LOCUS       sequence-215862-        7269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Flpo recombinase-dependent GtACR2-Kv4.2LL, which targets 
            GtACR2 to the somatodendritic compartment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7269)
  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 7269)
  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-EYFP-Kv4.2LL
FEATURES             Location/Qualifiers
     source          1..7269
                     /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..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1510..1543
                     /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(1620..2330,2331..2333))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1662..1683)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2011..2030
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2270..2291
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(3336..3369)
                     /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    3396..3984
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3449..3469)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3867..3878)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4016..4492
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4215..4234)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4532..4672
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4532..4661
                     /label=AAV2 ITR
     rep_origin      4747..5202
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4834..4853)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5044..5065
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5219..5238)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5338..5360
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5398..5416)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5484..5588
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5589..5657,5658..6449)
                     /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(5807..5826)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6620..7208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7109..7128
                     /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 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccgaagt tcctattcta
     1441 tcagaagtat aggaacttcg cagaatggta gctggattgt agctgctatt agcaatatga
     1501 aacctcttag aagttcctat tctctagaaa gtataggaac ttcgaattct tacttgtaca
     1561 gtttaaactc acgtggtttt ctccaggcag tgaagcaggt ggtggtgttg tgtctcgaac
     1621 ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc
     1681 atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actggtagct caggtagtgg
     1741 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg
     1801 agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc
     1861 ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc
     1921 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg
     1981 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag
     2041 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc
     2101 atgtggtcgg ggtagcgggc gaagcactgc aggccgtagc cgaaggtggt cacgagggtg
     2161 ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag
     2221 gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc
     2281 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct cacggcggcc
     2341 gccctgccga acattctgat gttgacctcg tcgttggggg gggcattctt ctcgtggttc
     2401 agtccgaagg ggtcgtcggc ctcaggcttc tcctggccag ggaggcaagc cgggtcccac
     2461 tttccctcga gcactcccca ggcggtggac cacaggatca cgccgtaggt gttcttgcac
     2521 aggccgtcgg cgagcagata caggatggcg ctcacgttct cgtggatcac gcacacgccg
     2581 gtgggggaaa tcagccacac gatggggaac atggtccagg cgatgaagaa gatcagcctc
     2641 agcatcttga tcctcaccac caccttctgg gccagagggg tgcccacgga ctcgaagccg
     2701 tacagtccca cctggaagat ggtgaacacc acgctgtatt cgaagaccag gcaggtagcg
     2761 ccgaagaaga agaacagcca cttcatgtac tcggcggggg agatggtagc ggtaatgccc
     2821 atcaccaccc tgataatgct ggcggcctgg gcgatggggt tcagagggat gctcttgtac
     2881 ttcaccagag ccatgttgga gatctggccc agcataatgg ggcatgtgca cagccagctg
     2941 gccatcctgg accagggaaa gaacctgccg tccctcatct gcagtgtgcc attgccggtg
     3001 ctggccaggg cgtatgtgat ggactccacg aagggcaggt acacgctctc ccagggcacc
     3061 ttgggggccc tgctgatcat gaagatggtc tggcatccgc tcaccacggc ccacaccagc
     3121 tgcagcaggc tcacgaaggt gctgtcgatc ctgctcatgt tgtagcactc tgtgtgggtg
     3181 ctggcccact cgccgtacac gacctgggag gccatggatc ccggctagct acgtaggcgc
     3241 gccttaatta agaagttcct atacttctga tagaatagga acttcttgcc ttaacccaga
     3301 aattatcact gttattcttt agaatggtgc aaagagaagt tcctatactt tctagagaat
     3361 aggaacttcc agaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     3421 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3481 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3541 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3601 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3661 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3721 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3781 tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc
     3841 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3901 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3961 tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg
     4021 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     4081 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     4141 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     4201 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     4261 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     4321 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     4381 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     4441 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
     4501 gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     4561 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     4621 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     4681 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     4741 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4801 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4861 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4921 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     4981 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5041 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg
     5101 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     5161 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     5221 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     5281 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     5341 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     5401 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     5461 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5521 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5581 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5641 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5701 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5761 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5821 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5881 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5941 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6001 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6061 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6121 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     6181 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     6241 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6301 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6361 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6421 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6481 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6541 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6601 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6661 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6721 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     6781 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6841 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6901 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6961 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     7021 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7081 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     7141 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     7201 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     7261 ctcacatgt
//
