LOCUS       sequence-466343-        4649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2601m 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 4649)
  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 4649)
  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: AiP2053 - pAAV-AiE2601m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      347..356
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(353..355,356..358,359..1069)
                     /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(398..419)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(659..678)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1006..1027
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1162..1182)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1369..1480
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1598..1738
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1598..1727
                     /label=AAV2 ITR
     rep_origin      1813..2268
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1900..1919)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2110..2131
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2285..2304)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2404..2426
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2464..2482)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2550..2654
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2655..2723,2724..3515)
                     /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(2873..2892)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3686..4274
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4175..4194
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4336..4465
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gttgggaaac tgactctctc tccagatggc ttcagtgact cattgcaaag ttacgagtaa
       61 agatagcacg gtatgtgaaa cactcgggta cagcacgcac taagtgtcac cggtgacacc
      121 taggactttg cctggcaggg agtggacact aaatattgtt ggatgaatga acgtgactct
      181 ctcttgtcaa gctcctgcac ttcaagcact ggagaatggg agtacatatt gtttatacag
      241 ctactgtgga gcggagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta
      301 catttgcttc tgggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtgag
      361 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      421 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      481 gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
      541 caccctgggc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
      601 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
      661 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
      721 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
      781 gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
      841 ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta
      901 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
      961 ctaccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1021 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacgg
     1081 cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag
     1141 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     1201 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     1261 ctggttagtt cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     1321 ggctcggctg ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta
     1381 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     1441 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     1501 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     1561 gcaggcatga gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     1621 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     1681 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     1741 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     1801 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1861 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     1921 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     1981 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     2041 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2101 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     2161 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     2221 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     2281 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     2341 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     2401 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     2461 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     2521 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2581 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2641 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2701 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2761 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2821 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2881 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2941 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3001 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3061 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3121 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3181 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3241 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3301 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3361 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3421 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3481 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3541 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3601 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3661 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3721 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3781 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3841 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3901 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3961 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4021 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4081 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4141 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4201 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4261 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4321 cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
     4381 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
     4441 gccaactcca tcactagggg ttcctgcggc cgcacgcgtg cagggaaggg tgggtggaaa
     4501 aagggggatg ctgacgctga gatgggaagg ggagatagga aaacttggtc cgaaggtgtc
     4561 catccgggtg gccccaccct acccaatcta gtgttctggc tggcagagag tggatgccta
     4621 atgtggtcag cccctaagaa aaggctgca
//
