LOCUS       sequence-461347-        4712 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0476m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific brain cell types.
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: AiP12321 - pAAV-AiE0476m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      724..733
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(730..732,733..735,736..1446)
                     /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(775..796)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1036..1055)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1383..1404
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1539..1559)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1746..1857
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1975..2115
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1975..2104
                     /label=AAV2 ITR
     rep_origin      2190..2645
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2277..2296)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2487..2508
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2662..2681)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2781..2803
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2841..2859)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2927..3031
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3032..3100,3101..3892)
                     /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(3250..3269)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4063..4651
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4552..4571
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttatgat gtgccaggct tgggagaaac accacaagca
      181 aagccaaaat aggtggccta gaacttccag cttgaaatat gggagagaat gagggaggca
      241 ctgtagagca gctgccgggt gccgcatgag aacaattctc cctgctcata attaatccta
      301 cctatttctg atgacagctg gctcttcact ttgaacaagc tagttaacaa ctttcttctc
      361 acattgagca aataattcat atttaattac ttaaccacca gttacaaaat gagaatcatc
      421 aaggaatcac aattaatttg ctattgacaa actcatactt ttagcaggct gatttctact
      481 ttatacttag attggtaatg aaaaatgaag cttattttag ttgattggtt ggacttgtgt
      541 atgaatatta tctattattt gaaaagccaa acttgaatgc aaaaaaatat tgaatatgaa
      601 aagaaaaaca tttgcagtaa agcttgttct gagctcgggc tgggcataaa agtcagggca
      661 gagccatcta ttgcttacat ttgcttctgg gatccagatc tttcgaagct agcgctaccg
      721 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
      781 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
      841 gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc
      901 tggcccaccc tcgtgaccac cctgggctac ggcgtgcagt gcttcgcccg ctaccccgac
      961 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1021 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1081 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1141 ctggggcaca agctggagta caactacaac agccacaacg tctatatcac cgccgacaag
     1201 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg
     1261 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1321 gacaaccact acctgagcta ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     1381 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1441 tacaagtaag tcgacggcgc gccgcggccg cgaattcgat atcataatca acctctggat
     1501 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     1561 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     1621 tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc cgcctgcctt
     1681 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggc tcgagagatc
     1741 ttcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     1801 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     1861 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     1921 ggattgggaa gacaatagca ggcatgagat ctcacgtgcg gaccgagcgg ccgcaggaac
     1981 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     2041 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     2101 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2161 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     2221 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc
     2281 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2341 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2401 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2461 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2521 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt
     2581 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     2641 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     2701 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     2761 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     2821 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     2881 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2941 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3001 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     3061 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     3121 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     3181 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     3241 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     3301 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     3361 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     3421 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     3481 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     3541 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     3601 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     3661 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     3721 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     3781 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3841 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3901 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3961 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4021 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4081 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4141 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4201 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4261 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4321 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4381 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4441 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4501 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4561 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4621 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4681 cctggccttt tgctggcctt ttgctcacat gt
//
