttttaagtg
     2521 tataatgtgt taaactactg attctaattg tttgtgtatt ttagattcca acctatggaa
     2581 ctgatgaatg ggagcagtgg tggaatgcct ttaatgagga aaacctgttt tgctcagaag
     2641 aaatgccatc tagtgatgat gaggctactg ctgactctca acattctact cctccaaaaa
     2701 agaagagaaa ggtagaagac cccaaggact ttccttcaga attgctaagt tttttgagtc
     2761 atgctgtgtt tagtaataga actcttgctt gctttgctat ttacaccaca aaggaaaaag
     2821 ctgcactgct atacaagaaa attatggaaa aatatttgat gtatagtgcc ttgactagag
     2881 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2941 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3001 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3061 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgaatt
     3121 cgagctcgcc cggggatcta attcaattag agactaattc aattagagct aattcaatta
     3181 ggatccaagc ttatcgattt cgaaccctcg accgccggag tataaataga ggcgcttcgt
     3241 ctacggagcg acaattcaat tcaaacaagc aaagtgaaca cgtcgctaag cgaaagctaa
     3301 gcaaataaac aagcgcagct gaacaagcta aacaatcggg gtaccgctag agtcgacggt
     3361 accgcgggcc cgggatccac cggtcgccac catggcctcc tccgagaacg tcatcaccga
     3421 gttcatgcgc ttcaaggtgc gcatggaggg caccgtgaac ggccacgagt tcgagatcga
     3481 gggcgagggc gagggccgcc cctacgaggg ccacaacacc gtgaagctga aggtgaccaa
     3541 gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttccagt acggctccaa
     3601 ggtgtacgtg aagcaccccg ccgacatccc cgactacaag aagctgtcct tccccgaggg
     3661 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggcgaccg tgacccagga
     3721 ctcctccctg caggacggct gcttcatcta caaggtgaag ttcatcggcg tgaacttccc
     3781 ctccgacggc cccgtgatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct
     3841 gtacccccgc gacggcgtgc tgaagggcga gacccacaag gccctgaagc tgaaggacgg
     3901 cggccactac ctggtggagt tcaagtccat ctacatggcc aagaagcccg tgcagctgcc
     3961 cggctactac tacgtggacg ccaagctgga catcacctcc cacaacgagg actacaccat
     4021 cgtggagcag tacgagcgca ccgagggccg ccaccacctg ttcctgtagc ggccgcgact
     4081 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     4141 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     4201 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4261 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt ataattcgtt
     4321 aattatggac aagctcaaga ctatccgata ctatttttga tactaatgca actgaacggc
     4381 gatacatgac atgaaatcgt ttttatatgt agatttataa ggaagaaaaa aattatcagt
     4441 acaagaacaa ttagactaga ccctccgaca ctgagtagcg gaggctaatg atgttgatat
     4501 attaaatatt ttagtaagaa tttttgatgt agctgaaata gacgctcaag atgaatgcaa
     4561 tgaactctcg agaacttatt cataacagtg agagaatgcc ctgattgttc actgtaatta
     4621 gttttccaca cgatttactt ggtttatgaa aagcccttgt agattgagaa gtcattagtc
     4681 aagatatgtc actcccgtgc aaaagtttgg gttcaccccc tcaaacatat acaaaagtgt
     4741 tctgtccata tctctgtgat tacacgtcaa atttaaactc tttaagccgc attcgaaagg
     4801 caaagagtta ttcttacttc gtatgtattt ttccaaaaac atttttttaa attttgtata
     4861 ctaaattttt attaaagttg tgacattttt caaaaaacac ccttaaaaat catatctaat
     4921 ttcctcagca ttgggttgac caaaatttca aatcaaagtg tcattagaat cgtaatctta
     4981 tattccttga agagacacca cgaaatttcg gcggacaaat ctggaaagta ttcaaaatca
     5041 atgaaacagt cagtcaagtc atcgtgcaaa agtttgggtt catctctcag tatgatgcaa
     5101 atcgtgcaaa agtttgggtt cacctgatcc gcacgcacat aatttttgtc aatatctcgg
     5161 ccatttttca accgatttta atagtttaaa gcttttttga gcgcaaataa tggcgtgtac
     5221 atgattggtc tgagatttca cagatttaag taagttcagg tgaacccaaa tttttgcacg
     5281 atacaccata cactgagggg tgaacccaaa cttttgcacg atgacttgac tgactgtttt
     5341 attgattttg aatacttctc agatttttcc gccaaaattt aatgagtgct cttcaaagta
     5401 tataagatta cgattctaat gacaccttgt tttaaaattt tggtcaattg aatgctgagg
     5461 aaattagata tgatttttca gtgtgttttt tgaaaaatgt cccaacttta agtacaaatt
     5521 tagtatacaa agttcaaaga atgtttttgg aaaaatacat acgaagtaag aataactctt
     5581 tgcctttcga atgcggctta gagagttttc tagagtcgac ctgcaggcat gcaagcttgg
     5641 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5701 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5761 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5821 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5881 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5941 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6001 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6061 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6121 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6181 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6241 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6301 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6361 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6421 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6481 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6541 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6601 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6661 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6721 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6781 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6841 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6901 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6961 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7021 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7081 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7141 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7201 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7261 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7321 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7381 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7441 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7501 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7561 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7621 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7681 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7741 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7801 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     7861 ggccctttcg tc
//
