LOCUS       sequence-466353-        4677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3088m 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 4677)
  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 4677)
  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: AiP20041 - pAAV-AiE3088m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(348..1139,1140..1208))
                     /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     971..990
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1209..1313)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1381..1399
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1437..1459)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1559..1578
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1595..2050)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1732..1753)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1944..1963
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2125..2265
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2136..2265
                     /label=AAV2 ITR
     polyA_signal    2383..2494
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2681..2701
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2794..3504,3505..3507,3508..3510))
                     /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(2836..2857)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3185..3204
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3444..3465
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3507..3516
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4075..4204
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4266..4677,1..177))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4346..4365)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
       61 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      121 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      181 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
      241 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
      301 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
      361 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
      421 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
      481 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
      541 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
      601 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
      661 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
      721 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
      781 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
      841 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
      901 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
      961 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1021 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1081 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1141 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1201 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1261 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     1321 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     1381 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     1441 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     1501 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     1561 gcatcagagc agattgtact gagagtgcac cataaaattg taaacgttaa tattttgtta
     1621 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaatagac cgaaatcggc
     1681 aaaatccctt ataaatcaaa agaatagccc gagatagagt tgagtgttgt tccagtttgg
     1741 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     1801 cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc
     1861 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     1921 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc taaggcgctg
     1981 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2041 cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac agatgcgtaa
     2101 ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg cgctcgctcg ctcactgagg
     2161 ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc
     2221 gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc cgctcggtcc
     2281 gcacgtgaga tctcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc
     2341 accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt
     2401 tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg
     2461 caaacaacag atggctggca actagaaggc acagtcgaag atctctcgag ccacggaatt
     2521 gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc
     2581 cgccgtggca agaactaacc aggatttata caaggaggag aaaatgaaag ccatacggga
     2641 agcaatagca tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac
     2701 atagttaaga ataccagtca atctttcaca aattttgtaa tccagaggtt gattatgata
     2761 tcgaattcgc ggccgcggcg cgccgtcgac ttacttgtac agctcgtcca tgccgagagt
     2821 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
     2881 tttgctcagc ttggactggt agctcaggta gtggttgtcg ggcagcagca cggggccgtc
     2941 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt cctcgatgtt
     3001 gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg tgatatagac
     3061 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
     3121 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
     3181 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc
     3241 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc gggcgaagca
     3301 ctgcacgccg tagcccaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     3361 ggtgcagatc agcttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga
     3421 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     3481 ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtagcgcta gcttcgaaag
     3541 atctggatcc cagaagcaaa tgtaagcaat agatggctct gccctgactt ttatgcccag
     3601 cccgagctcc gccggcgcag aaattcagcc aaacttttat tattttttta aaatcgatgt
     3661 aaatctccca tgctgtttaa atgtcggaac taaatttacc aaacttggca gattcgtaaa
     3721 gctaagcagg actgccaaga gttcccagtt tcaagcagag agagttctaa cagctcctga
     3781 aatacttgct ggaggcacct ggtggttaat tgcagctcca tatttcctcc tgagcgccta
     3841 ccttgctcgc tggcattggc gcacatagga ttattacagc gatgatgagg tggcagaata
     3901 gaagtcctaa aataaggcac agccccggaa agggaacaca ttcatacaga ctcctcccta
     3961 aggaattagg catgttcctg atattctctc aagctggtta tgccaaacaa gtagaggggc
     4021 agttggcaag ggagacggaa accatacccg taaatccgta acgcgtgcgg ccgcaggaac
     4081 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     4141 gaccaaaggt cgcccgacgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     4201 caggacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4261 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4321 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4381 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4441 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4501 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4561 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4621 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaag
//
