LOCUS       sequence-245621-        6922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated Cre-dependent expression of FLPo recombinase under EF1a
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6922)
  AUTHORS   Trouche S, Koren V, Doig NM, Ellender TJ, El-Gaby M, 
            Lopes-Dos-Santos V, Reeve HM, Perestenko PV, Garas FN, Magill PJ, 
            Sharott A, Dupret D
  TITLE     A Hippocampus-Accumbens Tripartite Neuronal Motif Guides Appetitive 
            Memory in Space.
  JOURNAL   Cell. 2019 Mar 7;176(6):1393-1406.e16. doi: 
            10.1016/j.cell.2018.12.037. Epub 2019 Feb 14.
  PUBMED    30773318
REFERENCE   2  (bases 1 to 6922)
  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.cell.2018.12.037"; journalName: "Cell"; date: "2019-03-7-
            7"; volume: "176"; issue: "6"; pages: "1393-1406.e16"
COMMENT     Sequence Label: pAAV-EF1a-DIO-FLPo-Myc
FEATURES             Location/Qualifiers
     source          1..6922
                     /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        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1440..1473)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1524..1557)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1569..1598)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(join(1599..2867,2868..2888,2889..2894))
                     /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"
     regulatory      2891..2900
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2907..2940
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2991..3024
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3049..3637
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3102..3122)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3520..3531)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3669..4145
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3868..3887)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4185..4325
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4185..4314
                     /label=AAV2 ITR
     rep_origin      4400..4855
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4487..4506)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4697..4718
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4872..4891)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4991..5013
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5051..5069)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5137..5241
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5242..5310,5311..6102)
                     /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(5460..5479)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6273..6861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6762..6781
                     /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa
     1441 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
     1561 gcgcctcaca agtcctcttc agaaatgagc ttttgctcga tccgcctgtt gatgtagctg
     1621 ctcaggtagt ccagcacctc ctggctgatg atgccgttcc aggcggggta tctgatgctg
     1681 ccctcggcgc tgcccttcag ctgctcgatg tgctgccact cctcgatggg gttggtctcg
     1741 tccttcaggg cgatcatctc cttgctgatg gggtcgtagg cgtagtacct ggacaccagg
     1801 gcgaagtagt ggtcggggat ggcggtgatc tggtgggtgt aggtggtcct ggccacggcg
     1861 gaggccctct tgtcgctcca gttgcccacc acgtttgtca gctcggtcag gcccttcatg
     1921 ctcagaaagc tggtcatcag gtgcctgccg atgtggctct tagggccgtt cttgatagcg
     1981 aagatggggt agggggcgtt cttcttcagg gccttgttgt agctgcgcac caggttgtcc
     2041 ttcagcagct ggtactcctg cttgttgctg ctgctgttgc cggtcctgtt cactctcttc
     2101 agcacgggct cgctgttcct caggaactcg tccaggtaca ccagggggtc gatcctgcct
     2161 ctggcgctga aaaagtagat gtgcctggac acgcttgtct tggtctcggt caccaggcac
     2221 tgaatgatca cgcccaggta cttgttctgc accagcttga agctcttggg gtccacgttc
     2281 ttgatgtcgc tgaacctgcc gcagttgatg aatgtggcca ggaacaggaa ctggtacagg
     2341 gtcttggtct tggtgaacct gctggtgtac tcgaagctgt tcaggatctt ctcggtgatc
     2401 tcccagatgc tctcgccctc ggacagcagg gccttcagca tcttcttgct gtggctgttg
     2461 cccttgtcgg cctcctcgct gctctcgaac tgcagctgca ggctggacac gatgtcggtg
     2521 atgtcgctct ggtgcttctg gccgttgtaa gggatgatgg tgaactccca ggcggggatc
     2581 agcttcttca ggctggcctc caggatggtg gccttctggg tcttgtactt gaactgcagg
     2641 ctcttgttca cgatgtcgaa gctcaggctg ttgctgatga tggtgttgta gctcatgaag
     2701 gtggccctct tgatggcggt gccgttgtgg gtgatcatcc agcacaggta ggtcagctcg
     2761 gcggcacagc tggcgatctt ctcgccgctg ggcctctcga atctctccac gaactgccgc
     2821 accagcacct tggggggggt cttgcacagg atgtcgaact ggctcatcac cttcctcttc
     2881 ttcttaggag ccatggtggc gctagcataa cttcgtataa agtatcctat acgaagttat
     2941 ttgccttaac ccagaaatta tcactgttat tctttagaat ggtgcaaaga ataacttcgt
     3001 ataatgtatg ctatacgaag ttatgaattc gatatcaagc ttatcgataa tcaacctctg
     3061 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3121 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3181 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     3241 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     3301 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     3361 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3421 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     3481 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3541 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3601 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag
     3661 agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     3721 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     3781 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     3841 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     3901 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     3961 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     4021 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     4081 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     4141 tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     4201 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     4261 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     4321 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4381 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4441 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     4501 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4561 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     4621 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4681 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     4741 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     4801 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     4861 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4921 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4981 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     5041 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5101 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5161 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5221 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5281 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5341 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5401 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5461 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5521 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5581 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5641 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5701 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5761 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5821 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5881 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5941 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6001 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6061 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6121 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6181 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6241 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6301 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6361 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6421 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6481 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6541 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6601 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6661 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6721 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6781 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6841 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6901 tgctggcctt ttgctcacat gt
//
