LOCUS       sequence-81343-e        5690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses splitTVA-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5690)
  AUTHORS   Kohara K, Pignatelli M, Rivest AJ, Jung HY, Kitamura T, Suh J, Frank
            D, Kajikawa K, Mise N, Obata Y, Wickersham IR, Tonegawa S
  TITLE     Cell type-specific genetic and optogenetic tools reveal hippocampal 
            CA2 circuits.
  JOURNAL   Nat Neurosci. 2014 Feb;17(2):269-79. doi: 10.1038/nn.3614. Epub 2013
            Dec 15.
  PUBMED    24336151
REFERENCE   2  (bases 1 to 5690)
  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"; doi: "10.1038/nn.3614"; 
            journalName: "Nat Neurosci"; date: "2014-02"; volume: "17"; issue: 
            "2"; pages: "269-79"
COMMENT     Sequence Label: pAAV-synP-FLEX-splitTVA-EGFP
FEATURES             Location/Qualifiers
     source          1..5690
                     /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        158..605
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      612..621
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    627..660
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    711..744
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             complement(join(753..1466,1467..1469,1470..1472))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(756..1466,1467..1469,1470..1472))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(798..819)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1147..1166
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1406..1427
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1473..1529)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     protein_bind    complement(2012..2045)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2096..2129)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     misc_feature    2131..2719
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2184..2204)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2602..2613)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2727..2951
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2953..3093
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2953..3082
                     /label=AAV2 ITR
     rep_origin      3168..3623
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3255..3274)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3465..3486
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3640..3659)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3759..3781
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3819..3837)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3905..4009
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4010..4078,4079..4870)
                     /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(4228..4247)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5041..5629
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5530..5549
                     /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 gctagactgc agagggccct gcgtatgagt gcaagtgggt tttaggacca
      181 ggatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc
      241 caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc
      301 ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg
      361 gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc
      421 cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac
      481 cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg
      541 gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag
      601 cgcagtcagc cgccaccatg gttaatataa cttcgtataa tgtatgctat acgaagttat
      661 gcagaatggt agctggattg tagctgctat tagcaatatg aaacctctta ataacttcgt
      721 ataaagtatc ctatacgaag ttatggcgcg ccttacttgt acagctcgtc catgccgaga
      781 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg
      841 tctttgctca gggcggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg
      901 tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg
      961 ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag
     1021 acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg
     1081 aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg
     1141 gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct
     1201 tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag
     1261 cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg
     1321 gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg
     1381 gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc
     1441 ccggtgaaca gctcctcgcc cttgctcacc atgggtccag ggttctcctc cacgtctcca
     1501 gcctgcttca gcaggctgaa gttagtagct ccggatccgg agaacaagtc tgcctggctc
     1561 ttgtcaggca ccagcagctc cttggatgcg ctttcaagac tgaagatgtc agacctgctt
     1621 tttgctttgg acttccccca tgcagcgata ccacccacag ctaccaggca gcacaggagc
     1681 actgcagtga tcagcatcca catgcggccg tgattcctgg ctggcagagc acgtccagga
     1741 gcaggggcag tgggagcctc tgtgccgttg tccgtgggca ccgcggggat cgcgctggtc
     1801 ccgcagcccc actcgtcccg cccgtcgtcg cagtcggggt gtccgtcgca cagccagtcc
     1861 tgcgggtaac actccccgtg ggcaccgggc ggctccgagc agcggaactg accggggggg
     1921 caacgggaca aagaaccgtt accggacccg ttaccggtca cgttaccggg cagcagcagc
     1981 agcagcagcg cgggcagcag ccgcgcacct aataacttcg tatagcatac attatacgaa
     2041 gttatttgcc ttaacccaga aattatcact gttattcttt agaatggtgc aaagaataac
     2101 ttcgtatagg atactttata cgaagttatg aatcaacctc tggattacaa aatttgtgaa
     2161 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2221 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2281 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2341 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2401 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     2461 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     2521 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     2581 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     2641 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     2701 ctttgggccg cctccccgcc tggatcctgt gccttctagt tgccagccat ctgttgtttg
     2761 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2821 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2881 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     2941 gggctctatg gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     3001 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     3061 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     3121 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     3181 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3241 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3301 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3361 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     3421 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3481 aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc
     3541 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3601 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     3661 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3721 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3781 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3841 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3901 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3961 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4021 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4081 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4141 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4201 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4261 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4321 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4381 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4441 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4501 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4561 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4621 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4681 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4741 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4801 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4861 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4921 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4981 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5041 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5101 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5161 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     5221 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5281 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5341 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5401 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5461 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5521 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5581 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5641 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
