LOCUS       sequence-473572-        6342 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0825m 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 6342)
  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 6342)
  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: AiP12895 - pAAV-AiE0825m-SYFP2-WPRE3-BGHpA (Al...
FEATURES             Location/Qualifiers
     source          1..6342
                     /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      2354..2363
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2360..2362,2363..2365,2366..3076)
                     /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(2405..2426)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2666..2685)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3013..3034
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3169..3189)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3376..3487
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3605..3745
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3605..3734
                     /label=AAV2 ITR
     rep_origin      3820..4275
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3907..3926)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4117..4138
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4292..4311)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4411..4433
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4471..4489)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4557..4661
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4662..4730,4731..5522)
                     /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(4880..4899)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5693..6281
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6182..6201
                     /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 gcgtaccggc cctctcctct ccccactacg gggatttctg
      181 ggtggctcac agtggacaag gaataggtaa tcccaccaaa ccttgaggtg ggctaaactg
      241 gagattttgg tatgggtctc ctactgtaaa gagaaagcct tccgggctaa cggtattcct
      301 ttcgcagtct actgcctcct ttgtgaacta atgactgtaa ttattacctc ccagagctct
      361 tttgttatca acaacccgga gccagagggg ggtgggggat gggccctttg ggaaaaggga
      421 aaaaaattta caaagcaaat tgctgtctac gaagggttgg ttttttaaaa agacaaaccc
      481 aattccagcc tgcattcgaa gtggggagtg gcaaaatttt ttttcaaagt atgataagat
      541 tttaaaaata ctcagagctg tgggattcaa ttgcatttcc acagtatcgc tttaagttac
      601 tatttttgac gaagtaaaat tcagatgttg aaaatctgta tctcttgttt tcatccattt
      661 cttccttgga aggtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt
      721 gtgtgtgttt aagaaccttt tttttttatt tccgaccact catccatggt aaaaagcttc
      781 aaccccagcc agctggcttg aagagaatta ttttgcagga tttagtatgg atttaaacaa
      841 agactcaggc tctccaagtc ttgttggcaa ggctgggcgg aagcgccctt tccaggagct
      901 gttacctctg ccagctgaag ccaccaggcc tggaggaccc taaagactta ctaagttgag
      961 gcctagggtc aaggtgactg ccttccctta gagacacgcc ttgggtagca cctcagcagt
     1021 gccaatctca ccgccctccc cctccccacc tcgggccact caccggacct cagtgggcat
     1081 cctctagcta gatcctccct ggcccccctg gattctggaa accttgggaa gaagataaag
     1141 ctttgtggag agaatgcact tggtcatctg cagttattcg aggtctccgg atccctaaca
     1201 aaataagtga tcctggtcac caagactaaa ggtcccaggg tttctcctcc ggcaacagct
     1261 ttggtgactc atttgaaaaa ctcaaagaag gaataaagga ctatggagag cagggaagct
     1321 gcagtccctc gctcggagag aaggaaagga gatcgggaac ggtttaaagc agcactttcc
     1381 tgcaagtctt tcttgagctc tctgtaagcc tggaactatg ggtttagtat aggcaagagg
     1441 aaacggaagc ggagccagag ggccccaaat cttcctcaag gcgccgtccc tgcatttaag
     1501 agcctttggg aaaagggaaa cagaaaggat tttttcattc ctttattaga aaacacaaac
     1561 gaaactactt ctttctgggc tgggaaattg ccactgttgt gttcctaggc acctgctgat
     1621 ggcaagaact cctcttggga agggacgagg tcagtgggtt gagtatccct gaaaagatgg
     1681 gagtgccttt tcctgggatt tggaaaaact gattcgctta ctttgagact ccctgaatcc
     1741 tgtgtatgtg tcagctctca cccactccaa gaagtttctg ttcgttttat ttatttaatt
     1801 tttctcagcc tgaggtcctc agtgatagat tccagcgtgg attttaattg cttcaatcag
     1861 cagcctttct cttcagccgt cagtcaaaac ctggacgatg ggtctgggga cccaggcatg
     1921 actgcacaga aaccgtacga cccccacccc caccccccag gctggctcag ctcagcaggc
     1981 agacattagg ctctgctaag gagatgaaat gagttcgttg gtgtggaagc ggtccacctt
     2041 cctccctcct tggttccttc cccaagcttt ggagctcttc ctgattgagg ccaggtggcc
     2101 tgggctcttg aggtcacagc gacctcagca ctggtttgga ggaaggcggg cgcgagaacc
     2161 acttttgctc ggcagcagcc agagacagcc ctcaaacaat gagccaatcg atgcacacgt
     2221 gaaattgccc tcggccacca ttccaatctt gccaaggtct ccctgtccct ggggtcacct
     2281 aggccagcgt gctgagggct gtggtagcac cctggacagc gcgttccggc agaagcagac
     2341 ctggagaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2401 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     2461 cgagggcgat gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct
     2521 gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggcgtgcagt gcttcgcccg
     2581 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2641 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2701 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2761 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcac
     2821 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
     2881 cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2941 gctgctgccc gacaaccact acctgagcta ccagtccaag ctgagcaaag accccaacga
     3001 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     3061 ggacgagctg tacaagtaag tcgacggcgc gccgcggccg cgaattcgat atcataatca
     3121 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3181 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3241 tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc
     3301 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggc
     3361 tcgagagatc ttcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     3421 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     3481 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     3541 gcaaggggga ggattgggaa gacaatagca ggcatgagat ctcacgtgcg gaccgagcgg
     3601 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3661 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     3721 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     3781 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     3841 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     3901 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3961 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4021 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4081 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4141 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     4201 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4261 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4321 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4381 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4441 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4501 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4561 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4621 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4681 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4741 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4801 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4861 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4921 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4981 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5041 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5101 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5161 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5221 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5281 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5341 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5401 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5461 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5521 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5581 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5641 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5701 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5761 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5821 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5881 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5941 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6001 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6061 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6121 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6181 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6241 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6301 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
