LOCUS       sequence-471215-        4672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2352m 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 4672)
  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 4672)
  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: AiP1763 - pAAV-AiE2352m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..126
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      684..693
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(690..692,693..695,696..1406)
                     /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(735..756)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(996..1015)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1343..1364
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1499..1519)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1706..1817
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1935..2075
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1935..2064
                     /label=AAV2 ITR
     rep_origin      2150..2605
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2237..2256)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2447..2468
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2622..2641)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2741..2763
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2801..2819)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2887..2991
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2992..3060,3061..3852)
                     /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     complement(3210..3229)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4023..4611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4512..4531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
      121 gttcctgcgg ccgcacgcgt tgtaagagta ataatcgagc ctccctggca cacagcacag
      181 tgactcaggt cagactacag ccgctcagaa caaacccttc aaaacaggac ctcaaaaatg
      241 tcaatgacta aggtactatt gttctcaaat gaccttggtt aggggcagag accactcgtg
      301 ttcataaaat aaaaagcaat gcaagatgag aacgttatgc tgattgactt cactgttttc
      361 aagatttaaa aataacattt tttgaaattg ttttgctttt gtttttgttt ttatagcgtt
      421 atcagctgat tatgggatag tctgagaagt ggaagtactc cttcaaatct tgtttcttcc
      481 tcttctgaag gacataaaag ctaagccttc agtgaagttg ccctctgaag gccaacggga
      541 agtatttgtt ttgtcttatg tgtaaaaccg gagttagcat ccaaaagacg gagctcgggc
      601 tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg gatccagatc
      661 tttcgaagct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
      721 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      781 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg atctgcacca
      841 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggcgtgcagt
      901 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
      961 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1021 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1081 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1141 tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca
     1201 acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1261 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag ctgagcaaag
     1321 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1381 ctctcggcat ggacgagctg tacaagtaag tcgacggcgc gccgcggccg cgaattcgat
     1441 atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     1501 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     1561 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg
     1621 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     1681 attccgtggc tcgagagatc ttcgactgtg ccttctagtt gccagccatc tgttgtttgc
     1741 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1801 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     1861 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat ctcacgtgcg
     1921 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     1981 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     2041 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     2101 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     2161 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2221 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2281 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2341 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     2401 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2461 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     2521 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2581 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     2641 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     2701 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     2761 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     2821 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     2881 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2941 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3001 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3061 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3121 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3181 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3241 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3301 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3361 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3421 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3481 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3541 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3601 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3661 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3721 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3781 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3841 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3901 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3961 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4021 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4081 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4141 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4201 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4261 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4321 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4381 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4441 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4501 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4561 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4621 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
