LOCUS       sequence-215863-        7272 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Flpo recombinase-dependent GtACR2-TlcnC, which targets 
            GtACR2 to the somatodendritic compartment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7272)
  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 7272)
  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-TlcnC
FEATURES             Location/Qualifiers
     source          1..7272
                     /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(1623..2333,2334..2336))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1665..1686)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2014..2033
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2273..2294
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(3339..3372)
                     /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    3399..3987
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3452..3472)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3870..3881)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4019..4495
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4218..4237)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4535..4675
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4535..4664
                     /label=AAV2 ITR
     rep_origin      4750..5205
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4837..4856)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5047..5068
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5222..5241)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5341..5363
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5401..5419)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5487..5591
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5592..5660,5661..6452)
                     /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(5810..5829)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6623..7211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7112..7131
                     /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 aggaagatgt cagctggata gcgaaaacct cgccatctgc aggtgactct
     1621 gccttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     1681 accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactggta gctcaggtag
     1741 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg
     1801 gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg
     1861 ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg
     1921 tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc
     1981 agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag
     2041 aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc
     2101 ttcatgtggt cggggtagcg ggcgaagcac tgcaggccgt agccgaaggt ggtcacgagg
     2161 gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg
     2221 taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg
     2281 tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcacggcg
     2341 gccgccctgc cgaacattct gatgttgacc tcgtcgttgg ggggggcatt cttctcgtgg
     2401 ttcagtccga aggggtcgtc ggcctcaggc ttctcctggc cagggaggca agccgggtcc
     2461 cactttccct cgagcactcc ccaggcggtg gaccacagga tcacgccgta ggtgttcttg
     2521 cacaggccgt cggcgagcag atacaggatg gcgctcacgt tctcgtggat cacgcacacg
     2581 ccggtggggg aaatcagcca cacgatgggg aacatggtcc aggcgatgaa gaagatcagc
     2641 ctcagcatct tgatcctcac caccaccttc tgggccagag gggtgcccac ggactcgaag
     2701 ccgtacagtc ccacctggaa gatggtgaac accacgctgt attcgaagac caggcaggta
     2761 gcgccgaaga agaagaacag ccacttcatg tactcggcgg gggagatggt agcggtaatg
     2821 cccatcacca ccctgataat gctggcggcc tgggcgatgg ggttcagagg gatgctcttg
     2881 tacttcacca gagccatgtt ggagatctgg cccagcataa tggggcatgt gcacagccag
     2941 ctggccatcc tggaccaggg aaagaacctg ccgtccctca tctgcagtgt gccattgccg
     3001 gtgctggcca gggcgtatgt gatggactcc acgaagggca ggtacacgct ctcccagggc
     3061 accttggggg ccctgctgat catgaagatg gtctggcatc cgctcaccac ggcccacacc
     3121 agctgcagca ggctcacgaa ggtgctgtcg atcctgctca tgttgtagca ctctgtgtgg
     3181 gtgctggccc actcgccgta cacgacctgg gaggccatgg atcccggcta gctacgtagg
     3241 cgcgccttaa ttaagaagtt cctatacttc tgatagaata ggaacttctt gccttaaccc
     3301 agaaattatc actgttattc tttagaatgg tgcaaagaga agttcctata ctttctagag
     3361 aataggaact tccagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa
     3421 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     3481 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     3541 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     3601 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     3661 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     3721 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     3781 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc
     3841 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     3901 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3961 ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg
     4021 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     4081 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     4141 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     4201 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     4261 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     4321 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     4381 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     4441 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     4501 tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     4561 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     4621 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     4681 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     4741 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4801 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4861 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4921 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     4981 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5041 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt
     5101 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5161 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     5221 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     5281 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     5341 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     5401 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     5461 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5521 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5581 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5641 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5701 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5761 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5821 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5881 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5941 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6001 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6061 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6121 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6181 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6241 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6301 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6361 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6421 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6481 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6541 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6601 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6661 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6721 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     6781 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6841 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6901 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6961 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7021 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7081 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7141 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7201 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7261 ttgctcacat gt
//
