LOCUS       sequence-215864-        7413 bp    DNA     circular SYN 15-APR-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 7413)
  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 7413)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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..7413
                     /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(1764..2474,2475..2477))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1806..1827)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2155..2174
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2414..2435
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(3480..3513)
                     /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    3540..4128
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3593..3613)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4011..4022)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4160..4636
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4359..4378)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4676..4816
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4676..4805
                     /label=AAV2 ITR
     rep_origin      4891..5346
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4978..4997)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5188..5209
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5363..5382)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5482..5504
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5542..5560)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5628..5732
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5733..5801,5802..6593)
                     /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(5951..5970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6764..7352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7253..7272
                     /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 tacttgtacc
     1561 gcgccttaaa accttgtagc ttcggggaag tcggtggcgc aggaaatgaa gctatcgata
     1621 ctagacatgc ttctcatgtc aatcacgccc tcggtcctag ctggcagagg agccactgga
     1681 gatggcatgg agctcatttc cagttcctca gggggtttac tctgaggggc catctcctta
     1741 gtgttcagga tgggctgaga ctgcttgtac agctcgtcca tgccgagagt gatcccggcg
     1801 gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg
     1861 gcggactggt agctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg
     1921 gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc
     1981 ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg
     2041 ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc
     2101 ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg
     2161 tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg
     2221 gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc gggcgaagca ctgcaggccg
     2281 tagccgaagg tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg
     2341 aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac
     2401 ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc
     2461 tcctcgccct tgctcacggc ggccgccctg ccgaacattc tgatgttgac ctcgtcgttg
     2521 gggggggcat tcttctcgtg gttcagtccg aaggggtcgt cggcctcagg cttctcctgg
     2581 ccagggaggc aagccgggtc ccactttccc tcgagcactc cccaggcggt ggaccacagg
     2641 atcacgccgt aggtgttctt gcacaggccg tcggcgagca gatacaggat ggcgctcacg
     2701 ttctcgtgga tcacgcacac gccggtgggg gaaatcagcc acacgatggg gaacatggtc
     2761 caggcgatga agaagatcag cctcagcatc ttgatcctca ccaccacctt ctgggccaga
     2821 ggggtgccca cggactcgaa gccgtacagt cccacctgga agatggtgaa caccacgctg
     2881 tattcgaaga ccaggcaggt agcgccgaag aagaagaaca gccacttcat gtactcggcg
     2941 ggggagatgg tagcggtaat gcccatcacc accctgataa tgctggcggc ctgggcgatg
     3001 gggttcagag ggatgctctt gtacttcacc agagccatgt tggagatctg gcccagcata
     3061 atggggcatg tgcacagcca gctggccatc ctggaccagg gaaagaacct gccgtccctc
     3121 atctgcagtg tgccattgcc ggtgctggcc agggcgtatg tgatggactc cacgaagggc
     3181 aggtacacgc tctcccaggg caccttgggg gccctgctga tcatgaagat ggtctggcat
     3241 ccgctcacca cggcccacac cagctgcagc aggctcacga aggtgctgtc gatcctgctc
     3301 atgttgtagc actctgtgtg ggtgctggcc cactcgccgt acacgacctg ggaggccatg
     3361 gatcccggct agctacgtag gcgcgcctta attaagaagt tcctatactt ctgatagaat
     3421 aggaacttct tgccttaacc cagaaattat cactgttatt ctttagaatg gtgcaaagag
     3481 aagttcctat actttctaga gaataggaac ttccagaatt cgatatcaag cttatcgata
     3541 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3601 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3661 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3721 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3781 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3841 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3901 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3961 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     4021 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     4081 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag
     4141 cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     4201 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     4261 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     4321 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     4381 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     4441 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     4501 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     4561 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     4621 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa
     4681 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     4741 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     4801 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4861 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     4921 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4981 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5041 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5101 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5161 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5221 accctatctc gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     5281 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     5341 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     5401 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     5461 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     5521 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     5581 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5641 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5701 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5761 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5821 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5881 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5941 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6001 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6061 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6121 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6181 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6241 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6301 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6361 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6421 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6481 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6541 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6601 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6661 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6721 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6781 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6841 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6901 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6961 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7021 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7081 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7141 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7201 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7261 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7321 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7381 tcctggcctt ttgctggcct tttgctcaca tgt
//
