LOCUS       sequence-432564-        4748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2459m 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 4748)
  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 4748)
  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: AiP1832 - pAAV-AiE2459m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(370..1161,1162..1230))
                     /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     993..1012
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1231..1335)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1403..1421
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1459..1481)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1581..1600
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1617..2072)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1754..1775)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1966..1985
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2147..2287
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2158..2287
                     /label=AAV2 ITR
     polyA_signal    2405..2516
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2703..2723
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2816..3526,3527..3529,3530..3532))
                     /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(2858..2879)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3207..3226
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3466..3487
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3529..3538
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4168..4297
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4359..4748,1..199))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4439..4458)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
       61 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
      121 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
      181 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
      241 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
      301 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
      361 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
      421 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
      481 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
      541 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
      601 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
      661 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
      721 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
      781 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
      841 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
      901 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
      961 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     1021 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     1081 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     1141 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     1201 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     1261 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     1321 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     1381 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     1441 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     1501 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     1561 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accataaaat
     1621 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     1681 taaccaatag accgaaatcg gcaaaatccc ttataaatca aaagaatagc ccgagataga
     1741 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     1801 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat cacccaaatc
     1861 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     1921 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     1981 aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     2041 cgccgcgctt aatgcgccgc tacagggcgc gtactatggt tgctttgacg tatgcggtgt
     2101 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgcccctg caggcagctg
     2161 cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt
     2221 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
     2281 ggttcctgcg gccgctcggt ccgcacgtga gatctcatgc ctgctattgt cttcccaatc
     2341 ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca
     2401 atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt
     2461 caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga
     2521 agatctctcg agccacggaa ttgtcagtgc ccaacagccg agcccctgtc cagcagcggg
     2581 caaggcaggc ggcgatgagt tccgccgtgg caagaactaa ccaggattta tacaaggagg
     2641 agaaaatgaa agccatacgg gaagcaatag catgatacaa aggcattaaa gcagcgtatc
     2701 cacatagcgt aaaaggagca acatagttaa gaataccagt caatctttca caaattttgt
     2761 aatccagagg ttgattatga tatcgaattc gcggccgcgg cgcgccgtcg acttacttgt
     2821 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
     2881 gatcgcgctt ctcgttgggg tctttgctca gcttggactg gtagctcagg tagtggttgt
     2941 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct
     3001 gcacgccgcc gtcctcgatg ttgtggcgga tcttgaagtt ggccttgatg ccgttcttct
     3061 gcttgtcggc ggtgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
     3121 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
     3181 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
     3241 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
     3301 ggtcggggta gcgggcgaag cactgcacgc cgtagcccag ggtggtcacg agggtgggcc
     3361 agggcacggg cagcttgccg gtggtgcaga tcagcttcag ggtcagcttg ccgtaggtgg
     3421 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     3481 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga
     3541 ccggtagcgc tagcttcgaa agatctggat cccagaagca aatgtaagca atagatggct
     3601 ctgccctgac ttttatgccc agcccgagct ccgttccttt tacatacata ttaattagtg
     3661 ttttcgaaaa tctcaacatc ttatccataa tattggcaaa gtagcttcac atcccagcta
     3721 caacagtata agaggcagag aaagaatggt tacttctggg aagcaagaag cctgtctgag
     3781 taatagtgtt tggcacactc tgaaatcaaa agcaatgaat ttgaatctta accagaaaga
     3841 ctaaaatgcc atcttatttt ggaaattact cagagctatt ttaaagcagc agcagagcca
     3901 aagcggcttc cctacaataa aatccactca acaaatagac tgttggagaa agattacaac
     3961 tctccaaaca tgtgtgagtt tctgtgacat gtgaatctat atttcacagc tacttttaag
     4021 gtaagtccat tttcatgagc ttgagtagaa caatggggaa aacaagcaag catacacaaa
     4081 gggatgtgtg caggttttac ttctgactga aaatcatttt taagaagaaa aaaaaacgat
     4141 tattactctt acaacgcgtg cggccgcagg aacccctagt gatggagttg gccactccct
     4201 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg
     4261 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa
     4321 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4381 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4441 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4501 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4561 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4621 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4681 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4741 tatgtagg
//
