LOCUS       sequence-432598-        4712 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2592m 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 4712)
  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 4712)
  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: AiP1963 - pAAV-AiE2592m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   115..255
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   126..255
                     /label=AAV2 ITR
     polyA_signal    373..484
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     671..691
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(784..1494,1495..1497,1498..1500))
                     /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(826..847)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1175..1194
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1434..1455
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1497..1506
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2100..2229
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2291..2879)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2371..2390)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3050..3841,3842..3910))
                     /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     3673..3692
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3911..4015)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4083..4101
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4139..4161)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4261..4280
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4297..4712,1..40))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4434..4455)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4646..4665
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgta
       61 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca
      121 ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga
      181 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      241 tcactagggg ttcctgcggc cgctcggtcc gcacgtgaga tctcatgcct gctattgtct
      301 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
      361 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
      421 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
      481 acagtcgaag atctctcgag ccacggaatt gtcagtgccc aacagccgag cccctgtcca
      541 gcagcgggca aggcaggcgg cgatgagttc cgccgtggca agaactaacc aggatttata
      601 caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc
      661 agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca
      721 aattttgtaa tccagaggtt gattatgata tcgaattcgc ggccgcggcg cgccgtcgac
      781 ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag
      841 gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggt agctcaggta
      901 gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc
      961 ggcgagctgc acgccgccgt cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc
     1021 gttcttctgc ttgtcggcgg tgatatagac gttgtggctg ttgtagttgt actccagctt
     1081 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     1141 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     1201 gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     1261 cttcatgtgg tcggggtagc gggcgaagca ctgcacgccg tagcccaggg tggtcacgag
     1321 ggtgggccag ggcacgggca gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc
     1381 gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     1441 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
     1501 ggtggcgacc ggtagcgcta gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat
     1561 agatggctct gccctgactt ttatgcccag cccgagctcc gcctcacagg ctgggtgtca
     1621 gcagatgtaa tcctggtgtg ttctctggga ccacctcaga cctgagtggg ctctcacaag
     1681 taagagaggg aaatcgggct cccaattttt cagaactcaa ttgcagttgg catcctggat
     1741 tctattctta cacaattgta agatcgggtg cagctcctgc ctttctgcga aaagcctctg
     1801 ggaaaaagat ttgtaatttc tttccacgac gacttcccag gggcctatca tctactgatg
     1861 cttttggtcg tgaagtggcg ccagagagtg tctcaggcac tcccaagctc tggctggctg
     1921 agccatgcgg gctgggctcg caccttccca agagctcaga tcaccctttc cagtctagtc
     1981 cctctaaagt tatgggattt cgctgggtga tgaggatctt ggctgagaaa aagactctta
     2041 ccaagcaaga aggagagagg gggctctctc tgctgtagtg cgcacaatcc tgcggccgca
     2101 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     2161 cgggcgacca aaggtcgccc gacgcccggg cggcctcagt gagcgagcga gcgcgcagct
     2221 gcctgcagga catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2281 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2341 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2401 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2461 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2521 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2581 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2641 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2701 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2761 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2821 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2881 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2941 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3001 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3061 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3121 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3181 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3241 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3301 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3361 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3421 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3481 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3541 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3601 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3661 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3721 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3781 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3841 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3901 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3961 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4021 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     4081 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     4141 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     4201 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     4261 cggcatcaga gcagattgta ctgagagtgc accataaaat tgtaaacgtt aatattttgt
     4321 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag accgaaatcg
     4381 gcaaaatccc ttataaatca aaagaatagc ccgagataga gttgagtgtt gttccagttt
     4441 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4501 atcagggcga tggcccacta cgtgaaccat cacccaaatc aagttttttg gggtcgaggt
     4561 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4621 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctaaggcgc
     4681 tggcaagtgt agcggtcacg ctgcgcgtaa cc
//
