LOCUS       sequence-215869-        7404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Flpo recombinase-dependent GtACR2-KA2N, which targets 
            GtACR2 to the somatodendritic compartment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7404)
  AUTHORS   Messier JE, Chen H, Cai ZL, Xue M
  TITLE     Targeting light-gated chloride channels to neuronal somatodendritic 
            domain reduces their excitatory effect in the axon.
  JOURNAL   Elife. 2018 Aug 9;7. pii: 38506. doi: 10.7554/eLife.38506.
  PUBMED    30091701
REFERENCE   2  (bases 1 to 7404)
  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"; journalName: "Elife."; 
            date: "2018-08-9"; volume: "7. pii: 38506. doi"; pages: " 
            10.7554/eLife.38506"
COMMENT     Sequence Label: pAAV-EF1-alpha-FRT-FLEX-GtACR2-KA2N-EGFP
FEATURES             Location/Qualifiers
     source          1..7404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1420
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1411
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1368..1388
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1511..1544
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(join(1557..2270,2271..2273))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(1560..2270,2271..2273))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1602..1623)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1951..1970
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2210..2231
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2292..2741)
                     /codon_start=1
                     /product="first 150 residues of the mouse kainate receptor 
                     subunit 2"
                     /label=KA2
                     /note="mediates targeting to the neuronal soma (Shemesh et 
                     al., 2017)"
                     /translation="MPAELLLLLIVAFANPSCQVLSSLRMAAILDDQTVCGRGERLALA
                     LAREQINGIIEVPAKARVEVDIFELQRDSQYETTDTMCQILPKGVVSVLGPSSSPASAS
                     TVSHICGEKEIPHIKVGPEETPRLQYLRFASVSLYPSNEDVSLAVS"
     protein_bind    complement(3771..3804)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    3823..4411
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3876..3896)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4294..4305)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4444..4555
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4667..4807
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4667..4796
                     /label=AAV2 ITR
     rep_origin      4882..5337
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4969..4988)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5179..5200
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5354..5373)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5473..5495
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5533..5551)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5619..5723
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5724..5792,5793..6584)
                     /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     complement(5942..5961)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6755..7343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7244..7263
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg
      601 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg
      661 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
      721 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
      781 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc
      841 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
      901 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
      961 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
     1021 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
     1081 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
     1141 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
     1201 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     1261 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     1321 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     1381 agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggtaccgaag ttcctattct
     1441 atcagaagta taggaacttc gcagaatggt agctggattg tagctgctat tagcaatatg
     1501 aaacctctta gaagttccta ttctctagaa agtataggaa cttcgaattc aagcttttac
     1561 ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc
     1621 atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct caggtagtgg
     1681 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg
     1741 agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc
     1801 ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc
     1861 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg
     1921 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag
     1981 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc
     2041 atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt cacgagggtg
     2101 ggccagggca cgggcagctt gccggtggtg caaatgaact tcagggtcag cttgccgtag
     2161 gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc
     2221 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct cacagtacct
     2281 ccgctagctc cgctgactgc cagtgacaca tcctcgtttg atggatacag gctcactgag
     2341 gcgaagcgca ggtactgcag cctgggtgtt tcctctggcc cgaccttgat atgggggatt
     2401 tccttctcgc cacaaatgtg agacacggta cttgcagaag caggagagga gcttggtccc
     2461 aggacggaga ccactccttt tggcaggatc tggcacatgg tgtctgtggt ctcgtactgg
     2521 ctatctctct gcagttcgaa aatgtccacc tcgacccggg cctttgcagg cacttcaatg
     2581 atgccattga tctgctccct agccagtgcc agtgccagcc tctctcctct tccgcacacg
     2641 gtctgatcgt ccagaatagc ggccatcctc agagaagaca gcacctggca agaagggtta
     2701 gcaaaagcga caatcagcag cagcagcagt tcggcgggca tggtacctcc acttccaccg
     2761 gttccacctg aacctcccct gccgaacatt ctgatgttga cctcgtcgtt ggggggggca
     2821 ttcttctcgt ggttcagtcc gaaggggtcg tcggcctcag gcttctcctg gccagggagg
     2881 caagccgggt cccactttcc ctcgagcact ccccaggcgg tggaccacag gatcacgccg
     2941 taggtgttct tgcacaggcc gtcggcgagc agatacagga tggcgctcac gttctcgtgg
     3001 atcacgcaca cgccggtggg ggaaatcagc cacacgatgg ggaacatggt ccaggcgatg
     3061 aagaagatca gcctcagcat cttgatcctc accaccacct tctgggccag aggggtgccc
     3121 acggactcga agccgtacag tcccacctgg aagatggtga acaccacgct gtattcgaag
     3181 accaggcagg tagcgccgaa gaagaagaac agccacttca tgtactcggc gggggagatg
     3241 gtagcggtaa tgcccatcac caccctgata atgctggcgg cctgggcgat ggggttcaga
     3301 gggatgctct tgtacttcac cagagccatg ttggagatct ggcccagcat aatggggcat
     3361 gtgcacagcc agctggccat cctggaccag ggaaagaacc tgccgtccct catctgcagt
     3421 gtgccattgc cggtgctggc cagggcgtat gtgatggact ccacgaaggg caggtacacg
     3481 ctctcccagg gcaccttggg ggccctgctg atcatgaaga tggtctggca tccgctcacc
     3541 acggcccaca ccagctgcag caggctcacg aaggtgctgt cgatcctgct catgttgtag
     3601 cactctgtgt gggtgctggc ccactcgccg tacacgacct gggaggccat ggatcccggc
     3661 tagctacgta ggcgcgcctt aattaagaag ttcctatact tctgatagaa taggaacttc
     3721 ttgccttaac ccagaaatta tcactgttat tctttagaat ggtgcaaaga gaagttccta
     3781 tactttctag agaataggaa cttccagatc aagcttatcg ataatcaacc tctggattac
     3841 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3901 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3961 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     4021 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     4081 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     4141 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     4201 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg
     4261 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     4321 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     4381 agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct
     4441 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     4501 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     4561 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     4621 attgggaaga caatagcagg catgcacgtg cggaccgagc ggccgcagga acccctagtg
     4681 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     4741 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     4801 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4861 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4921 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4981 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     5041 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     5101 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5161 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5221 cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5281 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     5341 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     5401 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     5461 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     5521 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     5581 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5641 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5701 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5761 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5821 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5881 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5941 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     6001 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6061 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6121 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     6181 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6241 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     6301 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6361 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6421 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6481 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6541 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6601 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6661 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6721 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6781 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6841 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6901 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6961 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7021 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7081 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     7141 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     7201 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     7261 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7321 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7381 tttgctggcc ttttgctcac atgt
//
