LOCUS       sequence-220133-        7262 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 7262)
  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 7262)
  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..7262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1500..1533
                     /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(1613..2323,2324..2326))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1655..1676)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2004..2023
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2263..2284
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(3329..3362)
                     /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    3389..3977
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3442..3462)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3860..3871)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4009..4485
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4208..4227)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4525..4665
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4525..4654
                     /label=AAV2 ITR
     rep_origin      4740..5195
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4827..4846)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5037..5058
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5212..5231)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5331..5353
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5391..5409)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5477..5581
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5582..5650,5651..6442)
                     /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(5800..5819)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6613..7201
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7102..7121
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccgaagt tcctattcta tcagaagtat
     1441 aggaacttcg cagaatggta gctggattgt agctgctatt agcaatatga aacctcttag
     1501 aagttcctat tctctagaaa gtataggaac ttcgaattct tacttgtaca gtttaaactc
     1561 aggaagatgt cagctggata gcgaaaacct cgccatctgc aggtgactct gccttgtaca
     1621 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
     1681 cgcgcttctc gttggggtct ttgctcaggg cggactggta gctcaggtag tggttgtcgg
     1741 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
     1801 cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct
     1861 tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga
     1921 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
     1981 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc
     2041 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
     2101 cggggtagcg ggcgaagcac tgcaggccgt agccgaaggt ggtcacgagg gtgggccagg
     2161 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat
     2221 cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     2281 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcacggcg gccgccctgc
     2341 cgaacattct gatgttgacc tcgtcgttgg ggggggcatt cttctcgtgg ttcagtccga
     2401 aggggtcgtc ggcctcaggc ttctcctggc cagggaggca agccgggtcc cactttccct
     2461 cgagcactcc ccaggcggtg gaccacagga tcacgccgta ggtgttcttg cacaggccgt
     2521 cggcgagcag atacaggatg gcgctcacgt tctcgtggat cacgcacacg ccggtggggg
     2581 aaatcagcca cacgatgggg aacatggtcc aggcgatgaa gaagatcagc ctcagcatct
     2641 tgatcctcac caccaccttc tgggccagag gggtgcccac ggactcgaag ccgtacagtc
     2701 ccacctggaa gatggtgaac accacgctgt attcgaagac caggcaggta gcgccgaaga
     2761 agaagaacag ccacttcatg tactcggcgg gggagatggt agcggtaatg cccatcacca
     2821 ccctgataat gctggcggcc tgggcgatgg ggttcagagg gatgctcttg tacttcacca
     2881 gagccatgtt ggagatctgg cccagcataa tggggcatgt gcacagccag ctggccatcc
     2941 tggaccaggg aaagaacctg ccgtccctca tctgcagtgt gccattgccg gtgctggcca
     3001 gggcgtatgt gatggactcc acgaagggca ggtacacgct ctcccagggc accttggggg
     3061 ccctgctgat catgaagatg gtctggcatc cgctcaccac ggcccacacc agctgcagca
     3121 ggctcacgaa ggtgctgtcg atcctgctca tgttgtagca ctctgtgtgg gtgctggccc
     3181 actcgccgta cacgacctgg gaggccatgg atcccggcta gctacgtagg cgcgccttaa
     3241 ttaagaagtt cctatacttc tgatagaata ggaacttctt gccttaaccc agaaattatc
     3301 actgttattc tttagaatgg tgcaaagaga agttcctata ctttctagag aataggaact
     3361 tccagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     3421 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3481 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3541 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3601 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3661 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3721 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3781 gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac
     3841 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     3901 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     3961 ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc
     4021 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     4081 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     4141 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     4201 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     4261 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     4321 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     4381 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     4441 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta
     4501 accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     4561 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     4621 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     4681 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     4741 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4801 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4861 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4921 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     4981 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5041 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     5101 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5161 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     5221 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     5281 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     5341 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     5401 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     5461 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5521 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5581 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5641 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5701 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5761 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5821 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5881 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5941 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6001 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6061 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6121 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6181 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6241 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6301 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6361 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6421 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6481 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6541 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6601 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6661 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6721 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6781 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6841 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6901 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6961 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7021 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7081 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7141 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7201 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7261 gt
//
