LOCUS       sequence-466206-        5945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2045m 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 5945)
  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 5945)
  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: AiP1186 - pAAV-AiE2045m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..5945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   84..213
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(275..863)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(355..374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1034..1825,1826..1894))
                     /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     1657..1676
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1895..1999)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2067..2085
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2123..2145)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2245..2264
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2281..2736)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2418..2439)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2630..2649
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2811..2951
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2822..2951
                     /label=AAV2 ITR
     polyA_signal    2991..3467
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3249..3268
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    3499..4087
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3605..3616
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4014..4034
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4112..5383,5384..5404,5405..5410))
                     /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      5407..5416
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cgaggacata tggagttaaa taatgaaggg atgcagaaag aagagtagtc acgtagtgac
       61 agagacaaca cgcgtgcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
      121 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcggcct
      181 cagtgagcga gcgagcgcgc agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg
      241 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      301 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      361 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      421 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      481 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      541 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      601 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      661 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
      721 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
      781 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
      841 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
      901 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
      961 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1021 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1081 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1141 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1201 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1261 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1321 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1381 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1441 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1501 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     1561 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1621 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     1681 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     1741 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     1801 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     1861 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     1921 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     1981 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     2041 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     2101 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2161 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2221 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2281 aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2341 tttttaacca atagaccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga
     2401 tagagttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     2461 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca
     2521 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     2581 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     2641 cgaaaggagc gggcgctaag gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     2701 cacccgccgc gcttaatgcg ccgctacagg gcgcgtacta tggttgcttt gacgtatgcg
     2761 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc cctgcaggca
     2821 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
     2881 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
     2941 taggggttcc tgcggccgct cggtccgcac gtggttacct acaaaatcag aaggacaggg
     3001 aagggagcag tggttcacgc ctgtaatccc agcaatttgg gaggccaagg tgggtagatc
     3061 acctgagatt aggagttgga gaccagcctg gccaatatgg tgaaaccccg tctctaccaa
     3121 aaaaacaaaa attagctgag cctggtcatg catgcctgga atcccaacaa ctcgggaggc
     3181 tgaggcagga gaatcgcttg aacccaggag gcggagattg cagtgagcca agattgtgcc
     3241 actgcactcc agcttggttc ccaatagacc ccgcaggccc tacaggttgt cttcccaact
     3301 tgccccttgc tccataccac ccccctccac cccataatat tatagaagga cacctagtca
     3361 gacaaaatga tgcaacttaa ttttattagg acaaggctgg tgggcactgg agtggcaact
     3421 tccagggcca ggagaggcac tggggagggg tcacagggat gccacccgta gatctctcga
     3481 gcagcgctcg gtatcgatgc ggggaggcgg cccaaaggga gatccgactc gtctgagggc
     3541 gaaggcgaag acgcggaaga ggccgcagag ccggcagcag gccgcgggaa ggaaggtccg
     3601 ctggattgag ggccgaaggg acgtagcaga aggacgtccc gcgcagaatc caggtggcaa
     3661 cataggcgag cagccaagga aaggacgatg atttccccga caacaccacg gaattgtcag
     3721 tgcccaacag ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg
     3781 tggcaatagg gagggggaaa gcgaaagtcc cggaaaggag ctgacaggtg gtggcaatgc
     3841 cccaaccagt gggggttgcg tcagcaaaca cagtgcacac cacgccacgt tgcctgacaa
     3901 cgggccacaa ctcctcataa agagacagca accaggattt atacaaggag gagaaaatga
     3961 aagccatacg ggaagcaata gcatgataca aaggcattaa agcagcgtat ccacatagcg
     4021 taaaaggagc aacatagtta agaataccag tcaatctttc acaaattttg taatccagag
     4081 gttgattatc gataagcttg atatcgaatt ctcagatccg cctgttgatg tagctgctca
     4141 ggtagtccag cacctcctgg ctgatgatgc cgttccaggc ggggtatctg atgctgccct
     4201 cggcgctgcc cttcagctgc tcgatgtgct gccactcctc gatggggttg gtctcgtcct
     4261 tcagggcgat catctccttg ctgatggggt cgtaggcgta gtacctggac accagggcga
     4321 agtagtggtc ggggatggcg gtgatctggt gggtgtaggt ggtcctggcc acggcggagg
     4381 ccctcttgtc gctccagttg cccaccacgt ttgtcagctc ggtcaggccc ttcatgctca
     4441 gaaagctggt catcaggtgc ctgccgatgt ggctcttagg gccgttcttg atagcgaaga
     4501 tggggtaggg ggcgttcttc ttcagggcct tgttgtagct gcgcaccagg ttgtccttca
     4561 gcagctggta ctcctgcttg ttgctgctgc tgttgccggt cctgttcact ctcttcagca
     4621 cgggctcgct gttcctcagg aactcgtcca ggtacaccag ggggtcgatc ctgcctctgg
     4681 cgctgaaaaa gtagatgtgc ctggacacgc ttgtcttggt ctcggtcacc aggcactgaa
     4741 tgatcacgcc caggtacttg ttctgcacca gcttgaagct cttggggtcc acgttcttga
     4801 tgtcgctgaa cctgccgcag ttgatgaatg tggccaggaa caggaactgg tacagggtct
     4861 tggtcttggt gaacctgctg gtgtactcga agctgttcag gatcttctcg gtgatctccc
     4921 agatgctctc gccctcggac agcagggcct tcagcatctt cttgctgtgg ctgttgccct
     4981 tgtcggcctc ctcgctgctc tcgaactgca gctgcaggct ggacacgatg tcggtgatgt
     5041 cgctctggtg cttctggccg ttgtaaggga tgatggtgaa ctcccaggcg gggatcagct
     5101 tcttcaggct ggcctccagg atggtggcct tctgggtctt gtacttgaac tgcaggctct
     5161 tgttcacgat gtcgaagctc aggctgttgc tgatgatggt gttgtagctc atgaaggtgg
     5221 ccctcttgat ggcggtgccg ttgtgggtga tcatccagca caggtaggtc agctcggcgg
     5281 cacagctggc gatcttctcg ccgctgggcc tctcgaatct ctccacgaac tgccgcacca
     5341 gcaccttggg gggggtcttg cacaggatgt cgaactggct catcaccttc ctcttcttct
     5401 taggagccat ggtggccacg ccagaagcaa atgtaagcaa tagatggctc tgccctgact
     5461 tttatgccca gcccgagctc cttaaggaag cattctggaa gtgcttgcct ttaatataac
     5521 aggctcgact cagacttgct tcagagttca tgctctctgg gtttgccgag gtaatgactt
     5581 ttagctcttt tttggtattg cttacattaa gactctgtag ctcatgtttt atggggattg
     5641 gtgcctgtaa cagatgcttt gtaaaatgcc aaggtagttt tctgccgcaa tcactgagtc
     5701 agagtttaaa ttcaacatct ggcatgattt gtcagcacct gaggtaaata acgctcagtg
     5761 attcagttgt ctggcccaat aaacatattc atgtgaaagt gtcattgaga gatgtcatga
     5821 tttattacaa gtcacacatc tccaaaacac gccatcacct aggtttcaat agaatattta
     5881 tgccgagaac aatactagca attcccacat catgtttctt gtaatctaag tactggagag
     5941 gcaga
//
