LOCUS       sequence-470524-        4667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0809m 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 4667)
  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 4667)
  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: AiP12803 - pAAV-AiE0809m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     546..565
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(784..888)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     956..974
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1012..1034)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1134..1153
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1170..1625)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1307..1328)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1519..1538
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1700..1840
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1711..1840
                     /label=AAV2 ITR
     polyA_signal    1958..2069
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2256..2276
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2369..3079,3080..3082,3083..3085))
                     /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(2411..2432)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2760..2779
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3019..3040
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3082..3091
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3640..3769
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3831..4419)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3911..3930)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4590..4667,1..714,715..783))
                     /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"
ORIGIN
        1 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
       61 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
      121 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
      181 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
      241 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
      301 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
      361 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
      421 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
      481 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
      541 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
      601 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
      661 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
      721 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
      781 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
      841 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
      901 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
      961 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     1021 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     1081 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     1141 agagcagatt gtactgagag tgcaccataa aattgtaaac gttaatattt tgttaaaatt
     1201 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa tcggcaaaat
     1261 cccttataaa tcaaaagaat agcccgagat agagttgagt gttgttccag tttggaacaa
     1321 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     1381 cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa
     1441 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     1501 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag
     1561 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     1621 cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     1681 aaataccgca tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
     1741 cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
     1801 cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgctc ggtccgcacg
     1861 tgagatctca tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc
     1921 accccccaga atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta
     1981 ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac
     2041 aacagatggc tggcaactag aaggcacagt cgaagatctc tcgagccacg gaattgtcag
     2101 tgcccaacag ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg
     2161 tggcaagaac taaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa
     2221 tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt
     2281 taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgatta tgatatcgaa
     2341 ttcgcggccg cggcgcgccg tcgacttact tgtacagctc gtccatgccg agagtgatcc
     2401 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
     2461 tcagcttgga ctggtagctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     2521 tgggggtgtt ctgctggtag tggtcggcga gctgcacgcc gccgtcctcg atgttgtggc
     2581 ggatcttgaa gttggccttg atgccgttct tctgcttgtc ggcggtgata tagacgttgt
     2641 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
     2701 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
     2761 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
     2821 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg aagcactgca
     2881 cgccgtagcc cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
     2941 agatcagctt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc
     3001 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
     3061 acagctcctc gcccttgctc accatggtgg cgaccggtag cgctagcttc gaaagatctg
     3121 gatcccagaa gcaaatgtaa gcaatagatg gctctgccct gacttttatg cccagcccga
     3181 gctcgttcat gacagatgtt agagaggccc ctgggactcc atgtcctcag attgctatag
     3241 taatggccat catacccagt cccagctgca atctgttgct ggcagggttt gtgctgtagc
     3301 tgtggccact tgcttcctgt cattaaggca aaccaccatc ttattttgtt ttgttttccc
     3361 ctcatgtttt gagtttgctg ctctccagca cttgttatta gaacagacat aattaataac
     3421 tcctggcacg tggagcagcc agattggaca atctgcctcc tgccttctca aacagtcctc
     3481 tgagcaatct gaaagcctga atttatagcc tggaccgaat acaatggatt gttttgtaat
     3541 cctaagcagg catcataaac tcactacact gacagatgaa gaaataaaat tctctttgta
     3601 gttcctagtg gatggtttac tcaacacgcg tgcggccgca ggaaccccta gtgatggagt
     3661 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     3721 gacgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcagga catgtgagca
     3781 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3841 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3901 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3961 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4021 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4081 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4141 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4201 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4261 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4321 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4381 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4441 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4501 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4561 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4621 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgt
//
