LOCUS       sequence-79077-e        4960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Can be used to generate AAV virus that will express the FlpO 
            recombinase gene in neurons from the synapsin promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4960)
  AUTHORS   Madisen L, Garner AR, Shimaoka D, Chuong AS, Klapoetke NC, Li L, van
            der Bourg A, Niino Y, Egolf L, Monetti C, Gu H, Mills M, Cheng A, 
            Tasic B, Nguyen TN, Sunkin SM, Benucci A, Nagy A, Miyawaki A, 
            Helmchen F, Empson RM, Knopfel T, Boyden ES, Reid RC, Carandini M, 
            Zeng H
  TITLE     Transgenic mice for intersectional targeting of neural sensors and 
            effectors with high specificity and performance.
  JOURNAL   Neuron. 2015 Mar 4;85(5):942-58. doi: 10.1016/j.neuron.2015.02.022.
  PUBMED    25741722
REFERENCE   2  (bases 1 to 4960)
  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: 
            "10.1016/j.neuron.2015.02"; journalName: "Neuron"; date: "2015-03-4-
            4"; volume: "85"; issue: "5"; pages: "942-58"
COMMENT     Sequence Label: AAV phSyn1(S)-FlpO-bGHpA
FEATURES             Location/Qualifiers
     source          1..4960
                     /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        177..624
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      654..663
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(660..665,666..686,687..1958)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     primer_bind     complement(1987..2004)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1993..2200
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2223..2363
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2223..2352
                     /label=AAV2 ITR
     rep_origin      2438..2893
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2525..2544)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2735..2756
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2910..2929)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3029..3051
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3089..3107)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3175..3279
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3280..3348,3349..4140)
                     /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(3498..3517)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4311..4899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4800..4819
                     /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 cgcgtctgca gagggccctg cgtatgagtg
      181 caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg cctacctgac gaccgacccc
      241 gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg
      301 gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca
      361 gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg
      421 acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg cgtccgcgcc
      481 gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca cgggcgcgac
      541 catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg cagcggagga
      601 gtcgtgtcgt gcctgagagc gcagacgcgt ggacggtacc aagcttacgc gtagccacca
      661 tggctcctaa gaagaagagg aaggtgatga gccagttcga catcctgtgc aagacccccc
      721 ccaaggtgct ggtgcggcag ttcgtggaga gattcgagag gcccagcggc gagaagatcg
      781 ccagctgtgc cgccgagctg acctacctgt gctggatgat cacccacaac ggcaccgcca
      841 tcaagagggc caccttcatg agctacaaca ccatcatcag caacagcctg agcttcgaca
      901 tcgtgaacaa gagcctgcag ttcaagtaca agacccagaa ggccaccatc ctggaggcca
      961 gcctgaagaa gctgatcccc gcctgggagt tcaccatcat cccttacaac ggccagaagc
     1021 accagagcga catcaccgac atcgtgtcca gcctgcagct gcagttcgag agcagcgagg
     1081 aggccgacaa gggcaacagc cacagcaaga agatgctgaa ggccctgctg tccgagggcg
     1141 agagcatctg ggagatcacc gagaagatcc tgaacagctt cgagtacacc agcaggttca
     1201 ccaagaccaa gaccctgtac cagttcctgt tcctggccac attcatcaac tgcggcaggt
     1261 tcagcgacat caagaacgtg gaccccaaga gcttcaagct ggtgcagaac aagtacctgg
     1321 gcgtgatcat tcagtgcctg gtgaccgaga ccaagacaag cgtgtccagg cacatctact
     1381 ttttcagcgc cagaggcagg atcgaccccc tggtgtacct ggacgagttc ctgaggaaca
     1441 gcgagcccgt gctgaagaga gtgaacagga ccggcaacag cagcagcaac aagcaggagt
     1501 accagctgct gaaggacaac ctggtgcgca gctacaacaa ggccctgaag aagaacgccc
     1561 cctaccccat cttcgctatc aagaacggcc ctaagagcca catcggcagg cacctgatga
     1621 ccagctttct gagcatgaag ggcctgaccg agctgacaaa cgtggtgggc aactggagcg
     1681 acaagagggc ctccgccgtg gccaggacca cctacaccca ccagatcacc gccatccccg
     1741 accactactt cgccctggtg tccaggtact acgcctacga ccccatcagc aaggagatga
     1801 tcgccctgaa ggacgagacc aaccccatcg aggagtggca gcacatcgag cagctgaagg
     1861 gcagcgccga gggcagcatc agataccccg cctggaacgg catcatcagc caggaggtgc
     1921 tggactacct gagcagctac atcaacaggc ggatctgaac gcgtatttga tccgctagca
     1981 gatctgcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2041 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2101 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2161 agggggagga ttgggaagac aatagcaggc atgctgggga cacgtgcgga ccgagcggcc
     2221 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     2281 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     2341 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     2401 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     2461 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2521 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2581 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2641 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2701 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2761 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     2821 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2881 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2941 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3001 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3061 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     3121 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3181 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3241 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3301 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3361 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3421 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3481 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     3541 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3601 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3661 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3721 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3781 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3841 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3901 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3961 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4021 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4081 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4141 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4201 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4261 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4321 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4381 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4441 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     4501 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4561 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4621 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4681 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4741 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4801 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4861 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4921 ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
