LOCUS       sequence-446285-        5191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FlpO recombinase expression from the DLX2.0 enhancer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5191)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle TL, Dwivedi D, Way SW, 
            Oster A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya
            X, Roth JR, Alexander JR, Allen S, Amster A, Arbuckle J, Ayala A, 
            Baker PM, Bakken TE, Barcelli T, Barta S, Bendrick J, Bertagnolli D,
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chance RK, Chavan S, Clark M, 
            Colbert K, Collman F, Daniel S, Departee M, DiValentin P, Donadio N,
            Dotson N, Egdorf T, Fliss T, Gabitto M, Garcia J, Gary A, Gasperini 
            M, Gloe J, Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, 
            Halterman C, Haradon Z, Hastings SD, Helback O, Ho W, Hockemeyer D, 
            Huang C, Huff S, Hunker A, Johansen N, Jones D, Juneau Z, Kalmbach 
            B, Kannan M, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, Lee AY, 
            Leibly M, Lenz GH, Li S, Liang E, Lusk N, Madigan Z, Malloy J, 
            Malone J, McCue R, Melchor J, Mollenkopf T, Moosman S, Morin E, 
            Newman D, Ng L, Ngo K, Omstead V, Otto S, Oyama A, Pena N, Pham T, 
            Phillips E, Pom CA, Potekhina L, Ransford S, Ray PL, Rette D, 
            Reynoldson C, Rimorin C, Rocha D, Ruiz A, Sanchez REA, Sawyer L, 
            Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shepard N, Shulga L, 
            Sigler AR, Siverts L, Soliman S, Somasundaram S, Staats B, Stewart 
            K, Szelenyi E, Tieu M, Trader C, Tran A, van Velthoven CTJ, Walker 
            M, Wang Y, Weed N, Wirthlin M, Wood T, Wynalda B, Yao Z, Zhou T, 
            Ariza J, Dee N, Reding M, Ronellenfitch K, Mufti S, Sunkin SM, Smith
            KA, Esposito L, Waters J, Thyagarajan B, Yao S, Lein ES, Zeng H, 
            Levi BP, Ngai J, Ting JT, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   Cell. 2025 May 16:S0092-8674(25)00513-6. doi: 
            10.1016/j.cell.2025.05.002.
  PUBMED    40403729
REFERENCE   2  (bases 1 to 5191)
  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: "Cell. 2025 
            May 16:S0092-8674(25)00513-6. doi: 10.1016/j.cell.2025.05.002."
COMMENT     Sequence Label: AiP14532-pAAV-DLX2.0-minBG-FlpO-WPRE3-BGHpA (A...
FEATURES             Location/Qualifiers
     source          1..5191
                     /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      624..633
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(630..635,636..656,657..1928)
                     /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(2018..2038)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2225..2336
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2454..2594
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2454..2583
                     /label=AAV2 ITR
     rep_origin      2669..3124
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2756..2775)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2966..2987
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3141..3160)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3260..3282
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3320..3338)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3406..3510
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3511..3579,3580..4371)
                     /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(3729..3748)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4542..5130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5031..5050
                     /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 gcgtggtacc ctaaataaag atggcttttt agtattaaaa
      181 gtggaagaaa attacaggta attatctttg acggtaaaaa cgctgtaatc agcgggctac
      241 atgaaaaatt actctaatta tggctgcatt taagagaatg gctaaataaa gatggctttt
      301 tagtattaaa agtggaagaa aattacaggt aattatcttt gacggtaaaa acgctgtaat
      361 cagcgggcta catgaaaaat tactctaatt atggctgcat ttaagagaat ggctaaataa
      421 agatggcttt ttagtattaa aagtggaaga aaattacagg taattatctt tgacggtaaa
      481 aacgctgtaa tcagcgggct acatgaaaaa ttactctaat tatggctgca tttaagagaa
      541 tggagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct
      601 gggatccaga tctttcgaag ctagccacca tggctcctaa gaagaagagg aaggtgatga
      661 gccagttcga catcctgtgc aagacccccc ccaaggtgct ggtgcggcag ttcgtggaga
      721 gattcgagag gcccagcggc gagaagatcg ccagctgtgc cgccgagctg acctacctgt
      781 gctggatgat cacccacaac ggcaccgcca tcaagagggc caccttcatg agctacaaca
      841 ccatcatcag caacagcctg agcttcgaca tcgtgaacaa gagcctgcag ttcaagtaca
      901 agacccagaa ggccaccatc ctggaggcca gcctgaagaa gctgatcccc gcctgggagt
      961 tcaccatcat cccttacaac ggccagaagc accagagcga catcaccgac atcgtgtcca
     1021 gcctgcagct gcagttcgag agcagcgagg aggccgacaa gggcaacagc cacagcaaga
     1081 agatgctgaa ggccctgctg tccgagggcg agagcatctg ggagatcacc gagaagatcc
     1141 tgaacagctt cgagtacacc agcaggttca ccaagaccaa gaccctgtac cagttcctgt
     1201 tcctggccac attcatcaac tgcggcaggt tcagcgacat caagaacgtg gaccccaaga
     1261 gcttcaagct ggtgcagaac aagtacctgg gcgtgatcat tcagtgcctg gtgaccgaga
     1321 ccaagacaag cgtgtccagg cacatctact ttttcagcgc cagaggcagg atcgaccccc
     1381 tggtgtacct ggacgagttc ctgaggaaca gcgagcccgt gctgaagaga gtgaacagga
     1441 ccggcaacag cagcagcaac aagcaggagt accagctgct gaaggacaac ctggtgcgca
     1501 gctacaacaa ggccctgaag aagaacgccc cctaccccat cttcgctatc aagaacggcc
     1561 ctaagagcca catcggcagg cacctgatga ccagctttct gagcatgaag ggcctgaccg
     1621 agctgacaaa cgtggtgggc aactggagcg acaagagggc ctccgccgtg gccaggacca
     1681 cctacaccca ccagatcacc gccatccccg accactactt cgccctggtg tccaggtact
     1741 acgcctacga ccccatcagc aaggagatga tcgccctgaa ggacgagacc aaccccatcg
     1801 aggagtggca gcacatcgag cagctgaagg gcagcgccga gggcagcatc agataccccg
     1861 cctggaacgg catcatcagc caggaggtgc tggactacct gagcagctac atcaacaggc
     1921 ggatctgagt cgacggcgcg ccgcggccgc gaattcgata tcataatcaa cctctggatt
     1981 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2041 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2101 cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg
     2161 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct cgagagatct
     2221 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2281 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2341 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2401 gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc
     2461 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     2521 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     2581 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2641 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     2701 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     2761 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2821 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2881 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2941 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3001 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     3061 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     3121 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     3181 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3241 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3301 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3361 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3421 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3481 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3541 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3601 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3661 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3721 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3781 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3841 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3901 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3961 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4021 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4081 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4141 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4201 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4261 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4321 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4381 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4441 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4501 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4561 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4621 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4681 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4741 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4801 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4861 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4921 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4981 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5041 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5101 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5161 ctggcctttt gctggccttt tgctcacatg t
//
