LOCUS       sequence-433108-        4757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2536m 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 4757)
  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 4757)
  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: AiP1854 - pAAV-AiE2536m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     59..78
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   240..380
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   251..380
                     /label=AAV2 ITR
     polyA_signal    498..609
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     796..816
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(909..1619,1620..1622,1623..1625))
                     /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(951..972)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1300..1319
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1559..1580
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1622..1631
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2270..2399
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2461..3049)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2541..2560)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3220..4011,4012..4080))
                     /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     3843..3862
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4081..4185)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4253..4271
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4309..4331)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4431..4450
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4467..4757,1..165))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4604..4625)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
       61 ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag tgtagcggtc acgctgcgcg
      121 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtactat ggttgctttg
      181 acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgccc
      241 ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg
      301 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
      361 ctccatcact aggggttcct gcggccgctc ggtccgcacg tgagatctca tgcctgctat
      421 tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga atagaatgac
      481 acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg
      541 caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag
      601 aaggcacagt cgaagatctc tcgagccacg gaattgtcag tgcccaacag ccgagcccct
      661 gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaagaac taaccaggat
      721 ttatacaagg aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt
      781 aaagcagcgt atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt
      841 tcacaaattt tgtaatccag aggttgatta tgatatcgaa ttcgcggccg cggcgcgccg
      901 tcgacttact tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
      961 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga ctggtagctc
     1021 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
     1081 tggtcggcga gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa gttggccttg
     1141 atgccgttct tctgcttgtc ggcggtgata tagacgttgt ggctgttgta gttgtactcc
     1201 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg
     1261 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc
     1321 ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg
     1381 tgctgcttca tgtggtcggg gtagcgggcg aagcactgca cgccgtagcc cagggtggtc
     1441 acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatcagctt cagggtcagc
     1501 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg
     1561 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc
     1621 accatggtgg cgaccggtag cgctagcttc gaaagatctg gatcccagaa gcaaatgtaa
     1681 gcaatagatg gctctgccct gacttttatg cccagcccga gctccgttat aggctgtatg
     1741 accttaggaa agttactagt tctctctact attcaaatct catgaattgg aaaaagtagt
     1801 cataataata gaaagtgcct ggagccccta attaagtgat tcagacacat agtaattcgg
     1861 tgagtatttt tgtgtatcag ccgtatcact gaatactttg gtctctccaa tctaataata
     1921 gggttgggtt cagagtcata gaataacaac tagagcagaa tatccagaga ccttcaagac
     1981 cagctccact ggcctccggg gtggggggtg ggggtaatga cttgcccagc gtcacagtta
     2041 atccaaagtc ctgactgctc ccctttgccc ctctctctgc ccagtcctct cacacgcggc
     2101 atcaaggtca ctgtgcatca gatctgggaa tgcacgcttt gtgcctttaa gatgtcccaa
     2161 ggttctacgc gctcagttaa acccagccag agagggttct gctctaacaa acagcactcg
     2221 acagccatgg ttgatttgcg attattactc ttacaacgcg tgcggccgca ggaaccccta
     2281 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     2341 aaggtcgccc gacgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcagga
     2401 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2461 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2521 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2581 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2641 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2701 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2761 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2821 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2881 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2941 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3001 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3061 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3121 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3181 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3241 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3301 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3361 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3421 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3481 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3541 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3601 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3661 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3721 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3781 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3841 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3901 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3961 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4021 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4081 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4141 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4201 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     4261 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     4321 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     4381 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     4441 gcagattgta ctgagagtgc accataaaat tgtaaacgtt aatattttgt taaaattcgc
     4501 gttaaatttt tgttaaatca gctcattttt taaccaatag accgaaatcg gcaaaatccc
     4561 ttataaatca aaagaatagc ccgagataga gttgagtgtt gttccagttt ggaacaagag
     4621 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4681 tggcccacta cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc
     4741 actaaatcgg aacccta
//
