LOCUS       sequence-470291-        6468 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2047m 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 6468)
  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 6468)
  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: AiP1191 - pAAV-AiE2047m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1266..1275
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1272..1277,1278..1298,1299..2570)
                     /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    2595..3183
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2648..2668)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3066..3077)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3215..3691
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3414..3433)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3731..3871
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3731..3860
                     /label=AAV2 ITR
     rep_origin      3946..4401
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4033..4052)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4243..4264
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4418..4437)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4537..4559
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4597..4615)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4683..4787
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4788..4856,4857..5648)
                     /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(5006..5025)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5819..6407
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6308..6327
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgacctca
       61 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc
      121 cgcacgcgtg tgttatcctt cttgaccttc tccataagtg acaagtggct cgcctaaagt
      181 cactcattct caacgtgaaa ggggaattga ggatccttag tgatttcatg attcccaaat
      241 tatgtctaaa caaagacaat gctatctcaa tgtataagat gactactggc ctatatttac
      301 ctgacaagaa tctattcctc cttcttcata tcctgcacat atcatacttt cagtgatgtt
      361 gtattctggc agctgttgct ggcacttttc attggatata agaggaacat cagcttcttt
      421 cagcacatca acagtagaac ctgcttgaaa tgggaaatgc agacagccag tcagtaatga
      481 ttaacaggca tcctcgaatg aaacgccatc catctgattt atcccagcat tttgctctca
      541 ttatcaagta ctatgtggat gtataatgga actaactacc aaccttgcac tttcaagtac
      601 ataataaacc catctgtgca ctttggcaaa tggagagcat ttctttcttc ctttcctaag
      661 gggtagtaag aattgtggat tggactaggg tggtctttta ggcattctca gtatttttta
      721 aagagttaca gtgttgctgg tagaaaggaa gtctgccagc acaaagtgtt cattcatgga
      781 aatcagcaca gatttggata tccagaagag cttttcatcc agcacctgag gaggtgctcc
      841 ctcctgtata aaggcattca gctcctttgt ggttctgtgt gtgcgagagc aggtgcacgc
      901 tgaacagggg acaaagccag ctgaactgtg ctctactctt ggggcaatat gcttgccatc
      961 ccaggcaggt gaagagaaag gaagcatatt tcagcattgt gtgtttgcat ttgttttatc
     1021 acaaatcact cactgcaagg atgaacctct ccttactaaa ctactgttcc caagaagctt
     1081 gtgaagcaag caggtgttag taccttcaga aaaggaacat aagcatttat tcatgattag
     1141 caattctctt tgatgtcaca aaagggatag aaacttaagt gtaacttatt atatctgtca
     1201 ggagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg
     1261 gcgtggccac catggctcct aagaagaaga ggaaggtgat gagccagttc gacatcctgt
     1321 gcaagacccc ccccaaggtg ctggtgcggc agttcgtgga gagattcgag aggcccagcg
     1381 gcgagaagat cgccagctgt gccgccgagc tgacctacct gtgctggatg atcacccaca
     1441 acggcaccgc catcaagagg gccaccttca tgagctacaa caccatcatc agcaacagcc
     1501 tgagcttcga catcgtgaac aagagcctgc agttcaagta caagacccag aaggccacca
     1561 tcctggaggc cagcctgaag aagctgatcc ccgcctggga gttcaccatc atcccttaca
     1621 acggccagaa gcaccagagc gacatcaccg acatcgtgtc cagcctgcag ctgcagttcg
     1681 agagcagcga ggaggccgac aagggcaaca gccacagcaa gaagatgctg aaggccctgc
     1741 tgtccgaggg cgagagcatc tgggagatca ccgagaagat cctgaacagc ttcgagtaca
     1801 ccagcaggtt caccaagacc aagaccctgt accagttcct gttcctggcc acattcatca
     1861 actgcggcag gttcagcgac atcaagaacg tggaccccaa gagcttcaag ctggtgcaga
     1921 acaagtacct gggcgtgatc attcagtgcc tggtgaccga gaccaagaca agcgtgtcca
     1981 ggcacatcta ctttttcagc gccagaggca ggatcgaccc cctggtgtac ctggacgagt
     2041 tcctgaggaa cagcgagccc gtgctgaaga gagtgaacag gaccggcaac agcagcagca
     2101 acaagcagga gtaccagctg ctgaaggaca acctggtgcg cagctacaac aaggccctga
     2161 agaagaacgc cccctacccc atcttcgcta tcaagaacgg ccctaagagc cacatcggca
     2221 ggcacctgat gaccagcttt ctgagcatga agggcctgac cgagctgaca aacgtggtgg
     2281 gcaactggag cgacaagagg gcctccgccg tggccaggac cacctacacc caccagatca
     2341 ccgccatccc cgaccactac ttcgccctgg tgtccaggta ctacgcctac gaccccatca
     2401 gcaaggagat gatcgccctg aaggacgaga ccaaccccat cgaggagtgg cagcacatcg
     2461 agcagctgaa gggcagcgcc gagggcagca tcagataccc cgcctggaac ggcatcatca
     2521 gccaggaggt gctggactac ctgagcagct acatcaacag gcggatctga gaattcgata
     2581 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     2641 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     2701 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     2761 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     2821 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     2881 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     2941 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     3001 ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     3061 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     3121 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     3181 cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc
     3241 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     3301 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     3361 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     3421 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     3481 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     3541 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     3601 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     3661 ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc
     3721 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     3781 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     3841 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     3901 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     3961 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4021 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4081 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4141 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     4201 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4261 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     4321 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4381 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     4441 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4501 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4561 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     4621 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4681 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4741 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4801 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4861 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4921 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4981 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5041 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5101 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5161 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5221 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5281 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5341 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5401 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5461 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5521 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5581 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5641 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5701 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5761 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5821 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5881 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5941 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6001 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6061 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6121 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6181 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6241 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6301 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6361 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6421 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
