LOCUS       sequence-325786-        5994 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing FlpO AAV Virus. Alias: AiP1037 - 
            pAAV-AiE2013m-minBG-FlpO-WPRE-HGHpA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5994)
  AUTHORS   Graybuck LT, Daigle TL, Sedeno-Cortes AE, Walker M, Kalmbach B, Lenz
            GH, Morin E, Nguyen TN, Garren E, Bendrick JL, Kim TK, Zhou T, 
            Mortrud M, Yao S, Siverts LA, Larsen R, Gore BB, Szelenyi ER, Trader
            C, Balaram P, van Velthoven CTJ, Chiang M, Mich JK, Dee N, Goldy J, 
            Cetin AH, Smith K, Way SW, Esposito L, Yao Z, Gradinaru V, Sunkin 
            SM, Lein E, Levi BP, Ting JT, Zeng H, Tasic B
  TITLE     Enhancer viruses for combinatorial cell-subclass-specific labeling.
  JOURNAL   Neuron. 2021 Mar 23. pii: S0896-6273(21)00159-8. doi: 
            10.1016/j.neuron.2021.03.011.
  PUBMED    33789083
REFERENCE   2  (bases 1 to 5994)
  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: "Neuron. 
            2021 Mar 23. pii: S0896-6273(21)00159-8. doi: 
            10.1016/j.neuron.2021.03.011."
COMMENT     Sequence Label: AiP1037-pAAV-mscRE13-minBGpromoter-FlpO-WPRE-h...
FEATURES             Location/Qualifiers
     source          1..5994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      792..801
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(798..803,804..824,825..2096)
                     /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    2121..2709
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2174..2194)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2592..2603)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2741..3217
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2940..2959)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3257..3397
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3257..3386
                     /label=AAV2 ITR
     rep_origin      3472..3927
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3559..3578)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3769..3790
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3944..3963)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4063..4085
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4123..4141)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4209..4313
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4314..4382,4383..5174)
                     /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(4532..4551)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5345..5933
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5834..5853
                     /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 gcgtgtccca taggcagttt gtggctgagt gctggtccag
      181 gggtgaggag gtgggctatg ttactgaggg tctctgggtg ttaggaaaac agggcccagg
      241 agtctggctg ctcgtatgct ggcccaggct cttgtttttc ttgagctgac ttgctggaga
      301 agtgagctaa gtcagaaaca aaatgccaca ttgcacgccc actgaagtct gggctcaagg
      361 gaaagaagag agattgccag agcgttagct gttcccaatc cactcctgga ccttaagctg
      421 tcttgaacag agttgccaat cagcttggta gggactggcc tttgaggagg ggagggggtg
      481 taggcagggg agggggagag aagggagcag tctgcgctcc atcttaatta cctcatcaga
      541 aacagctccc ttcccgcaaa gctctggtgt cttctacaag agggtgagtc tttggcttta
      601 catgtgaact tgtgccattt gcctgcgtat ataaacatga agggtcgtct gggttcagag
      661 ctgaaatctt tcacttgtga cttagctggg aaattcttgg caagatcaga atgcagtggt
      721 aaggtctgag ctcgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg
      781 cttctggcgt ggccaccatg gctcctaaga agaagaggaa ggtgatgagc cagttcgaca
      841 tcctgtgcaa gacccccccc aaggtgctgg tgcggcagtt cgtggagaga ttcgagaggc
      901 ccagcggcga gaagatcgcc agctgtgccg ccgagctgac ctacctgtgc tggatgatca
      961 cccacaacgg caccgccatc aagagggcca ccttcatgag ctacaacacc atcatcagca
     1021 acagcctgag cttcgacatc gtgaacaaga gcctgcagtt caagtacaag acccagaagg
     1081 ccaccatcct ggaggccagc ctgaagaagc tgatccccgc ctgggagttc accatcatcc
     1141 cttacaacgg ccagaagcac cagagcgaca tcaccgacat cgtgtccagc ctgcagctgc
     1201 agttcgagag cagcgaggag gccgacaagg gcaacagcca cagcaagaag atgctgaagg
     1261 ccctgctgtc cgagggcgag agcatctggg agatcaccga gaagatcctg aacagcttcg
     1321 agtacaccag caggttcacc aagaccaaga ccctgtacca gttcctgttc ctggccacat
     1381 tcatcaactg cggcaggttc agcgacatca agaacgtgga ccccaagagc ttcaagctgg
     1441 tgcagaacaa gtacctgggc gtgatcattc agtgcctggt gaccgagacc aagacaagcg
     1501 tgtccaggca catctacttt ttcagcgcca gaggcaggat cgaccccctg gtgtacctgg
     1561 acgagttcct gaggaacagc gagcccgtgc tgaagagagt gaacaggacc ggcaacagca
     1621 gcagcaacaa gcaggagtac cagctgctga aggacaacct ggtgcgcagc tacaacaagg
     1681 ccctgaagaa gaacgccccc taccccatct tcgctatcaa gaacggccct aagagccaca
     1741 tcggcaggca cctgatgacc agctttctga gcatgaaggg cctgaccgag ctgacaaacg
     1801 tggtgggcaa ctggagcgac aagagggcct ccgccgtggc caggaccacc tacacccacc
     1861 agatcaccgc catccccgac cactacttcg ccctggtgtc caggtactac gcctacgacc
     1921 ccatcagcaa ggagatgatc gccctgaagg acgagaccaa ccccatcgag gagtggcagc
     1981 acatcgagca gctgaagggc agcgccgagg gcagcatcag ataccccgcc tggaacggca
     2041 tcatcagcca ggaggtgctg gactacctga gcagctacat caacaggcgg atctgagaat
     2101 tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     2161 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2221 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     2281 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     2341 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     2401 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     2461 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     2521 cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct
     2581 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     2641 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     2701 cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc
     2761 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
     2821 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
     2881 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
     2941 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
     3001 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat
     3061 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag
     3121 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
     3181 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg
     3241 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3301 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3361 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     3421 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     3481 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3541 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3601 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3661 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     3721 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3781 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     3841 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3901 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     3961 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4021 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4081 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     4141 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4201 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4261 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4321 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4381 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4441 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4501 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4561 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4621 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4681 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4741 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4801 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4861 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4921 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4981 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5041 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5101 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5161 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5221 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5281 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5341 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5401 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5461 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5521 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5581 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5641 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5701 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5761 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5821 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5881 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5941 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
