LOCUS       sequence-466307-        4808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2528m 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 4808)
  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 4808)
  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: AiP1785 - pAAV-AiE2528m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     79..98
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   260..400
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   271..400
                     /label=AAV2 ITR
     polyA_signal    518..629
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     816..836
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(929..1639,1640..1642,1643..1645))
                     /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(971..992)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1320..1339
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1579..1600
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1642..1651
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2341..2470
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2532..3120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2612..2631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3291..4082,4083..4151))
                     /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     3914..3933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4152..4256)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4324..4342
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4380..4402)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4502..4521
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4538..4808,1..185))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4675..4696)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
       61 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag
      121 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
      181 cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga
      241 aaataccgca tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
      301 cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
      361 cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgctc ggtccgcacg
      421 tgagatctca tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc
      481 accccccaga atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta
      541 ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac
      601 aacagatggc tggcaactag aaggcacagt cgaagatctc tcgagccacg gaattgtcag
      661 tgcccaacag ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg
      721 tggcaagaac taaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa
      781 tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt
      841 taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgatta tgatatcgaa
      901 ttcgcggccg cggcgcgccg tcgacttact tgtacagctc gtccatgccg agagtgatcc
      961 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
     1021 tcagcttgga ctggtagctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     1081 tgggggtgtt ctgctggtag tggtcggcga gctgcacgcc gccgtcctcg atgttgtggc
     1141 ggatcttgaa gttggccttg atgccgttct tctgcttgtc ggcggtgata tagacgttgt
     1201 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
     1261 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
     1321 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
     1381 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg aagcactgca
     1441 cgccgtagcc cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
     1501 agatcagctt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc
     1561 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
     1621 acagctcctc gcccttgctc accatggtgg cgaccggtag cgctagcttc gaaagatctg
     1681 gatcccagaa gcaaatgtaa gcaatagatg gctctgccct gacttttatg cccagcccga
     1741 gctccgagtt caacatggtg ggcaggcgtt gcagcagaca caggaagctg ggaattcaca
     1801 tttctatcca cacataggaa gcagagaaag tgactggagt taggtgaggc taccttttag
     1861 cctgccttca gtaacatact tcctccgtga aggctataca aaggctctat aacctcggca
     1921 aaccaacaac aagaagccag atatcggcat tcctgagcct aaaataagta gctctcattt
     1981 aaatgagcac accagcgtga aaacacatca tgccaagcaa tacatttgtt gaacaattat
     2041 ccaaagctgt gtatttttat attcactatt tatacatcta atatcatacc tctgaaaaac
     2101 aaaagacagc tggccaatgt gtgataagtg cttatagaaa ttgtctgtga ctgggagaga
     2161 aataactgag cagaacacat gcttagtatg aaaatcgaca tgctcatttt cagaggtttg
     2221 gaagaagtct ctggctcctt tgcacatgag agatgagcat cccagctgcg tggcacagag
     2281 ttcaaggata agcggtctgg aacaagaccc gattattact cttacaacgc gtgcggccgc
     2341 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     2401 ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     2461 tgcctgcagg acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2521 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2581 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2641 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2701 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2761 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2821 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2881 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2941 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3001 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3061 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3121 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3181 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3241 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3301 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3361 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3421 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3481 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3541 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3601 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3661 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3721 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3781 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3841 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3901 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     3961 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4021 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4081 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4141 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4201 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4261 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     4321 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     4381 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     4441 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     4501 gcggcatcag agcagattgt actgagagtg caccataaaa ttgtaaacgt taatattttg
     4561 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata gaccgaaatc
     4621 ggcaaaatcc cttataaatc aaaagaatag cccgagatag agttgagtgt tgttccagtt
     4681 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     4741 tatcagggcg atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg
     4801 tgccgtaa
//
