LOCUS       sequence-438944-        5292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2425m 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 5292)
  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 5292)
  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: AiP1878 - pAAV-AiE2425m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..134
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(135..203,204..995)
                     /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(353..372)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1166..1754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1655..1674
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1816..1945
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3119..3128
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3125..3127,3128..3130,3131..3841)
                     /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(3170..3191)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3431..3450)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3778..3799
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3934..3954)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4141..4252
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4370..4510
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4370..4499
                     /label=AAV2 ITR
     rep_origin      4585..5040
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4672..4691)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4882..4903
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5057..5076)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5176..5198
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5236..5254)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
       61 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
      121 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
      181 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
      241 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
      301 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
      361 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
      421 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
      481 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
      541 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
      601 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
      661 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
      721 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
      781 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
      841 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
      901 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
      961 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     1021 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     1081 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     1141 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     1201 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1261 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     1321 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1381 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1441 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1501 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     1561 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     1621 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     1681 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     1741 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     1801 cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
     1861 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
     1921 gccaactcca tcactagggg ttcctgcggc cgcacgcgtt gtaagagtaa taatcgggcc
     1981 ctcgtctcta aacccctcta ttctgctttt tccagtagaa gtaaggcctc cctgggcttc
     2041 cccaaaatgc agacatatac atgctcagcc taaaattcag gaacaagcga tcagtcctcc
     2101 taacaccacc gtccctcaga atctgaggag gtacggatga gtgtacacaa tctgtaacct
     2161 aagcatttcc ttctaaatat ttatctgatg catttttgaa tattttgaaa ggcctttagt
     2221 ctcccattcc attgtaagat caaatgttcc tatttttctc tattcaaagg agttttcact
     2281 attcacacat cacagggaaa tcgatgaact cattcaccat tcccaccacc ctccctccac
     2341 cttctcccca cctgccttcc aatatacatt tacatccatt tcatgcaatt ctctcccttc
     2401 ctctcctccc tcctccccca ttttgtcatt tcccccttta aatggtagaa attgcaagct
     2461 ttcctttctt gacagttatg attaagggac tctgcacctg ctcaggtgag tgaatgggag
     2521 agacgctgaa gcacatactt atgctaacta ataccttagt ctccttccat gtctgcaaaa
     2581 aaaggtttgc agctcagagg gaccgcccct taagtaagtg cattaagcat ttcctgttca
     2641 agttcctcac gaagtcaaaa gagctctttg caccttgact catgatgcat gttagggcgc
     2701 aggggcgctt tctgtagaga tgaatgggtg ctcagctcac gtggaaagca agaaggaaat
     2761 gagaaacgca ggagagaggg caggaagacg tagcaggctt tgatgggaaa tggcttagcc
     2821 cttccccctt ggcataaact tgaggggccc taagactaaa agcgggcaca ctctatagta
     2881 gagtgcccgt gatctccatg ttgctgtctc atttccaaag tcatctccct ctccacttag
     2941 aattcacagc tatttttcct tctctgatgg gaaaaggcag tcttgctaga catgtaccca
     3001 attagtatgt aaattttact gaacggagct cgggctgggc ataaaagtca gggcagagcc
     3061 atctattgct tacatttgct tctgggatcc agatctttcg aagctagcgc taccggtcgc
     3121 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     3181 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     3241 ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc
     3301 caccctcgtg accaccctgg gctacggcgt gcagtgcttc gcccgctacc ccgaccacat
     3361 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     3421 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     3481 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3541 gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa
     3601 gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct
     3661 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3721 ccactacctg agctaccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     3781 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     3841 gtaagtcgac ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc tggattacaa
     3901 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3961 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     4021 cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg
     4081 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggctcgag agatcttcga
     4141 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     4201 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     4261 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     4321 gggaagacaa tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta
     4381 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     4441 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     4501 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4561 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4621 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     4681 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4741 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4801 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4861 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     4921 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     4981 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     5041 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     5101 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     5161 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     5221 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     5281 tggtttctta ga
//
