LOCUS       sequence-217315-        7188 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid that expresses GFP-Fxr1 fusion protein under hSYN 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7188)
  AUTHORS   Khlghatyan J, Evstratova A, Chamberland S, Marakhovskaia A, 
            Bahremand A, Toth K, Beaulieu JM
  TITLE     Mental Illnesses-Associated Fxr1 and Its Negative Regulator Gsk3beta
            Are Modulators of Anxiety and Glutamatergic Neurotransmission.
  JOURNAL   Front Mol Neurosci. 2018 Apr 12;11:119. doi: 
            10.3389/fnmol.2018.00119. eCollection 2018.
  PUBMED    29706865
REFERENCE   2  (bases 1 to 7188)
  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: "Front Mol 
            Neurosci."; date: "2018-04-12"; volume: "11:119. doi"; pages: " 
            10.3389/fnmol.2018.00119. eCollection 2018"
COMMENT     Sequence Label: pAAV hSyn GFP-Fxr1
FEATURES             Location/Qualifiers
     source          1..7188
                     /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        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      645..654
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(651..653,654..656,657..1367)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(696..717)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(957..976)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1304..1325
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3315..3903
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3368..3388)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3786..3797)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3935..4411
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4134..4153)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4451..4591
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4451..4580
                     /label=AAV2 ITR
     rep_origin      4666..5121
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4753..4772)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4963..4984
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5138..5157)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5257..5279
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5317..5335)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5403..5507
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5508..5576,5577..6368)
                     /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(5726..5745)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6539..7127
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7028..7047
                     /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc cgccgccacc atggtgagca
      661 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      721 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      781 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      841 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
      901 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
      961 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1021 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1081 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     1141 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     1201 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1261 cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1321 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagagt gctggtggta
     1381 gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggtgcg gagctgacgg
     1441 tggaggttcg cggctccaac ggggctttct acaagggatt tatcaaagat gtccacgaag
     1501 actccctcac agttgttttt gaaaataatt ggcaaccaga acgccaggtt ccgtttaatg
     1561 aagtgcgatt accaccacca cctgatataa aaaaagaaat tagtgaagga gatgaagtag
     1621 aggtatattc aagagcaaat gaccaagagc catgtggatg gtggctggct aaagttcgga
     1681 tgatgaaagg cgagttttat gtcattgaat atgctgcttg tgatgccact tacaatgaaa
     1741 tagtcacatt tgaacgactt cggcctgtca atcaaaataa aactgtcaaa aaaaatacct
     1801 tctttaagtg cacagtggat gttcctgagg acctgagaga agcgtgtgct aatgaaaatg
     1861 cccataaaga ttttaagaaa gcagtaggag catgcagaat cttttatcat cctgaaacta
     1921 cccagctaat gatactgtcg gccagtgaag caactgtgaa gagagtaaat atcttaagtg
     1981 atatgcattt gagaagtatt cggacgaagt tgatgcttat gtccagaaat gaagaggcca
     2041 ctaagcattt agaatgcaca aaacaacttg cagcagcttt tcatgaagaa tttgttgtga
     2101 gagaagattt aatgggcctg gcgataggaa cgcatggcag taacatacag caagctagga
     2161 aggttcctgg agttactgcc attgagttag atgaagacac cggaacgttt agaatctatg
     2221 gagagagtgc tgaggctgta aaaaaagcta gaggtttctt ggaatttgtg gaagatttta
     2281 ttcaagttcc caggaatctt gttggaaaag taattggaaa aaatggcaaa gttattcaag
     2341 aaatagtgga taaatctggt gtggttcgag taagaattga aggagacaat gaaaataaac
     2401 tacctagaga agacggaatg gttccatttg tatttgttgg cactaaagaa agcattggga
     2461 atgtgcaagt tcttctagag tatcacatcg cttacttaaa ggaagtggaa caactaagaa
     2521 tggaacgtct gcagattgat gagcagctgc gacagattgg ttctaggtct tatagtggaa
     2581 gaggcagagg tcgtcggggc cctaattaca cctccggtta tggtacaaat tctgagctgt
     2641 ctaacccctc cgaaacagaa tctgagcgta aagatgagct gagtgattgg tcattggcag
     2701 gagaagatga tcgagagact cgacatcagc gagacagcag gagacgccca ggaggaagag
     2761 gcagaagtgt ttctggggga cgaggtcgtg gtggaccacg tggtggcaaa tcctccatca
     2821 gttctgtgct caaagatcca gacagcaatc catacagctt acttgataat acagaatccg
     2881 atcagactgc agacactgac gccagcgaat ctcaccacag tactaaccgt cgtaggcggt
     2941 ctcgtagacg gaggactgat gaagatgctg ttctgatgga tggactgact gaatctgata
     3001 cagcctcagt taatgagaat gggctagtca cagttgcaga ttatatttct agagctgagt
     3061 ctcagagcag acaaagaaac ctcccaaggg aaactttggc taaaaacaag aaagaaatgg
     3121 caaaagatgt gattgaagag catggtcctt cagaaaaggc aataaatggt ccaactagcg
     3181 cttctggcga tgaaattcct aagctaccgc gtactctggg agaagaaaag actaagacct
     3241 taaaagaaga cagcactcaa gaagcagcag tcctgaatgg tgtttcataa gaattcgata
     3301 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     3361 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     3421 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     3481 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     3541 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     3601 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     3661 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     3721 ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     3781 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     3841 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     3901 cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc
     3961 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     4021 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     4081 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     4141 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     4201 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     4261 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     4321 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     4381 ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc
     4441 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     4501 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     4561 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     4621 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     4681 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4741 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4801 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4861 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     4921 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4981 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     5041 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     5101 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     5161 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     5221 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     5281 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     5341 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     5401 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5461 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5521 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5581 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5641 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5701 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5761 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5821 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5881 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5941 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6001 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     6061 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     6121 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6181 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6241 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6301 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6361 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6421 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6481 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6541 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6601 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6661 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6721 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6781 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6841 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6901 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6961 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     7021 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     7081 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     7141 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
