LOCUS       sequence-215866-        7497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Flpo recombinase-dependent GtACR2-Kv2.1C-linker-TlcnC, 
            which targets GtACR2 to the somatodendritic compartment. Messier et 
            al found this hybrid construct was the most effective at targeting.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7497)
  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 7497)
  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-Kv2.1C-linker-T...
FEATURES             Location/Qualifiers
     source          1..7497
                     /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(1848..2558,2559..2561))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1890..1911)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2239..2258
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2498..2519
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(3564..3597)
                     /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    3624..4212
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3677..3697)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4095..4106)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4244..4720
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4443..4462)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4760..4900
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4760..4889
                     /label=AAV2 ITR
     rep_origin      4975..5430
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5062..5081)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5272..5293
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5447..5466)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5566..5588
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5626..5644)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5712..5816
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5817..5885,5886..6677)
                     /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(6035..6054)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6848..7436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7337..7356
                     /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 ggcgcgcctc aggaagatgt cagctggata gcgaaaacct cgccatctgc aggtgactct
     1621 gcagagcctg atcctgaacc gctaccgctt ccaaaccttg tagcttcggg gaagtcggtg
     1681 gcgcaggaaa tgaagctatc gatactagac atgcttctca tgtcaatcac gccctcggtc
     1741 ctagctggca gaggagccac tggagatggc atggagctca tttccagttc ctcagggggt
     1801 ttactctgag gggccatctc cttagtgttc aggatgggct gagactgctt gtacagctcg
     1861 tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc
     1921 ttctcgttgg ggtctttgct cagggcggac tggtagctca ggtagtggtt gtcgggcagc
     1981 agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg
     2041 ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg
     2101 gccatgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg
     2161 ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg
     2221 aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc
     2281 tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg
     2341 tagcgggcga agcactgcag gccgtagccg aaggtggtca cgagggtggg ccagggcacg
     2401 ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc
     2461 tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg
     2521 atgggcacca ccccggtgaa cagctcctcg cccttgctca cggcggccgc cctgccgaac
     2581 attctgatgt tgacctcgtc gttggggggg gcattcttct cgtggttcag tccgaagggg
     2641 tcgtcggcct caggcttctc ctggccaggg aggcaagccg ggtcccactt tccctcgagc
     2701 actccccagg cggtggacca caggatcacg ccgtaggtgt tcttgcacag gccgtcggcg
     2761 agcagataca ggatggcgct cacgttctcg tggatcacgc acacgccggt gggggaaatc
     2821 agccacacga tggggaacat ggtccaggcg atgaagaaga tcagcctcag catcttgatc
     2881 ctcaccacca ccttctgggc cagaggggtg cccacggact cgaagccgta cagtcccacc
     2941 tggaagatgg tgaacaccac gctgtattcg aagaccaggc aggtagcgcc gaagaagaag
     3001 aacagccact tcatgtactc ggcgggggag atggtagcgg taatgcccat caccaccctg
     3061 ataatgctgg cggcctgggc gatggggttc agagggatgc tcttgtactt caccagagcc
     3121 atgttggaga tctggcccag cataatgggg catgtgcaca gccagctggc catcctggac
     3181 cagggaaaga acctgccgtc cctcatctgc agtgtgccat tgccggtgct ggccagggcg
     3241 tatgtgatgg actccacgaa gggcaggtac acgctctccc agggcacctt gggggccctg
     3301 ctgatcatga agatggtctg gcatccgctc accacggccc acaccagctg cagcaggctc
     3361 acgaaggtgc tgtcgatcct gctcatgttg tagcactctg tgtgggtgct ggcccactcg
     3421 ccgtacacga cctgggaggc catggatccc ggctagctac gtaggcgcgc cttaattaag
     3481 aagttcctat acttctgata gaataggaac ttcttgcctt aacccagaaa ttatcactgt
     3541 tattctttag aatggtgcaa agagaagttc ctatactttc tagagaatag gaacttccag
     3601 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     3661 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     3721 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     3781 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     3841 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     3901 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     3961 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     4021 catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct
     4081 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     4141 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     4201 ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga
     4261 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     4321 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     4381 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     4441 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     4501 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     4561 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     4621 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     4681 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac
     4741 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     4801 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     4861 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     4921 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     4981 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5041 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5101 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5161 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     5221 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5281 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     5341 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5401 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     5461 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5521 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5581 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     5641 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     5701 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5761 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5821 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5881 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5941 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6001 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6061 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6121 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6181 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6241 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6301 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6361 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6421 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6481 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6541 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6601 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6661 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6721 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6781 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6841 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6901 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6961 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     7021 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7081 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7141 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7201 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7261 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7321 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7381 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7441 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
