LOCUS       sequence-75168-e        4972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Can be used to generate AAV virus that will express FlpE recombinse 
            gene in neurons from the synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4972)
  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 4972)
  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)-FlpE-bGHpA
FEATURES             Location/Qualifiers
     source          1..4972
                     /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)"
     CDS             join(658..663,664..684,685..1956)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPe
                     /note="FLPe is a fusion between the SV40 nuclear 
                     localization signal and a thermostable version of the 
                     Saccharomyces cerevisiae site-specific recombinase FLP. 
                     Recombination occurs at FRT sequences (Buchholz et al., 
                     1998)."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     primer_bind     complement(1999..2016)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2005..2212
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2235..2375
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2235..2364
                     /label=AAV2 ITR
     rep_origin      2450..2905
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2537..2556)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2747..2768
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2922..2941)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3041..3063
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3101..3119)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3187..3291
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3292..3360,3361..4152)
                     /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(3510..3529)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4323..4911
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4812..4831
                     /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 cttcaggtcg acccaccatg
      661 gctcccaaga agaagaggaa ggtgatgagt caatttgata tattatgtaa aacaccacct
      721 aaggtcctgg ttcgtcagtt tgtggaaagg tttgaaagac cttcagggga aaaaatagca
      781 tcatgtgctg ctgaactaac ctatttatgt tggatgatta ctcataacgg aacagcaatc
      841 aagagagcca cattcatgag ctataatact atcataagca attcgctgag tttcgatatt
      901 gtcaacaaat cactccagtt taaatacaag acgcaaaaag caacaattct ggaagcctca
      961 ttaaagaaat taattcctgc ttgggaattt acaattattc cttacaatgg acaaaaacat
     1021 caatctgata tcactgatat tgtaagtagt ttgcaattac agttcgaatc atcggaagaa
     1081 gcagataagg gaaatagcca cagtaaaaaa atgcttaaag cacttctaag tgagggtgaa
     1141 agcatctggg agatcactga gaaaatacta aattcgtttg agtatacctc gagatttaca
     1201 aaaacaaaaa ctttatacca attcctcttc ctagctactt tcatcaattg tggaagattc
     1261 agcgatatta agaacgttga tccgaaatca tttaaattag tccaaaataa gtatctggga
     1321 gtaataatcc agtgtttagt gacagagaca aagacaagcg ttagtaggca catatacttc
     1381 tttagcgcaa ggggtaggat cgatccactt gtatatttgg atgaattttt gaggaattct
     1441 gaaccagtcc taaaacgagt aaataggacc ggcaattctt caagcaacaa acaggaatac
     1501 caattattaa aagataactt agtcagatcg tacaacaagg ctttgaagaa aaatgcgcct
     1561 tatccaatct ttgctataaa gaatggccca aaatctcaca ttggaagaca tttgatgacc
     1621 tcatttctgt caatgaaggg cctaacggag ttgactaatg ttgtgggaaa ttggagcgat
     1681 aagcgtgctt ctgccgtggc caggacaacg tatactcatc agataacagc aatacctgat
     1741 cactacttcg cactagtttc tcggtactat gcatatgatc caatatcaaa ggaaatgata
     1801 gcattgaagg atgagactaa tccaattgag gagtggcagc atatagaaca gctaaagggt
     1861 agtgctgaag gaagcatacg ataccccgca tggaatggga taatatcaca ggaggtacta
     1921 gactaccttt catcctacat aaatagacgc atataatgat ctagagtcga gttaattaag
     1981 aattccgccc cagatctgcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2041 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2101 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2161 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gacacgtgcg
     2221 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2281 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     2341 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     2401 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     2461 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2521 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2581 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2641 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     2701 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2761 caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg
     2821 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2881 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     2941 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3001 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3061 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3121 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3181 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3241 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3301 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3361 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3421 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3481 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3541 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3601 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3661 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3721 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3781 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3841 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3901 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3961 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4021 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4081 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4141 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4201 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4261 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4321 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4381 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4441 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     4501 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4561 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4621 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4681 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4741 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4801 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4861 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4921 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
