LOCUS       sequence-470297-        5752 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2056m 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 5752)
  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 5752)
  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: AiP1217 - pAAV-AiE2056m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..5752
                     /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      550..559
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(556..561,562..582,583..1854)
                     /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    1879..2467
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1932..1952)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2350..2361)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2499..2975
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2698..2717)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3015..3155
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3015..3144
                     /label=AAV2 ITR
     rep_origin      3230..3685
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3317..3336)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3527..3548
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3702..3721)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3821..3843
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3881..3899)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3967..4071
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4072..4140,4141..4932)
                     /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(4290..4309)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5103..5691
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5592..5611
                     /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 gcgtggtgaa tgatggatta ttttaaagtg aagtttcaag
      181 aaaacctgtt cagctgtttt tgagacatga ccagcagagt gagaaaatct attttattcc
      241 tctttcaaaa gttgcccttt tctaatgtca agcaagtgca gaacggatga actttggaag
      301 tgtcagtctt cgagtgttat gtaatttgtt ctgagtatga attagtctag ctgagccttt
      361 gagtgcatta aaaacaagga agatgagcta aacatgtccc tgaagggcaa aagcactgtt
      421 agtccgctca gctactttag caagccattt cttctagagt ggagcgttag tcttactgta
      481 ctgtggagct cgggctgggc ataaaagtca gggcagagcc atctattgct tacatttgct
      541 tctggcgtgg ccaccatggc tcctaagaag aagaggaagg tgatgagcca gttcgacatc
      601 ctgtgcaaga ccccccccaa ggtgctggtg cggcagttcg tggagagatt cgagaggccc
      661 agcggcgaga agatcgccag ctgtgccgcc gagctgacct acctgtgctg gatgatcacc
      721 cacaacggca ccgccatcaa gagggccacc ttcatgagct acaacaccat catcagcaac
      781 agcctgagct tcgacatcgt gaacaagagc ctgcagttca agtacaagac ccagaaggcc
      841 accatcctgg aggccagcct gaagaagctg atccccgcct gggagttcac catcatccct
      901 tacaacggcc agaagcacca gagcgacatc accgacatcg tgtccagcct gcagctgcag
      961 ttcgagagca gcgaggaggc cgacaagggc aacagccaca gcaagaagat gctgaaggcc
     1021 ctgctgtccg agggcgagag catctgggag atcaccgaga agatcctgaa cagcttcgag
     1081 tacaccagca ggttcaccaa gaccaagacc ctgtaccagt tcctgttcct ggccacattc
     1141 atcaactgcg gcaggttcag cgacatcaag aacgtggacc ccaagagctt caagctggtg
     1201 cagaacaagt acctgggcgt gatcattcag tgcctggtga ccgagaccaa gacaagcgtg
     1261 tccaggcaca tctacttttt cagcgccaga ggcaggatcg accccctggt gtacctggac
     1321 gagttcctga ggaacagcga gcccgtgctg aagagagtga acaggaccgg caacagcagc
     1381 agcaacaagc aggagtacca gctgctgaag gacaacctgg tgcgcagcta caacaaggcc
     1441 ctgaagaaga acgcccccta ccccatcttc gctatcaaga acggccctaa gagccacatc
     1501 ggcaggcacc tgatgaccag ctttctgagc atgaagggcc tgaccgagct gacaaacgtg
     1561 gtgggcaact ggagcgacaa gagggcctcc gccgtggcca ggaccaccta cacccaccag
     1621 atcaccgcca tccccgacca ctacttcgcc ctggtgtcca ggtactacgc ctacgacccc
     1681 atcagcaagg agatgatcgc cctgaaggac gagaccaacc ccatcgagga gtggcagcac
     1741 atcgagcagc tgaagggcag cgccgagggc agcatcagat accccgcctg gaacggcatc
     1801 atcagccagg aggtgctgga ctacctgagc agctacatca acaggcggat ctgagaattc
     1861 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     1921 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     1981 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     2041 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     2101 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     2161 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2221 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     2281 tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc
     2341 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     2401 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     2461 tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct
     2521 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     2581 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     2641 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     2701 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     2761 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     2821 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     2881 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     2941 tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg
     3001 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     3061 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     3121 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     3181 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     3241 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3301 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3361 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3421 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     3481 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3541 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     3601 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3661 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     3721 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3781 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3841 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3901 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3961 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4021 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4081 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4141 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4201 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4261 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     4321 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4381 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4441 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4501 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4561 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4621 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4681 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4741 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4801 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4861 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4921 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4981 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5041 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5101 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5161 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5221 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5281 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5341 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5401 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5461 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5521 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5581 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5641 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5701 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
