LOCUS       sequence-343150-        5495 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  also known as: pAAV-synP-FLPo. AAV virus for expressing FLPo 
            recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5495)
  AUTHORS   Jin L, Sullivan HA, Zhu M, Lavin TK, Matsuyama M, Fu X, Lea NE, Xu 
            R, Hou Y, Rutigliani L, Pruner M, Babcock KR, Ip JPK, Hu M, Daigle 
            TL, Zeng H, Sur M, Feng G, Wickersham IR
  TITLE     Long-term labeling and imaging of synaptically connected neuronal 
            networks in vivo using double-deletion-mutant rabies viruses.
  JOURNAL   Nat Neurosci. 2024 Jan 11. doi: 10.1038/s41593-023-01545-8.
  PUBMED    38212587
REFERENCE   2  (bases 1 to 5495)
  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: "Nat 
            Neurosci. 2024 Jan 11. doi: 10.1038/s41593-023-01545-8."
COMMENT     Sequence Label: pAAV-syn-Flpo
FEATURES             Location/Qualifiers
     source          1..5495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      460..469
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(466..471,472..492,493..1764)
                     /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"
     misc_feature    1773..2361
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1826..1846)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2244..2255)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2369..2593
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2595..2735
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2595..2724
                     /label=AAV2 ITR
     rep_origin      2810..3265
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2897..2916)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3107..3128
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3282..3301)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3401..3423
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3461..3479)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3547..3651
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3652..3720,3721..4512)
                     /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(3870..3889)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4683..5271
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5172..5191
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5333..5462
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagtc gaattcgccg ccaccatggc tcctaagaag
      481 aagaggaagg tgatgagcca gttcgacatc ctgtgcaaga ccccccccaa ggtgctggtg
      541 cggcagttcg tggagagatt cgagaggccc agcggcgaga agatcgccag ctgtgccgcc
      601 gagctgacct acctgtgctg gatgatcacc cacaacggca ccgccatcaa gagggccacc
      661 ttcatgagct acaacaccat catcagcaac agcctgagct tcgacatcgt gaacaagagc
      721 ctgcagttca agtacaagac ccagaaggcc accatcctgg aggccagcct gaagaagctg
      781 atccccgcct gggagttcac catcatccct tacaacggcc agaagcacca gagcgacatc
      841 accgacatcg tgtccagcct gcagctgcag ttcgagagca gcgaggaggc cgacaagggc
      901 aacagccaca gcaagaagat gctgaaggcc ctgctgtccg agggcgagag catctgggag
      961 atcaccgaga agatcctgaa cagcttcgag tacaccagca ggttcaccaa gaccaagacc
     1021 ctgtaccagt tcctgttcct ggccacattc atcaactgcg gcaggttcag cgacatcaag
     1081 aacgtggacc ccaagagctt caagctggtg cagaacaagt acctgggcgt gatcattcag
     1141 tgcctggtga ccgagaccaa gacaagcgtg tccaggcaca tctacttttt cagcgccaga
     1201 ggcaggatcg accccctggt gtacctggac gagttcctga ggaacagcga gcccgtgctg
     1261 aagagagtga acaggaccgg caacagcagc agcaacaagc aggagtacca gctgctgaag
     1321 gacaacctgg tgcgcagcta caacaaggcc ctgaagaaga acgcccccta ccccatcttc
     1381 gctatcaaga acggccctaa gagccacatc ggcaggcacc tgatgaccag ctttctgagc
     1441 atgaagggcc tgaccgagct gacaaacgtg gtgggcaact ggagcgacaa gagggcctcc
     1501 gccgtggcca ggaccaccta cacccaccag atcaccgcca tccccgacca ctacttcgcc
     1561 ctggtgtcca ggtactacgc ctacgacccc atcagcaagg agatgatcgc cctgaaggac
     1621 gagaccaacc ccatcgagga gtggcagcac atcgagcagc tgaagggcag cgccgagggc
     1681 agcatcagat accccgcctg gaacggcatc atcagccagg aggtgctgga ctacctgagc
     1741 agctacatca acaggcggat ctgataatcc ggaatcaacc tctggattac aaaatttgtg
     1801 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     1861 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     1921 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     1981 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     2041 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     2101 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     2161 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     2221 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     2281 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     2341 ccctttgggc cgcctccccg cctggatcct gtgccttcta gttgccagcc atctgttgtt
     2401 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2461 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2521 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2581 gtgggctcta tggcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2641 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     2701 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     2761 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     2821 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2881 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2941 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3001 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     3061 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3121 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     3181 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3241 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     3301 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3361 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3421 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3481 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3541 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3601 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3661 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3721 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3781 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3841 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3901 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3961 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4021 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4081 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4141 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4201 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4261 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4321 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4381 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4441 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4501 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4561 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4621 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4681 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4741 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4801 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4861 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4921 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4981 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5041 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5101 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5161 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5221 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5281 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg
     5341 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc
     5401 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
     5461 ctgtcgacgc tagactgcag agggccctgc gtatg
//
