LOCUS       sequence-220119-        7402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Flpo recombinase-dependent GtACR2-Kv2.1C, which targets 
            GtACR2 to the somatodendritic compartment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7402)
  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 7402)
  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
FEATURES             Location/Qualifiers
     source          1..7402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   92..221
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        322..1500
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          553..1491
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1448..1468
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1591..1624
                     /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(1844..2554,2555..2557))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1886..1907)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2235..2254
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2494..2515
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(3560..3593)
                     /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    3620..4208
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3673..3693)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4091..4102)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4240..4716
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4439..4458)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4756..4896
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4756..4885
                     /label=AAV2 ITR
     rep_origin      4971..5426
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5058..5077)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5268..5289
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5443..5462)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5562..5584
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5622..5640)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5708..5812
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5813..5881,5882..6673)
                     /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(6031..6050)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6844..7402,1..30)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7333..7352
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
       61 ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc
      121 actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
      181 gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca cgcgtaagct
      241 ttgcaaagat ggataaagtt ttaaacagag aggaatcttt gcagctaatg gaccttctag
      301 gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg cacatcgccc
      361 acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg
      421 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
      481 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
      541 gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat
      601 ggcccttgcg tgccttgaat tacttccacc tggctgcagt acgtgattct tgatcccgag
      661 cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct
      721 cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac
      781 cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct
      841 gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact
      901 ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt
      961 tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct
     1021 ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca
     1081 aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct
     1141 gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca
     1201 caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg
     1261 gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg
     1321 ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg
     1381 ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg
     1441 ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtga
     1501 ggtaccgaag ttcctattct atcagaagta taggaacttc gcagaatggt agctggattg
     1561 tagctgctat tagcaatatg aaacctctta gaagttccta ttctctagaa agtataggaa
     1621 cttcgaattc ttacttgtac gcgccttaaa accttgtagc ttcggggaag tcggtggcgc
     1681 aggaaatgaa gctatcgata ctagacatgc ttctcatgtc aatcacgccc tcggtcctag
     1741 ctggcagagg agccactgga gatggcatgg agctcatttc cagttcctca gggggtttac
     1801 tctgaggggc catctcctta gtgttcagga tgggctgaga ctgcttgtac agctcgtcca
     1861 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     1921 cgttggggtc tttgctcagg gcggactggt agctcaggta gtggttgtcg ggcagcagca
     1981 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt
     2041 cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca
     2101 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     2161 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     2221 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     2281 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     2341 gggcgaagca ctgcaggccg tagccgaagg tggtcacgag ggtgggccag ggcacgggca
     2401 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     2461 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     2521 gcaccacccc ggtgaacagc tcctcgccct tgctcacggc ggccgccctg ccgaacattc
     2581 tgatgttgac ctcgtcgttg gggggggcat tcttctcgtg gttcagtccg aaggggtcgt
     2641 cggcctcagg cttctcctgg ccagggaggc aagccgggtc ccactttccc tcgagcactc
     2701 cccaggcggt ggaccacagg atcacgccgt aggtgttctt gcacaggccg tcggcgagca
     2761 gatacaggat ggcgctcacg ttctcgtgga tcacgcacac gccggtgggg gaaatcagcc
     2821 acacgatggg gaacatggtc caggcgatga agaagatcag cctcagcatc ttgatcctca
     2881 ccaccacctt ctgggccaga ggggtgccca cggactcgaa gccgtacagt cccacctgga
     2941 agatggtgaa caccacgctg tattcgaaga ccaggcaggt agcgccgaag aagaagaaca
     3001 gccacttcat gtactcggcg ggggagatgg tagcggtaat gcccatcacc accctgataa
     3061 tgctggcggc ctgggcgatg gggttcagag ggatgctctt gtacttcacc agagccatgt
     3121 tggagatctg gcccagcata atggggcatg tgcacagcca gctggccatc ctggaccagg
     3181 gaaagaacct gccgtccctc atctgcagtg tgccattgcc ggtgctggcc agggcgtatg
     3241 tgatggactc cacgaagggc aggtacacgc tctcccaggg caccttgggg gccctgctga
     3301 tcatgaagat ggtctggcat ccgctcacca cggcccacac cagctgcagc aggctcacga
     3361 aggtgctgtc gatcctgctc atgttgtagc actctgtgtg ggtgctggcc cactcgccgt
     3421 acacgacctg ggaggccatg gatcccggct agctacgtag gcgcgcctta attaagaagt
     3481 tcctatactt ctgatagaat aggaacttct tgccttaacc cagaaattat cactgttatt
     3541 ctttagaatg gtgcaaagag aagttcctat actttctaga gaataggaac ttccagaatt
     3601 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3661 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3721 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3781 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3841 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3901 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3961 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     4021 gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg
     4081 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     4141 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     4201 ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc
     4261 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     4321 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     4381 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     4441 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     4501 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
     4561 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     4621 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
     4681 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgc
     4741 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     4801 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     4861 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     4921 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     4981 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5041 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5101 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5161 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     5221 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5281 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     5341 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     5401 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     5461 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     5521 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5581 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     5641 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     5701 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5761 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5821 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5881 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5941 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6001 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     6061 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6121 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     6181 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     6241 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6301 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     6361 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6421 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6481 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6541 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6601 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6661 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6721 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6781 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6841 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6901 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6961 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     7021 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7081 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7141 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7201 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     7261 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7321 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7381 acttgagcgt cgatttttgt ga
//
