LOCUS       sequence-462007-        4870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2476m 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 4870)
  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 4870)
  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: AiP1864 - pAAV-AiE2476m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4870
                     /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      882..891
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(888..890,891..893,894..1604)
                     /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(933..954)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1194..1213)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1541..1562
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1697..1717)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1904..2015
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2133..2273
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2133..2262
                     /label=AAV2 ITR
     rep_origin      2348..2803
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2435..2454)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2645..2666
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2820..2839)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2939..2961
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2999..3017)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3085..3189
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3190..3258,3259..4050)
                     /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(3408..3427)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4221..4809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4710..4729
                     /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 gcgttgtaag agtaataatc gagaatggta cagctgcaag
      181 cctcaacacc cagcatgcac cgcagctggg gagtgctggg agcctttgaa gagcccgggc
      241 aggatgccac aggcatctgc gggaagcagg ggatacacaa ggcagggtca gcactgactg
      301 atttcaacca ctacaaggca tgtagccatc tcttccctct ttctcttctt ccttctagat
      361 cttgtcttcc tgcgccctca ttgctcgcgt ttctgattca tgtatctgac caccatcagc
      421 agatctgaag aggacaggaa gcacccagtt cgaactcttc caagtcgggg cctgcacacc
      481 ccacccccac ccccttcctg gagtctgtct gggaggggat ccccaatgac gcactcggga
      541 aggcagtaca gagccccacg ccaggcccgg ggatcatgaa tgagggcaga gagaaggcca
      601 ggcgcttcct cctcaccagc cggagctttg gcccccagca gggtgagcag ctgccctcct
      661 gaggcagccc aggcaggctc tatgcccagc ccttgttggc cagaagacaa gctggctggg
      721 tccccgcgaa agaggtcgtt tcaatagctt tcaccctgca ctcccgcaca ccaccccagc
      781 cccggccgga gctcgggctg ggcataaaag tcagggcaga gccatctatt gcttacattt
      841 gcttctggga tccagatctt tcgaagctag cgctaccggt cgccaccatg gtgagcaagg
      901 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
      961 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1021 tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     1081 tgggctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     1141 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1201 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1261 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1321 actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca
     1381 acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc
     1441 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     1501 agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     1561 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagtc gacggcgcgc
     1621 cgcggccgcg aattcgatat cataatcaac ctctggatta caaaatttgt gaaagattga
     1681 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     1741 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     1801 tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     1861 ggctgttggg cactgacaat tccgtggctc gagagatctt cgactgtgcc ttctagttgc
     1921 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     1981 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2041 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2101 catgagatct cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     2161 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     2221 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     2281 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     2341 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2401 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     2461 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2521 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     2581 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2641 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     2701 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     2761 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     2821 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     2881 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     2941 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     3001 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     3061 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3121 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3181 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3241 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3301 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3361 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3421 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3481 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3541 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3601 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3661 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3721 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3781 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3841 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3901 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3961 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4021 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4081 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4141 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4201 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4261 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4321 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4381 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4441 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4501 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4561 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4621 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4681 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4741 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4801 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4861 gctcacatgt
//
