LOCUS       sequence-462012-        4802 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2478m 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 4802)
  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 4802)
  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: AiP1929 - pAAV-AiE2478m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4802
                     /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      814..823
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(820..822,823..825,826..1536)
                     /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(865..886)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1126..1145)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1473..1494
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1629..1649)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1836..1947
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2065..2205
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2065..2194
                     /label=AAV2 ITR
     rep_origin      2280..2735
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2367..2386)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2577..2598
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2752..2771)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2871..2893
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2931..2949)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3017..3121
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3122..3190,3191..3982)
                     /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(3340..3359)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4153..4741
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4642..4661
                     /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 gcggccgcaa gcaggaccca tgtgggccag ccactgtccc tctttctgta
      181 gctgactgtt gtcagctaat tcagtaaatt aacgttgatt gatcattgtc tctgatctgg
      241 acatagtgac ggggctcctg acacaaaagt gaggagtcag ggcagtttct ttaaggacat
      301 agataataac tgcaatggta tgtgttggaa gcagcaagac caatcttcat ggtttaaagt
      361 aggtgaggtg gtcagagtta cagaaagtaa gcgcttgccc aggttaccca acagacatac
      421 atacagatga tttgaatcag tttgtgacct taaccaccac actgtagagt caggtgaaga
      481 atgaaggcaa ataggttgga gataagagac ctagggatgg ggactgtgat taggagctgt
      541 gagagatact agaaggactg tcccctgtca cagagggcag gcaagaagcc acatttgaca
      601 gaaaggctgt cacaccaccc atctgaaagc aagcactggt tctcagcagg aagtcaggct
      661 ttgtttcaac tctcccatct ttgggagatc tgttcagtgt cttgttctct ctcacactcg
      721 gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg
      781 gatccagatc tttcgaagct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag
      841 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      901 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg
      961 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac
     1021 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1081 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1141 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1201 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1261 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     1321 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     1381 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag
     1441 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1501 gccgggatca ctctcggcat ggacgagctg tacaagtaag tcgacggcgc gccgcggccg
     1561 cgaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     1621 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     1681 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt
     1741 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     1801 ggcactgaca attccgtggc tcgagagatc ttcgactgtg ccttctagtt gccagccatc
     1861 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     1921 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     1981 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat
     2041 ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     2101 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     2161 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     2221 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     2281 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2341 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2401 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2461 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     2521 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2581 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     2641 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2701 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     2761 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     2821 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     2881 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     2941 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     3001 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3061 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3121 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     3181 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     3241 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3301 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3361 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3421 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3481 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3541 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3601 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3661 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3721 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3781 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3841 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3901 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3961 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     4021 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4081 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4141 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4201 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4261 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4321 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4381 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4441 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4501 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4561 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4621 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4681 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4741 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4801 gt
//
