LOCUS       sequence-466356-        4684 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3087m 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 4684)
  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 4684)
  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: AiP20045 - pAAV-AiE3087m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   76..205
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(267..855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(347..366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1026..1817,1818..1886))
                     /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     1649..1668
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1887..1991)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2059..2077
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2115..2137)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2237..2256
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2273..2728)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2410..2431)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2622..2641
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2803..2943
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2814..2943
                     /label=AAV2 ITR
     polyA_signal    3061..3172
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3359..3379
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3472..4182,4183..4185,4186..4188))
                     /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(3514..3535)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3863..3882
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4122..4143
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4185..4194
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctttcatgat attaagatat ggagatttgg gcttctggga gggactgttt acaagtctaa
       61 aacgcgtgcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
      121 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc ctcagtgagc
      181 gagcgagcgc gcagctgcct gcaggacatg tgagcaaaag gccagcaaaa ggccaggaac
      241 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
      301 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
      361 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
      421 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
      481 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
      541 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
      601 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
      661 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
      721 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
      781 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
      841 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
      901 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
      961 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     1021 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     1081 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     1141 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     1201 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     1261 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     1321 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     1381 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     1441 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     1501 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     1561 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     1621 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     1681 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     1741 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     1801 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     1861 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     1921 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     1981 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     2041 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     2101 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     2161 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     2221 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca taaaattgta
     2281 aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     2341 caatagaccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga gatagagttg
     2401 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     2461 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc caaatcaagt
     2521 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     2581 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     2641 gcgggcgcta aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     2701 gcgcttaatg cgccgctaca gggcgcgtac tatggttgct ttgacgtatg cggtgtgaaa
     2761 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg cccctgcagg cagctgcgcg
     2821 ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc
     2881 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     2941 cctgcggccg ctcggtccgc acgtgagatc tcatgcctgc tattgtcttc ccaatcctcc
     3001 cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc
     3061 gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag
     3121 gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaagat
     3181 ctctcgagcc acggaattgt cagtgcccaa cagccgagcc cctgtccagc agcgggcaag
     3241 gcaggcggcg atgagttccg ccgtggcaag aactaaccag gatttataca aggaggagaa
     3301 aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag cgtatccaca
     3361 tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa ttttgtaatc
     3421 cagaggttga ttatgatatc gaattcgcgg ccgcggcgcg ccgtcgactt acttgtacag
     3481 ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc
     3541 gcgcttctcg ttggggtctt tgctcagctt ggactggtag ctcaggtagt ggttgtcggg
     3601 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac
     3661 gccgccgtcc tcgatgttgt ggcggatctt gaagttggcc ttgatgccgt tcttctgctt
     3721 gtcggcggtg atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat
     3781 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc
     3841 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg
     3901 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
     3961 ggggtagcgg gcgaagcact gcacgccgta gcccagggtg gtcacgaggg tgggccaggg
     4021 cacgggcagc ttgccggtgg tgcagatcag cttcagggtc agcttgccgt aggtggcatc
     4081 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac
     4141 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg
     4201 tagcgctagc ttcgaaagat ctggatccca gaagcaaatg taagcaatag atggctctgc
     4261 cctgactttt atgcccagcc cgagctccgc cggcgatttc tctctgatga aaccgatagt
     4321 atgcagggct ccttccgaag ctctttgttg ccgaagagca ggctgctcgg taggccagaa
     4381 cagattgtca ccatgtaaat gtggagtcaa gggacataat tttacgggct cacacctatc
     4441 tctcagccaa aggcatttca ggttagttgt ttgccgccac tggtgaagga atatggtatc
     4501 atttgaactg ggacaaccct agctccgggt acattctgtg gtatgatgtt tgccaactca
     4561 cattcagccc gtgcatttat aattgtcaac tgtttccatg cattgagtca tggcagaaaa
     4621 gcctcttctt tgtagaacct cattaagatt tcagttgcta acagatttta ttttacagtt
     4681 ccag
//
