LOCUS       sequence-466207-        5886 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2046m 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 5886)
  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 5886)
  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: AiP1187 - pAAV-AiE2046m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..5886
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   331..460
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(522..1110)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(602..621)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1281..2072,2073..2141))
                     /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     1904..1923
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2142..2246)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2314..2332
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2370..2392)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2492..2511
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2528..2983)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2665..2686)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2877..2896
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3058..3198
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3069..3198
                     /label=AAV2 ITR
     polyA_signal    3238..3714
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3496..3515
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    3746..4334
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3852..3863
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4261..4281
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4359..5630,5631..5651,5652..5657))
                     /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      5654..5663
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cgtgaggaga atgtgacatc aagaacaact cctagggttc tgactcagag tacagataga
       61 gacgtccttt tgtagataga aaaggttgag tttcacaagt gagcagcaag ccacgagatc
      121 atgaggagat tgcacctgga gtattcagag gaagctttaa aaatgcaaat gaaaatttgg
      181 agatccgaat gaaggttact cagctcattt gaattgacca ggtatatcag tgttttcttg
      241 ttgctataac aagatagcct agactgggtg gtttaagcaa cagaaatgta ttttatttca
      301 cagttctgag ggacagacgc gtgcggccgc aggaacccct agtgatggag ttggccactc
      361 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
      421 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc aaaaggccag
      481 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      541 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      601 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      661 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      721 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      781 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      841 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      901 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      961 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1021 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1081 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1141 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1201 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     1261 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1321 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1381 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1441 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1501 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1561 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1621 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1681 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1741 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1801 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1861 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1921 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1981 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2041 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2101 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2161 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2221 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     2281 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     2341 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     2401 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     2461 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     2521 caccataaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     2581 agctcatttt ttaaccaata gaccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     2641 cccgagatag agttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     2701 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     2761 tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     2821 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     2881 aagaaagcga aaggagcggg cgctaaggcg ctggcaagtg tagcggtcac gctgcgcgta
     2941 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtactatgg ttgctttgac
     3001 gtatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgcccct
     3061 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg
     3121 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
     3181 ccatcactag gggttcctgc ggccgctcgg tccgcacgtg gttacctaca aaatcagaag
     3241 gacagggaag ggagcagtgg ttcacgcctg taatcccagc aatttgggag gccaaggtgg
     3301 gtagatcacc tgagattagg agttggagac cagcctggcc aatatggtga aaccccgtct
     3361 ctaccaaaaa aacaaaaatt agctgagcct ggtcatgcat gcctggaatc ccaacaactc
     3421 gggaggctga ggcaggagaa tcgcttgaac ccaggaggcg gagattgcag tgagccaaga
     3481 ttgtgccact gcactccagc ttggttccca atagaccccg caggccctac aggttgtctt
     3541 cccaacttgc cccttgctcc ataccacccc cctccacccc ataatattat agaaggacac
     3601 ctagtcagac aaaatgatgc aacttaattt tattaggaca aggctggtgg gcactggagt
     3661 ggcaacttcc agggccagga gaggcactgg ggaggggtca cagggatgcc acccgtagat
     3721 ctctcgagca gcgctcggta tcgatgcggg gaggcggccc aaagggagat ccgactcgtc
     3781 tgagggcgaa ggcgaagacg cggaagaggc cgcagagccg gcagcaggcc gcgggaagga
     3841 aggtccgctg gattgagggc cgaagggacg tagcagaagg acgtcccgcg cagaatccag
     3901 gtggcaacat aggcgagcag ccaaggaaag gacgatgatt tccccgacaa caccacggaa
     3961 ttgtcagtgc ccaacagccg agcccctgtc cagcagcggg caaggcaggc ggcgatgagt
     4021 tccgccgtgg caatagggag ggggaaagcg aaagtcccgg aaaggagctg acaggtggtg
     4081 gcaatgcccc aaccagtggg ggttgcgtca gcaaacacag tgcacaccac gccacgttgc
     4141 ctgacaacgg gccacaactc ctcataaaga gacagcaacc aggatttata caaggaggag
     4201 aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc agcgtatcca
     4261 catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca aattttgtaa
     4321 tccagaggtt gattatcgat aagcttgata tcgaattctc agatccgcct gttgatgtag
     4381 ctgctcaggt agtccagcac ctcctggctg atgatgccgt tccaggcggg gtatctgatg
     4441 ctgccctcgg cgctgccctt cagctgctcg atgtgctgcc actcctcgat ggggttggtc
     4501 tcgtccttca gggcgatcat ctccttgctg atggggtcgt aggcgtagta cctggacacc
     4561 agggcgaagt agtggtcggg gatggcggtg atctggtggg tgtaggtggt cctggccacg
     4621 gcggaggccc tcttgtcgct ccagttgccc accacgtttg tcagctcggt caggcccttc
     4681 atgctcagaa agctggtcat caggtgcctg ccgatgtggc tcttagggcc gttcttgata
     4741 gcgaagatgg ggtagggggc gttcttcttc agggccttgt tgtagctgcg caccaggttg
     4801 tccttcagca gctggtactc ctgcttgttg ctgctgctgt tgccggtcct gttcactctc
     4861 ttcagcacgg gctcgctgtt cctcaggaac tcgtccaggt acaccagggg gtcgatcctg
     4921 cctctggcgc tgaaaaagta gatgtgcctg gacacgcttg tcttggtctc ggtcaccagg
     4981 cactgaatga tcacgcccag gtacttgttc tgcaccagct tgaagctctt ggggtccacg
     5041 ttcttgatgt cgctgaacct gccgcagttg atgaatgtgg ccaggaacag gaactggtac
     5101 agggtcttgg tcttggtgaa cctgctggtg tactcgaagc tgttcaggat cttctcggtg
     5161 atctcccaga tgctctcgcc ctcggacagc agggccttca gcatcttctt gctgtggctg
     5221 ttgcccttgt cggcctcctc gctgctctcg aactgcagct gcaggctgga cacgatgtcg
     5281 gtgatgtcgc tctggtgctt ctggccgttg taagggatga tggtgaactc ccaggcgggg
     5341 atcagcttct tcaggctggc ctccaggatg gtggccttct gggtcttgta cttgaactgc
     5401 aggctcttgt tcacgatgtc gaagctcagg ctgttgctga tgatggtgtt gtagctcatg
     5461 aaggtggccc tcttgatggc ggtgccgttg tgggtgatca tccagcacag gtaggtcagc
     5521 tcggcggcac agctggcgat cttctcgccg ctgggcctct cgaatctctc cacgaactgc
     5581 cgcaccagca ccttgggggg ggtcttgcac aggatgtcga actggctcat caccttcctc
     5641 ttcttcttag gagccatggt ggccacgcca gaagcaaatg taagcaatag atggctctgc
     5701 cctgactttt atgcccagcc cgagctcctt aagctaattt cctctgcagg ctgcatccct
     5761 agccatgatt cccaaaactg aagtgatgtc tgagcatggt gcattaatga tagaaatgtg
     5821 ttcctctgca gcactgaaaa atatgaaaga caatgaaaca atgggatgag gagggtttaa
     5881 agtggg
//
