LOCUS       sequence-432586-        4620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2583m 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 4620)
  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 4620)
  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: AiP1895 - pAAV-AiE2583m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     127..146
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   308..448
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   319..448
                     /label=AAV2 ITR
     polyA_signal    566..677
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     864..884
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(977..1687,1688..1690,1691..1693))
                     /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(1019..1040)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1368..1387
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1627..1648
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1690..1699
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2201..2330
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2392..2980)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2472..2491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3151..3942,3943..4011))
                     /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     3774..3793
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4012..4116)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4184..4202
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4240..4262)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4362..4381
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4398..4620,1..233))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4535..4556)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
       61 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
      121 aaaggaaggg aagaaagcga aaggagcggg cgctaaggcg ctggcaagtg tagcggtcac
      181 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtactatgg
      241 ttgctttgac gtatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      301 aggcgcccct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      361 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      421 gtggccaact ccatcactag gggttcctgc ggccgctcgg tccgcacgtg agatctcatg
      481 cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat
      541 agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt
      601 gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg
      661 gcaactagaa ggcacagtcg aagatctctc gagccacgga attgtcagtg cccaacagcc
      721 gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg gcaagaacta
      781 accaggattt atacaaggag gagaaaatga aagccatacg ggaagcaata gcatgataca
      841 aaggcattaa agcagcgtat ccacatagcg taaaaggagc aacatagtta agaataccag
      901 tcaatctttc acaaattttg taatccagag gttgattatg atatcgaatt cgcggccgcg
      961 gcgcgccgtc gacttacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     1021 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agcttggact
     1081 ggtagctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     1141 gctggtagtg gtcggcgagc tgcacgccgc cgtcctcgat gttgtggcgg atcttgaagt
     1201 tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgtgg ctgttgtagt
     1261 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     1321 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     1381 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     1441 agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcactgcacg ccgtagccca
     1501 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atcagcttca
     1561 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     1621 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     1681 ccttgctcac catggtggcg accggtagcg ctagcttcga aagatctgga tcccagaagc
     1741 aaatgtaagc aatagatggc tctgccctga cttttatgcc cagcccgagc tccgaccaga
     1801 cagtgtcatc cgtgaagacc agcatcagca aagcccaatg ctgaccagca aagaatagca
     1861 aggcattcca ctaccattca gtatcatcta ctgtctgttg agctatattt atatgctttc
     1921 caaacattac ttatcctctc aagcgtctga tctagcaaaa caacacatgc cctttttcta
     1981 ggcagcttcc agaaaaatac cacatgtctg ttcttagcaa agcaccctcc cacatgtctg
     2041 cttcagcaaa aaacatcctc tcataagaca gttcccagaa acacatcaca ggacacaact
     2101 gagactccaa gaaaatcaca aatttccact tcatgagcat tttcaattgt atgagtactg
     2161 ccatgatttt atctttagag ttatattggt gtgcggccgc aggaacccct agtgatggag
     2221 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     2281 cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc
     2341 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2401 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2461 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2521 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2581 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2641 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2701 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2761 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2821 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2881 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2941 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3001 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3061 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3121 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3181 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3241 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3301 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3361 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3421 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3481 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3541 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3601 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3661 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3721 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3781 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3841 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3901 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3961 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4021 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4081 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4141 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     4201 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     4261 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     4321 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     4381 actgagagtg caccataaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt
     4441 ttgttaaatc agctcatttt ttaaccaata gaccgaaatc ggcaaaatcc cttataaatc
     4501 aaaagaatag cccgagatag agttgagtgt tgttccagtt tggaacaaga gtccactatt
     4561 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
//
