LOCUS       sequence-432596-        4832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2543m 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 4832)
  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 4832)
  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: AiP1924 - pAAV-AiE2543m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4832
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   55..195
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   66..195
                     /label=AAV2 ITR
     polyA_signal    313..424
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     611..631
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(724..1434,1435..1437,1438..1440))
                     /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(766..787)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1115..1134
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1374..1395
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1437..1446
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2160..2289
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2351..2939)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2431..2450)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3110..3901,3902..3970))
                     /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     3733..3752
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3971..4075)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4143..4161
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4199..4221)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4321..4340
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4357..4812)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4494..4515)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4706..4725
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca
       61 ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga
      121 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      181 tcactagggg ttcctgcggc cgctcggtcc gcacgtgaga tctcatgcct gctattgtct
      241 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
      301 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
      361 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
      421 acagtcgaag atctctcgag ccacggaatt gtcagtgccc aacagccgag cccctgtcca
      481 gcagcgggca aggcaggcgg cgatgagttc cgccgtggca agaactaacc aggatttata
      541 caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc
      601 agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca
      661 aattttgtaa tccagaggtt gattatgata tcgaattcgc ggccgcggcg cgccgtcgac
      721 ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag
      781 gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggt agctcaggta
      841 gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc
      901 ggcgagctgc acgccgccgt cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc
      961 gttcttctgc ttgtcggcgg tgatatagac gttgtggctg ttgtagttgt actccagctt
     1021 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     1081 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     1141 gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     1201 cttcatgtgg tcggggtagc gggcgaagca ctgcacgccg tagcccaggg tggtcacgag
     1261 ggtgggccag ggcacgggca gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc
     1321 gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     1381 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
     1441 ggtggcgacc ggtagcgcta gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat
     1501 agatggctct gccctgactt ttatgcccag cccgagctcc gctagctgct ctgagaaatc
     1561 accaactatg tcctcatggg agttaagaaa agtacaaaaa agacaataat atgcaacaca
     1621 ttatataaac atcattgctt atattatgta atctctatgt cttttgttcc cagtatagaa
     1681 tgcactgctt gcataatttc actatataaa cagaacacta gaaagtagta agaggggctt
     1741 gctgtgatga actccttgat gggaagcagg aagaggaagg gtttgtctgg cttacaagga
     1801 actccaggtt acaatgtatc acttcagaga agtcaaggca ggacccccaa gcagctattc
     1861 atgtcacatg acaatcagga tagagaactg aattcactca ggccacctgc tgctactgct
     1921 gctgctgctg ctcagccagc actcttcact ctgactcagc tcagtgccca gcccacaggc
     1981 tagtgctgcc cacattcatg aggcatcttc ccacctcaac catcaagaca atcccccaca
     2041 gatgtcccaa cagccaccct gatgacgcta agttgtgata agttggtagt taaaactaga
     2101 cagcactcgt ggggagagga ataaaggatg acagagggac agagtaatga tgcggccgca
     2161 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     2221 cgggcgacca aaggtcgccc gacgcccggg cggcctcagt gagcgagcga gcgcgcagct
     2281 gcctgcagga catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2341 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2401 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2461 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2521 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2581 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2641 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2701 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2761 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2821 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2881 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2941 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3001 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3061 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3121 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3181 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3241 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3301 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3361 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3421 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3481 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3541 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3601 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3661 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3721 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3781 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3841 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3901 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3961 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4021 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4081 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     4141 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     4201 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     4261 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     4321 cggcatcaga gcagattgta ctgagagtgc accataaaat tgtaaacgtt aatattttgt
     4381 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag accgaaatcg
     4441 gcaaaatccc ttataaatca aaagaatagc ccgagataga gttgagtgtt gttccagttt
     4501 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4561 atcagggcga tggcccacta cgtgaaccat cacccaaatc aagttttttg gggtcgaggt
     4621 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4681 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctaaggcgc
     4741 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4801 tacagggcgc gtactatggt tgctttgacg ta
//
