LOCUS       sequence-466281-        4746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2332m 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 4746)
  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 4746)
  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: AiP1652 - pAAV-AiE2332m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      479..1067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     968..987
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1129..1258
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1886..1895
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1892..1894,1895..1897,1898..2608)
                     /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(1937..1958)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2198..2217)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2545..2566
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2701..2721)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2908..3019
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3137..3277
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3137..3266
                     /label=AAV2 ITR
     rep_origin      3352..3807
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3439..3458)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3649..3670
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3824..3843)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3943..3965
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4003..4021)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4089..4193
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4194..4262,4263..4746,1..308)
                     /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     complement(4412..4431)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
       61 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
      121 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
      181 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
      241 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
      301 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
      361 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
      421 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
      481 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
      541 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
      601 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
      661 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
      721 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
      781 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
      841 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
      901 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      961 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     1021 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     1081 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc
     1141 tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc
     1201 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
     1261 ggccgcacgc gttgtaagag taataatcgg ttctggagac attgcccagc tttccagcag
     1321 cctgccagac tctgaaccca gagtgcagac tcattagcat tcagaggggg actaaccatg
     1381 ggggacacat ctcaagcact aggaagagac aggctgtgtt ttaattatga cctggacttt
     1441 tgaacaaatc cttttctctg cttctctgag cctcagagaa aacaacttat gggaggctgt
     1501 gcagtcggca gctgggtcca cagtggtacc cgtacagctg tgtacacatc tggcacccag
     1561 gtggtagatg gcatgttcct cttctcgaat tgctcttgaa ggttacaaga gacaagggag
     1621 taaggtgcag ctgacgtggt ttggctccag ggagggagaa cggaaggaac aaggcttgct
     1681 ggactctctg gcctgcagtc ctccagggaa catggccaat tcaggatttg agcctagatg
     1741 actacttaca cttagtctag cgctcctgag tcaacatggc ggacactctt cggagctcgg
     1801 gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga
     1861 tctttcgaag ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac
     1921 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1981 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac
     2041 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca
     2101 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2161 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2221 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2281 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     2341 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
     2401 caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2461 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa
     2521 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2581 cactctcggc atggacgagc tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg
     2641 atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2701 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2761 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc
     2821 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2881 caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt
     2941 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3001 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3061 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg
     3121 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3181 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3241 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     3301 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     3361 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3421 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3481 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3541 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     3601 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3661 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     3721 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3781 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     3841 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3901 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3961 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     4021 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4081 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4141 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4201 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4261 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4321 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4381 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4441 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4501 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4561 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4621 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4681 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4741 caatgg
//
