LOCUS       sequence-461957-        5769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2051m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific populations of brain cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5769)
  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 5769)
  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: AiP1211 - pAAV-AiE2051m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..5769
                     /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      567..576
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(573..578,579..599,600..1871)
                     /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    1896..2484
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1949..1969)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2367..2378)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2516..2992
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2715..2734)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3032..3172
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3032..3161
                     /label=AAV2 ITR
     rep_origin      3247..3702
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3334..3353)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3544..3565
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3719..3738)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3838..3860
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3898..3916)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3984..4088
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4089..4157,4158..4949)
                     /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(4307..4326)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5120..5708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5609..5628
                     /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 gcgtcctttt agaaacactc cctttaccat cttcaaacat
      181 gtttttagct gagaactctc ttccaacccc tgtttcttaa ctctgagtgc cgtatcactt
      241 catattctgt gtcaggggaa aagaaatcag cacctcccac aacgtcaact gtgtccttta
      301 gagtaagcat tccaaagtct atattctgag tctaatctct cttggaaagg tcagcctttt
      361 cttccaaata ccaagtggac atctttatgc acaggccttc aaggatattc aagtcagtgt
      421 gtcctacgga gaaaactcct tttaaaaccc gaatcctgtg gtttccccaa gatgattatt
      481 gggtggtcac tgcagcctta aggagctcgg gctgggcata aaagtcaggg cagagccatc
      541 tattgcttac atttgcttct ggcgtggcca ccatggctcc taagaagaag aggaaggtga
      601 tgagccagtt cgacatcctg tgcaagaccc cccccaaggt gctggtgcgg cagttcgtgg
      661 agagattcga gaggcccagc ggcgagaaga tcgccagctg tgccgccgag ctgacctacc
      721 tgtgctggat gatcacccac aacggcaccg ccatcaagag ggccaccttc atgagctaca
      781 acaccatcat cagcaacagc ctgagcttcg acatcgtgaa caagagcctg cagttcaagt
      841 acaagaccca gaaggccacc atcctggagg ccagcctgaa gaagctgatc cccgcctggg
      901 agttcaccat catcccttac aacggccaga agcaccagag cgacatcacc gacatcgtgt
      961 ccagcctgca gctgcagttc gagagcagcg aggaggccga caagggcaac agccacagca
     1021 agaagatgct gaaggccctg ctgtccgagg gcgagagcat ctgggagatc accgagaaga
     1081 tcctgaacag cttcgagtac accagcaggt tcaccaagac caagaccctg taccagttcc
     1141 tgttcctggc cacattcatc aactgcggca ggttcagcga catcaagaac gtggacccca
     1201 agagcttcaa gctggtgcag aacaagtacc tgggcgtgat cattcagtgc ctggtgaccg
     1261 agaccaagac aagcgtgtcc aggcacatct actttttcag cgccagaggc aggatcgacc
     1321 ccctggtgta cctggacgag ttcctgagga acagcgagcc cgtgctgaag agagtgaaca
     1381 ggaccggcaa cagcagcagc aacaagcagg agtaccagct gctgaaggac aacctggtgc
     1441 gcagctacaa caaggccctg aagaagaacg ccccctaccc catcttcgct atcaagaacg
     1501 gccctaagag ccacatcggc aggcacctga tgaccagctt tctgagcatg aagggcctga
     1561 ccgagctgac aaacgtggtg ggcaactgga gcgacaagag ggcctccgcc gtggccagga
     1621 ccacctacac ccaccagatc accgccatcc ccgaccacta cttcgccctg gtgtccaggt
     1681 actacgccta cgaccccatc agcaaggaga tgatcgccct gaaggacgag accaacccca
     1741 tcgaggagtg gcagcacatc gagcagctga agggcagcgc cgagggcagc atcagatacc
     1801 ccgcctggaa cggcatcatc agccaggagg tgctggacta cctgagcagc tacatcaaca
     1861 ggcggatctg agaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     1921 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     1981 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     2041 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     2101 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     2161 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     2221 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     2281 tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc
     2341 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     2401 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     2461 tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg
     2521 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     2581 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     2641 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     2701 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     2761 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     2821 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     2881 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     2941 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
     3001 gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     3061 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     3121 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     3181 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     3241 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3301 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     3361 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3421 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     3481 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3541 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     3601 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     3661 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     3721 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     3781 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     3841 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     3901 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     3961 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4021 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4081 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4141 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4201 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4261 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4321 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4381 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4441 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4501 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4561 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4621 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4681 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4741 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4801 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4861 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4921 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4981 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5041 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5101 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5161 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5221 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5281 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5341 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5401 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5461 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5521 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5581 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5641 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5701 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5761 ctcacatgt
//
