LOCUS       sequence-305863-        4949 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 4949)
  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 4949)
  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..4949
                     /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        166..613
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      643..652
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(649..654,655..675,676..1947)
                     /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(1976..1993)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1982..2189
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2212..2352
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2212..2341
                     /label=AAV2 ITR
     rep_origin      2427..2882
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2514..2533)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2724..2745
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2899..2918)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3018..3040
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3078..3096)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3164..3268
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3269..3337,3338..4129)
                     /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(3487..3506)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4300..4888
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4789..4808
                     /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 gcgtctgcag agggccctgc gtatgagtgc aagtgggttt
      181 taggaccagg atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga
      241 caagcaccca acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa
      301 caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc
      361 cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc
      421 cggtcccccg caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc
      481 agccggaccg caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg
      541 cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg
      601 cctgagagcg cagacgcgtg gacggtacca agcttacgcg tagccaccat ggctcctaag
      661 aagaagagga aggtgatgag ccagttcgac atcctgtgca agaccccccc caaggtgctg
      721 gtgcggcagt tcgtggagag attcgagagg cccagcggcg agaagatcgc cagctgtgcc
      781 gccgagctga cctacctgtg ctggatgatc acccacaacg gcaccgccat caagagggcc
      841 accttcatga gctacaacac catcatcagc aacagcctga gcttcgacat cgtgaacaag
      901 agcctgcagt tcaagtacaa gacccagaag gccaccatcc tggaggccag cctgaagaag
      961 ctgatccccg cctgggagtt caccatcatc ccttacaacg gccagaagca ccagagcgac
     1021 atcaccgaca tcgtgtccag cctgcagctg cagttcgaga gcagcgagga ggccgacaag
     1081 ggcaacagcc acagcaagaa gatgctgaag gccctgctgt ccgagggcga gagcatctgg
     1141 gagatcaccg agaagatcct gaacagcttc gagtacacca gcaggttcac caagaccaag
     1201 accctgtacc agttcctgtt cctggccaca ttcatcaact gcggcaggtt cagcgacatc
     1261 aagaacgtgg accccaagag cttcaagctg gtgcagaaca agtacctggg cgtgatcatt
     1321 cagtgcctgg tgaccgagac caagacaagc gtgtccaggc acatctactt tttcagcgcc
     1381 agaggcagga tcgaccccct ggtgtacctg gacgagttcc tgaggaacag cgagcccgtg
     1441 ctgaagagag tgaacaggac cggcaacagc agcagcaaca agcaggagta ccagctgctg
     1501 aaggacaacc tggtgcgcag ctacaacaag gccctgaaga agaacgcccc ctaccccatc
     1561 ttcgctatca agaacggccc taagagccac atcggcaggc acctgatgac cagctttctg
     1621 agcatgaagg gcctgaccga gctgacaaac gtggtgggca actggagcga caagagggcc
     1681 tccgccgtgg ccaggaccac ctacacccac cagatcaccg ccatccccga ccactacttc
     1741 gccctggtgt ccaggtacta cgcctacgac cccatcagca aggagatgat cgccctgaag
     1801 gacgagacca accccatcga ggagtggcag cacatcgagc agctgaaggg cagcgccgag
     1861 ggcagcatca gataccccgc ctggaacggc atcatcagcc aggaggtgct ggactacctg
     1921 agcagctaca tcaacaggcg gatctgaacg cgtatttgat ccgctagcag atctgcctcg
     1981 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2041 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2101 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2161 tgggaagaca atagcaggca tgctggggac acgtgcggac cgagcggccg caggaacccc
     2221 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     2281 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     2341 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     2401 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     2461 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     2521 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2581 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2641 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2701 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     2761 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     2821 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     2881 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     2941 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3001 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3061 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     3121 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3181 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3241 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3301 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3361 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3421 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3481 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3541 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3601 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3661 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3721 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3781 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3841 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3901 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3961 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4021 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4081 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4141 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4201 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4261 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4321 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4381 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4441 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4501 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4561 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4621 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4681 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4741 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4801 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4861 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4921 ggccttttgc tggccttttg ctcacatgt
//
