LOCUS       sequence-432554-        4565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2395m 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 4565)
  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 4565)
  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: AiP1726 - pAAV-AiE2395m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..46
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   208..348
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   219..348
                     /label=AAV2 ITR
     polyA_signal    466..577
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     764..784
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(877..1587,1588..1590,1591..1593))
                     /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(919..940)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1268..1287
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1527..1548
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1590..1599
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2046..2175
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2237..2825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2317..2336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2996..3787,3788..3856))
                     /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     3619..3638
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3857..3961)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4029..4047
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4085..4107)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4207..4226
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4243..4565,1..133))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4380..4401)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctaaggcg
       61 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
      121 ctacagggcg cgtactatgg ttgctttgac gtatgcggtg tgaaataccg cacagatgcg
      181 taaggagaaa ataccgcatc aggcgcccct gcaggcagct gcgcgctcgc tcgctcactg
      241 aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
      301 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc ggccgctcgg
      361 tccgcacgtg agatctcatg cctgctattg tcttcccaat cctccccctt gctgtcctgc
      421 cccaccccac cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca
      481 ttttattagg aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag
      541 gggcaaacaa cagatggctg gcaactagaa ggcacagtcg aagatctctc gagccacgga
      601 attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag
      661 ttccgccgtg gcaagaacta accaggattt atacaaggag gagaaaatga aagccatacg
      721 ggaagcaata gcatgataca aaggcattaa agcagcgtat ccacatagcg taaaaggagc
      781 aacatagtta agaataccag tcaatctttc acaaattttg taatccagag gttgattatg
      841 atatcgaatt cgcggccgcg gcgcgccgtc gacttacttg tacagctcgt ccatgccgag
      901 agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg
      961 gtctttgctc agcttggact ggtagctcag gtagtggttg tcgggcagca gcacggggcc
     1021 gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgccgc cgtcctcgat
     1081 gttgtggcgg atcttgaagt tggccttgat gccgttcttc tgcttgtcgg cggtgatata
     1141 gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt
     1201 gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc
     1261 ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc
     1321 ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa
     1381 gcactgcacg ccgtagccca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc
     1441 ggtggtgcag atcagcttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc
     1501 ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac
     1561 cccggtgaac agctcctcgc ccttgctcac catggtggcg accggtagcg ctagcttcga
     1621 aagatctgga tcccagaagc aaatgtaagc aatagatggc tctgccctga cttttatgcc
     1681 cagcccgagc tccgtgaatg caaagcagta tgtaataaat tatgtgtctt tttaagcaat
     1741 gaataaatgt taagcaaaca gatatgatga tactttttga ttgctctctc tctctctctc
     1801 tttctcaaaa aaaaatccca gctccctctc actgtgtttg cttgccagga agcttcagtg
     1861 gtgcctttgt gtaagcagtt tctggtgttg agatttaaaa ataaaacagg gcttactttc
     1921 tagcacgctt gtcctttcaa ggatagagct gtagctcaaa catccatcca gagtctggag
     1981 ggttcactaa ggccttttca tttctccgag cgcacgatta ttactcttac aacgcgtgcg
     2041 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     2101 tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc ctcagtgagc gagcgagcgc
     2161 gcagctgcct gcaggacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2221 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2281 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2341 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2401 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2461 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2521 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2581 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2641 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2701 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2761 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2821 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2881 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2941 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3001 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3061 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3121 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3181 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3241 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3301 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3361 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3421 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3481 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3541 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3601 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3661 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3721 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     3781 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     3841 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     3901 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     3961 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     4021 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     4081 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     4141 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     4201 actatgcggc atcagagcag attgtactga gagtgcacca taaaattgta aacgttaata
     4261 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caatagaccg
     4321 aaatcggcaa aatcccttat aaatcaaaag aatagcccga gatagagttg agtgttgttc
     4381 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4441 ccgtctatca gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt
     4501 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4561 gggga
//
