LOCUS       sequence-438940-        5059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2474m 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 5059)
  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 5059)
  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: AiP1844 - pAAV-AiE2474m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(526..1317,1318..1386))
                     /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     1149..1168
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1387..1491)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1559..1577
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1615..1637)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1737..1756
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1773..2228)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1910..1931)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2122..2141
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2303..2443
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2314..2443
                     /label=AAV2 ITR
     polyA_signal    2561..2672
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2859..2879
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2972..3682,3683..3685,3686..3688))
                     /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(3014..3035)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3363..3382
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3622..3643
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3685..3694
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4635..4764
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4826..5059,1..355))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4906..4925)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
       61 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      121 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      181 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      241 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      301 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
      361 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
      421 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
      481 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
      541 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
      601 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
      661 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
      721 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
      781 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
      841 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
      901 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
      961 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1021 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1081 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1141 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1201 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1261 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1321 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     1381 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     1441 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     1501 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     1561 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     1621 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     1681 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     1741 atcagagcag attgtactga gagtgcacca taaaattgta aacgttaata ttttgttaaa
     1801 attcgcgtta aatttttgtt aaatcagctc attttttaac caatagaccg aaatcggcaa
     1861 aatcccttat aaatcaaaag aatagcccga gatagagttg agtgttgttc cagtttggaa
     1921 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     1981 gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg
     2041 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     2101 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta aggcgctggc
     2161 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     2221 gggcgcgtac tatggttgct ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg
     2281 agaaaatacc gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct cactgaggcc
     2341 gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
     2401 gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg ctcggtccgc
     2461 acgtgagatc tcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac
     2521 cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta
     2581 ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca
     2641 aacaacagat ggctggcaac tagaaggcac agtcgaagat ctctcgagcc acggaattgt
     2701 cagtgcccaa cagccgagcc cctgtccagc agcgggcaag gcaggcggcg atgagttccg
     2761 ccgtggcaag aactaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag
     2821 caatagcatg atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat
     2881 agttaagaat accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatgatatc
     2941 gaattcgcgg ccgcggcgcg ccgtcgactt acttgtacag ctcgtccatg ccgagagtga
     3001 tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt
     3061 tgctcagctt ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc
     3121 cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gccgccgtcc tcgatgttgt
     3181 ggcggatctt gaagttggcc ttgatgccgt tcttctgctt gtcggcggtg atatagacgt
     3241 tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt
     3301 cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc
     3361 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg
     3421 gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact
     3481 gcacgccgta gcccagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg
     3541 tgcagatcag cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca
     3601 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg
     3661 tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg tagcgctagc ttcgaaagat
     3721 ctggatccca gaagcaaatg taagcaatag atggctctgc cctgactttt atgcccagcc
     3781 cgagctccgg tattgtctgg tttttaggcc attttcaatt gacaaaaaga tcaatttgtt
     3841 ccagaatgtt tgattggaat atctgtctca aaactataac aaaattgtga tattgaaagg
     3901 aaattttgtt tatcttacat ctacaaatga tcggtcccat aagcagtggg ataatggtga
     3961 ctttggaggg aacatgcact tctcagtgca atgtgatgct ttggttatat ttattccaaa
     4021 gcctgatcat cattcatgtt cctcctttcc aggattcagc cattccattt taagccccta
     4081 ttcagccgca aatactccat tatgaagcag cctattggct aaaattggta tttatacagg
     4141 cgataagctt agaagcctat tttcttagca atgatcaaat ttagtctctc tgaatgatac
     4201 cctttgatgg ctttattctg cctactttca tatggattat ttctattatg cacaattaga
     4261 ctggcaagaa actccaaatg cagtttcaag actcataata aatgtactta ctaatgagaa
     4321 atgtggccat gctctgaata tttgtatttt tatgaacact taatcacaat cttatttgcc
     4381 atctctcaat atgacacatc atttgcctgc acaacaaacc atctaatagg ttaaaaaatc
     4441 attacctgat ataaaaagca gtggcccact ttcaatagtc tcttctcagt tccttaaatt
     4501 cgatgtctag ctaggaagag aatggtgctg agatggaatt taggaatgag ataaacgagg
     4561 caacctacag tccatggagt atcctcttgg cttcactacc atccgattat tactcttaca
     4621 acgcgtgcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     4681 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggcggcc tcagtgagcg
     4741 agcgagcgcg cagctgcctg caggacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4801 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4861 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4921 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4981 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5041 tcagttcggt gtaggtcgt
//
