LOCUS       sequence-432582-        4520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2522m 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 4520)
  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 4520)
  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: AiP1939 - pAAV-AiE2522m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(12..116)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     184..202
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(240..262)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     362..381
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(398..853)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(535..556)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     747..766
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   928..1068
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   939..1068
                     /label=AAV2 ITR
     polyA_signal    1186..1297
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1484..1504
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1597..2307,2308..2310,2311..2313))
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1639..1660)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1988..2007
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2247..2268
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2310..2319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2721..2850
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2912..3500)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2992..3011)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3671..4462,4463..4520,1..11))
                     /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     4294..4313
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
       61 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
      121 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
      181 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
      241 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
      301 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
      361 gcggcatcag agcagattgt actgagagtg caccataaaa ttgtaaacgt taatattttg
      421 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata gaccgaaatc
      481 ggcaaaatcc cttataaatc aaaagaatag cccgagatag agttgagtgt tgttccagtt
      541 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
      601 tatcagggcg atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg
      661 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
      721 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctaaggcg
      781 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
      841 ctacagggcg cgtactatgg ttgctttgac gtatgcggtg tgaaataccg cacagatgcg
      901 taaggagaaa ataccgcatc aggcgcccct gcaggcagct gcgcgctcgc tcgctcactg
      961 aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
     1021 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc ggccgctcgg
     1081 tccgcacgtg agatctcatg cctgctattg tcttcccaat cctccccctt gctgtcctgc
     1141 cccaccccac cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca
     1201 ttttattagg aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag
     1261 gggcaaacaa cagatggctg gcaactagaa ggcacagtcg aagatctctc gagccacgga
     1321 attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag
     1381 ttccgccgtg gcaagaacta accaggattt atacaaggag gagaaaatga aagccatacg
     1441 ggaagcaata gcatgataca aaggcattaa agcagcgtat ccacatagcg taaaaggagc
     1501 aacatagtta agaataccag tcaatctttc acaaattttg taatccagag gttgattatg
     1561 atatcgaatt cgcggccgcg gcgcgccgtc gacttacttg tacagctcgt ccatgccgag
     1621 agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg
     1681 gtctttgctc agcttggact ggtagctcag gtagtggttg tcgggcagca gcacggggcc
     1741 gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgccgc cgtcctcgat
     1801 gttgtggcgg atcttgaagt tggccttgat gccgttcttc tgcttgtcgg cggtgatata
     1861 gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt
     1921 gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc
     1981 ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc
     2041 ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa
     2101 gcactgcacg ccgtagccca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc
     2161 ggtggtgcag atcagcttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc
     2221 ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac
     2281 cccggtgaac agctcctcgc ccttgctcac catggtggcg accggtagcg ctagcttcga
     2341 aagatctgga tcccagaagc aaatgtaagc aatagatggc tctgccctga cttttatgcc
     2401 cagcccgagc tccgagaaca gaactaatcc tgagtaattg taacagcagc aagggaccag
     2461 gacaacgcat gcttttcttt gctatggaca gattgagtga gcaaatgaat gggtgacacc
     2521 ccctccattc agaccctttc agatcccagc agaggttggg tctgagtcag ccacacagga
     2581 gagggagggg agggtgctct gtacaagcca ttctcgaaga acgttaatca cacgccaagc
     2641 cagattctga gcaggacttg gcactttcag cttctaaaaa tgtaggaaac gattattact
     2701 cttacaacgc gtgcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     2761 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag
     2821 tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc aaaaggccag caaaaggcca
     2881 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2941 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3001 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3061 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3121 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3181 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3241 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3301 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3361 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3421 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3481 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3541 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3601 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3661 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3721 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3781 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3841 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3901 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3961 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4021 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4081 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4141 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4201 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4261 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4321 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4381 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4441 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4501 taagggcgac acggaaatgt
//
