LOCUS       sequence-220121-        7457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Flpo recombinase-dependent GtACR2-Kv2.1C-TlcnC, which 
            targets GtACR2 to the somatodendritic compartment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7457)
  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 7457)
  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-TlcnC
FEATURES             Location/Qualifiers
     source          1..7457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   122..251
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        352..1530
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          583..1521
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1478..1498
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1621..1654
                     /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(1929..2639,2640..2642))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1971..1992)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2320..2339
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2579..2600
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(3645..3678)
                     /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    3705..4293
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3758..3778)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4176..4187)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4325..4801
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4524..4543)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4841..4981
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4841..4970
                     /label=AAV2 ITR
     rep_origin      5056..5511
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5143..5162)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5353..5374
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5528..5547)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5647..5669
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5707..5725)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5793..5897
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5898..5966,5967..6758)
                     /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(6116..6135)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6929..7457,1..60)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7418..7437
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
       61 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      121 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
      181 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      241 taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt ttaaacagag
      301 aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt
      361 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
      421 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
      481 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
      541 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt
      601 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc
      661 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg
      721 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc
      781 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag
      841 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt
      901 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc
      961 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca
     1021 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg
     1081 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga
     1141 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc
     1201 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc
     1261 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg
     1321 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc
     1381 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg
     1441 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa
     1501 agtttttttc ttccatttca ggtgtcgtga ggtaccgaag ttcctattct atcagaagta
     1561 taggaacttc gcagaatggt agctggattg tagctgctat tagcaatatg aaacctctta
     1621 gaagttccta ttctctagaa agtataggaa cttcgaattc ttacttgtac aggcgcgcct
     1681 caggaagatg tcagctggat agcgaaaacc tcgccatctg caggtgactc tgcaaacctt
     1741 gtagcttcgg ggaagtcggt ggcgcaggaa atgaagctat cgatactaga catgcttctc
     1801 atgtcaatca cgccctcggt cctagctggc agaggagcca ctggagatgg catggagctc
     1861 atttccagtt cctcaggggg tttactctga ggggccatct ccttagtgtt caggatgggc
     1921 tgagactgct tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
     1981 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctggtagctc
     2041 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
     2101 tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg
     2161 atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc
     2221 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg
     2281 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc
     2341 ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg
     2401 tgctgcttca tgtggtcggg gtagcgggcg aagcactgca ggccgtagcc gaaggtggtc
     2461 acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc
     2521 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg
     2581 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc
     2641 acggcggccg ccctgccgaa cattctgatg ttgacctcgt cgttgggggg ggcattcttc
     2701 tcgtggttca gtccgaaggg gtcgtcggcc tcaggcttct cctggccagg gaggcaagcc
     2761 gggtcccact ttccctcgag cactccccag gcggtggacc acaggatcac gccgtaggtg
     2821 ttcttgcaca ggccgtcggc gagcagatac aggatggcgc tcacgttctc gtggatcacg
     2881 cacacgccgg tgggggaaat cagccacacg atggggaaca tggtccaggc gatgaagaag
     2941 atcagcctca gcatcttgat cctcaccacc accttctggg ccagaggggt gcccacggac
     3001 tcgaagccgt acagtcccac ctggaagatg gtgaacacca cgctgtattc gaagaccagg
     3061 caggtagcgc cgaagaagaa gaacagccac ttcatgtact cggcggggga gatggtagcg
     3121 gtaatgccca tcaccaccct gataatgctg gcggcctggg cgatggggtt cagagggatg
     3181 ctcttgtact tcaccagagc catgttggag atctggccca gcataatggg gcatgtgcac
     3241 agccagctgg ccatcctgga ccagggaaag aacctgccgt ccctcatctg cagtgtgcca
     3301 ttgccggtgc tggccagggc gtatgtgatg gactccacga agggcaggta cacgctctcc
     3361 cagggcacct tgggggccct gctgatcatg aagatggtct ggcatccgct caccacggcc
     3421 cacaccagct gcagcaggct cacgaaggtg ctgtcgatcc tgctcatgtt gtagcactct
     3481 gtgtgggtgc tggcccactc gccgtacacg acctgggagg ccatggatcc cggctagcta
     3541 cgtaggcgcg ccttaattaa gaagttccta tacttctgat agaataggaa cttcttgcct
     3601 taacccagaa attatcactg ttattcttta gaatggtgca aagagaagtt cctatacttt
     3661 ctagagaata ggaacttcca gaattcgata tcaagcttat cgataatcaa cctctggatt
     3721 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3781 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3841 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3901 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3961 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     4021 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     4081 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
     4141 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     4201 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     4261 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat
     4321 ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     4381 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     4441 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     4501 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     4561 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     4621 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     4681 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     4741 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     4801 tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     4861 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     4921 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     4981 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     5041 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5101 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     5161 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5221 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     5281 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5341 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     5401 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     5461 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     5521 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     5581 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     5641 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     5701 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     5761 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5821 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5881 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5941 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     6001 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     6061 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     6121 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     6181 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6241 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6301 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6361 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6421 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6481 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6541 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6601 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6661 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6721 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6781 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6841 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6901 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6961 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     7021 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     7081 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7141 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7201 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7261 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7321 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7381 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7441 gtcctgtcgg gtttcgc
//
