LOCUS       sequence-466314-        4666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2559m 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 4666)
  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 4666)
  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: AiP1866 - pAAV-AiE2559m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..22
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   184..324
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   195..324
                     /label=AAV2 ITR
     polyA_signal    442..553
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     740..760
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(853..1563,1564..1566,1567..1569))
                     /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(895..916)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1244..1263
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1503..1524
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1566..1575
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2123..2252
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2314..2902)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2394..2413)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3073..3864,3865..3933))
                     /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     3696..3715
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3934..4038)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4106..4124
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4162..4184)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4284..4303
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4320..4666,1..109))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4457..4478)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg
       61 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta ctatggttgc
      121 tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
      181 gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc
      241 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
      301 ccaactccat cactaggggt tcctgcggcc gctcggtccg cacgtgagat ctcatgcctg
      361 ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc cagaatagaa
      421 tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag gacagtggga
      481 gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga tggctggcaa
      541 ctagaaggca cagtcgaaga tctctcgagc cacggaattg tcagtgccca acagccgagc
      601 ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa gaactaacca
      661 ggatttatac aaggaggaga aaatgaaagc catacgggaa gcaatagcat gatacaaagg
      721 cattaaagca gcgtatccac atagcgtaaa aggagcaaca tagttaagaa taccagtcaa
      781 tctttcacaa attttgtaat ccagaggttg attatgatat cgaattcgcg gccgcggcgc
      841 gccgtcgact tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
      901 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct tggactggta
      961 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     1021 gtagtggtcg gcgagctgca cgccgccgtc ctcgatgttg tggcggatct tgaagttggc
     1081 cttgatgccg ttcttctgct tgtcggcggt gatatagacg ttgtggctgt tgtagttgta
     1141 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     1201 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     1261 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     1321 gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac tgcacgccgt agcccagggt
     1381 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatca gcttcagggt
     1441 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     1501 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     1561 gctcaccatg gtggcgaccg gtagcgctag cttcgaaaga tctggatccc agaagcaaat
     1621 gtaagcaata gatggctctg ccctgacttt tatgcccagc ccgagctccg agttggctgg
     1681 tttggggtaa atgatgctga actatttgca aaaggcaaac aatggttcag gctgctttct
     1741 actgtatctt cccatagctt gccaggatct gaggttccat tgcctagcct ctccaggtcg
     1801 gcagtgcaca gataactgtt gtaaaattgg catactccac cacactcagt agcagcaaga
     1861 tgtgagacta gatacagagc ctttccttac cacacggtgg cacaacacca agcccatggt
     1921 gtcctgtggc agccacaggg cggtattgtg aggtctggga cctttgagga cgggctgcag
     1981 ccccaggagg cgatacatga tctctgctgg gcagggccca ggcaaagcca gcccactgtg
     2041 gggaccgtgt cctcctggga tcccccagta gaagcctagg gaagacacac cctagcaggc
     2101 tatctttaag taatgcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     2161 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggcggcctc
     2221 agtgagcgag cgagcgcgca gctgcctgca ggacatgtga gcaaaaggcc agcaaaaggc
     2281 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2341 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2401 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2461 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2521 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2581 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2641 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2701 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2761 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2821 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2881 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2941 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3001 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3061 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3121 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3181 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3241 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3301 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3361 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3421 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3481 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3541 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3601 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3661 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3721 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3781 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3841 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3901 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3961 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4021 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     4081 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     4141 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     4201 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     4261 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa
     4321 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     4381 ttttaaccaa tagaccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat
     4441 agagttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     4501 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa
     4561 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     4621 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaag
//
