LOCUS       sequence-432529-        5001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2088m 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 5001)
  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 5001)
  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: AiP1272 - pAAV-AiE2088m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(101..205)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     273..291
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(329..351)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     451..470
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(487..942)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(624..645)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     836..855
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1017..1157
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1028..1157
                     /label=AAV2 ITR
     polyA_signal    1275..1386
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1573..1593
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1686..2396,2397..2399,2400..2402))
                     /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(1728..1749)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2077..2096
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2336..2357
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2399..2408
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3291..3420
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3482..4070)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3562..3581)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4241..5001,1..31,32..100))
                     /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     4864..4883
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
       61 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
      121 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
      181 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
      241 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
      301 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
      361 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      421 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      481 accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
      541 gctcattttt taaccaatag accgaaatcg gcaaaatccc ttataaatca aaagaatagc
      601 ccgagataga gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      661 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      721 cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      781 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      841 agaaagcgaa aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
      901 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt tgctttgacg
      961 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgcccctg
     1021 caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc
     1081 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
     1141 catcactagg ggttcctgcg gccgctcggt ccgcacgtga gatctcatgc ctgctattgt
     1201 cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc
     1261 tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac
     1321 cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag
     1381 gcacagtcga agatctctcg agccacggaa ttgtcagtgc ccaacagccg agcccctgtc
     1441 cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caagaactaa ccaggattta
     1501 tacaaggagg agaaaatgaa agccatacgg gaagcaatag catgatacaa aggcattaaa
     1561 gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt caatctttca
     1621 caaattttgt aatccagagg ttgattatga tatcgaattc gcggccgcgg cgcgccgtcg
     1681 acttacttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc
     1741 aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gcttggactg gtagctcagg
     1801 tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg
     1861 tcggcgagct gcacgccgcc gtcctcgatg ttgtggcgga tcttgaagtt ggccttgatg
     1921 ccgttcttct gcttgtcggc ggtgatatag acgttgtggc tgttgtagtt gtactccagc
     1981 ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc
     2041 accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg
     2101 aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc
     2161 tgcttcatgt ggtcggggta gcgggcgaag cactgcacgc cgtagcccag ggtggtcacg
     2221 agggtgggcc agggcacggg cagcttgccg gtggtgcaga tcagcttcag ggtcagcttg
     2281 ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg
     2341 ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc
     2401 atggtggcga ccggtagcgc tagcttcgaa agatctggat cccagaagca aatgtaagca
     2461 atagatggct ctgccctgac ttttatgccc agcccgagct ctgcttaaac agactatgac
     2521 ataaatgcaa aaaatgctgc atttatttta aaatttattc ttgacaactt cacaggccta
     2581 ttccctctta ttatatttct tctattcact tttaagtgta aatgatctta tctcaaatca
     2641 cacctttcaa ctaacaattg cagggaaaag cagtaataat ttaccttggt ttggaattat
     2701 tcttcaattt tccattcaca tttcactttt ctacaacata aaaatggacc aattgctaat
     2761 cagctgttaa acatgattag aatacttcta gacaacaaaa acgtgaattt cagactttta
     2821 gggtaattaa aagaactatt tgggcctcta attattgttg tttttatagc ctgataccag
     2881 ctgtccatgg acaaatgata aagtaggaaa ttcaagtaat tactcagata ttttagggaa
     2941 aatcaagtgt ttcagctaca catgttccac tcaacacttt gtttaaatta tttttatttt
     3001 tcccattttc atggagataa ggatattcaa gttccgaaca cattctgcat gttcttattt
     3061 agatgcatat aaacaaatct cttcattttt attgcataac ttacaatcaa atatttatat
     3121 taatctatag gacccagtag cctagaatat gaatacttaa aattacttag aaatttagac
     3181 ccaactaaga atagacagta aatatgatgt ggtagatagg agtataagat gcacagtctc
     3241 tttagaagat agtatcccac tttcctacta aaatgaacgc gtgcggccgc aggaacccct
     3301 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     3361 aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     3421 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3481 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3541 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3601 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3661 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3721 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3781 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3841 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3901 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3961 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4021 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4081 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4141 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4201 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4261 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4321 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4381 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4441 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4501 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4561 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4621 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4681 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4741 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4801 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4861 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4921 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4981 gtgcacccaa ctgatcttca g
//
