LOCUS       sequence-466348-        4765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3055m 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 4765)
  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 4765)
  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: AiP20018 - pAAV-AiE3055m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      277..286
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(283..285,286..288,289..999)
                     /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(328..349)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(589..608)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     936..957
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1092..1112)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1299..1410
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1528..1668
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1528..1657
                     /label=AAV2 ITR
     rep_origin      1743..2198
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1830..1849)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2040..2061
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2215..2234)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2334..2356
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2394..2412)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2480..2584
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2585..2653,2654..3445)
                     /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(2803..2822)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3616..4204
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4105..4124
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4266..4395
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 atgttacaat atcctgtact actttataaa gaaaagctaa tttcggtcac agttctgaag
       61 cctggaagcc ggatatttta tgtcttcatg tgtggcagca ggtgagaaca tcatgacata
      121 cggtatcaca gcagcagtaa tacttgcaaa gagatcacag gaggagatgg gaagccgccg
      181 gcggagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc
      241 tgggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtgag caagggcgag
      301 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      361 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      421 ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc
      481 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
      541 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
      601 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
      661 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
      721 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
      781 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
      841 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
      901 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
      961 gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg
     1021 ccgcgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     1081 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     1141 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt
     1201 cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     1261 ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc
     1321 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     1381 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1441 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga
     1501 gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     1561 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     1621 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     1681 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     1741 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1801 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     1861 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     1921 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     1981 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2041 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     2101 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     2161 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     2221 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     2281 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     2341 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     2401 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     2461 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2521 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2581 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2641 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2701 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2761 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2821 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2881 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2941 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3001 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3061 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3121 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3181 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3241 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3301 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3361 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3421 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3481 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3541 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3601 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3661 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3721 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3781 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3841 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3901 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3961 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4021 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4081 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4141 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4201 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4261 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
     4321 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
     4381 tcactagggg ttcctgcggc cgcacgcgta cagatattta caacccaact gctaacagta
     4441 gcaaaaatta cagttatgga gtagcaacaa aaacaaccat aaagcagtaa ttttatggct
     4501 gagagtcacc acagcatgag caacattgca gcattagaaa ggatgagaac tactgttcta
     4561 ggcatatggc tttcaaagga gtgcacatca gaattacata cagctttccc agaatgcacc
     4621 tgagccaggt cctatcccag gcactcagtt tagtggtgta gggtgagatt ggaatttgag
     4681 taaattgtag cagttactca agtgactcta aaatgcaggc ctagtatgaa tacattccta
     4741 gaggcttggt tgtggattct ctgtc
//
