LOCUS       Exported                7307 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  expresses Flpo recombinase-dependent GtACR2-Nlgn1C, which targets 
            GtACR2 to the somatodendritic compartment.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-EF1-alpha-FRT-FLEX-GtACR2-EYFP-Nlgn1C
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7307)
  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 7307)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7307
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   48..177
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        278..1456
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          509..1447
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1404..1424
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1547..1580
                     /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(1705..2418)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1747..1768)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2096..2115
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2355..2376
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(3421..3454)
                     /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    3481..4069
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3534..3554)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3952..3963)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4101..4577
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4300..4319)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4617..4757
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4617..4746
                     /label=AAV2 ITR
     rep_origin      4832..5287
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4919..4938)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5129..5150
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5304..5323)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5423..5445
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5483..5501)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5569..5673
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5674..6534
                     /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(5892..5911)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6705..7293
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7194..7213
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct
       61 gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct
      121 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg
      181 gccgcacgcg taagctttgc aaagatggat aaagttttaa acagagagga atctttgcag
      241 ctaatggacc ttctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc
      301 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg
      361 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
      421 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
      481 tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg
      541 cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc tgcagtacgt
      601 gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc cttgcgctta
      661 aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg gccgccgcgt
      721 gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc tagccattta
      781 aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg taaatgcggg
      841 ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc
      901 gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga gaatcggacg
      961 ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc
     1021 cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc
     1081 gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg gagagcgggc
     1141 gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga
     1201 ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct tttggagtac
     1261 gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt
     1321 ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt
     1381 tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc
     1441 atttcaggtg tcgtgaggta ccgaagttcc tattctatca gaagtatagg aacttcgcag
     1501 aatggtagct ggattgtagc tgctattagc aatatgaaac ctcttagaag ttcctattct
     1561 ctagaaagta taggaacttc gaattcttac ttgtacagtt taaactcaca tggttatggt
     1621 attgggtgtc atcaggggaa tgtcgtctgg gcttcggcgc attgccaggg tatagtccgg
     1681 agggcatgct gtcctcagca ccaccttgta cagctcgtcc atgccgagag tgatcccggc
     1741 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
     1801 ggcggactgg tagctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
     1861 ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt tgtggcggat
     1921 cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct
     1981 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
     2041 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
     2101 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
     2161 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cgggcgaagc actgcaggcc
     2221 gtagccgaag gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
     2281 gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
     2341 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
     2401 ctcctcgccc ttgctcacgg cggccgccct gccgaacatt ctgatgttga cctcgtcgtt
     2461 ggggggggca ttcttctcgt ggttcagtcc gaaggggtcg tcggcctcag gcttctcctg
     2521 gccagggagg caagccgggt cccactttcc ctcgagcact ccccaggcgg tggaccacag
     2581 gatcacgccg taggtgttct tgcacaggcc gtcggcgagc agatacagga tggcgctcac
     2641 gttctcgtgg atcacgcaca cgccggtggg ggaaatcagc cacacgatgg ggaacatggt
     2701 ccaggcgatg aagaagatca gcctcagcat cttgatcctc accaccacct tctgggccag
     2761 aggggtgccc acggactcga agccgtacag tcccacctgg aagatggtga acaccacgct
     2821 gtattcgaag accaggcagg tagcgccgaa gaagaagaac agccacttca tgtactcggc
     2881 gggggagatg gtagcggtaa tgcccatcac caccctgata atgctggcgg cctgggcgat
     2941 ggggttcaga gggatgctct tgtacttcac cagagccatg ttggagatct ggcccagcat
     3001 aatggggcat gtgcacagcc agctggccat cctggaccag ggaaagaacc tgccgtccct
     3061 catctgcagt gtgccattgc cggtgctggc cagggcgtat gtgatggact ccacgaaggg
     3121 caggtacacg ctctcccagg gcaccttggg ggccctgctg atcatgaaga tggtctggca
     3181 tccgctcacc acggcccaca ccagctgcag caggctcacg aaggtgctgt cgatcctgct
     3241 catgttgtag cactctgtgt gggtgctggc ccactcgccg tacacgacct gggaggccat
     3301 ggatcccggc tagctacgta ggcgcgcctt aattaagaag ttcctatact tctgatagaa
     3361 taggaacttc ttgccttaac ccagaaatta tcactgttat tctttagaat ggtgcaaaga
     3421 gaagttccta tactttctag agaataggaa cttccagaat tcgatatcaa gcttatcgat
     3481 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3541 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3601 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3661 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     3721 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3781 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3841 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     3901 gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3961 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4021 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga
     4081 gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     4141 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     4201 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     4261 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga
     4321 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc
     4381 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt
     4441 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat
     4501 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg
     4561 ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga
     4621 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4681 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     4741 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4801 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     4861 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     4921 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4981 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5041 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5101 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5161 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     5221 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     5281 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     5341 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     5401 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     5461 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5521 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5581 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5641 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5701 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5761 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5821 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5881 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5941 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     6001 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     6061 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     6121 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6181 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     6241 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6301 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6361 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6421 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6481 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6541 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6601 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6661 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6721 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6781 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6841 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6901 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6961 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7021 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7081 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7141 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7201 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7261 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgc
//
