LOCUS       sequence-466322-        4656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2571m 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 4656)
  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 4656)
  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: AiP1884 - pAAV-AiE2571m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4656
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(533..1324,1325..1393))
                     /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     1156..1175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1394..1498)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1566..1584
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1622..1644)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1744..1763
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1780..2235)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1917..1938)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2129..2148
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2310..2450
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2321..2450
                     /label=AAV2 ITR
     polyA_signal    2568..2679
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2866..2886
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2979..3689,3690..3692,3693..3695))
                     /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(3021..3042)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3370..3389
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3629..3650
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3692..3701
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4239..4368
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4430..4656,1..362))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4510..4529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
       61 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      121 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      181 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
      241 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      301 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      361 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      421 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
      481 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
      541 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
      601 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
      661 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
      721 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
      781 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
      841 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
      901 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
      961 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1021 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1081 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1141 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1201 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1261 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1321 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1381 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     1441 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     1501 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     1561 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     1621 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     1681 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     1741 atgcggcatc agagcagatt gtactgagag tgcaccataa aattgtaaac gttaatattt
     1801 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa
     1861 tcggcaaaat cccttataaa tcaaaagaat agcccgagat agagttgagt gttgttccag
     1921 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     1981 tctatcaggg cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga
     2041 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     2101 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg
     2161 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     2221 cgctacaggg cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg
     2281 cgtaaggaga aaataccgca tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac
     2341 tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
     2401 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgctc
     2461 ggtccgcacg tgagatctca tgcctgctat tgtcttccca atcctccccc ttgctgtcct
     2521 gccccacccc accccccaga atagaatgac acctactcag acaatgcgat gcaatttcct
     2581 cattttatta ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa ggcacggggg
     2641 aggggcaaac aacagatggc tggcaactag aaggcacagt cgaagatctc tcgagccacg
     2701 gaattgtcag tgcccaacag ccgagcccct gtccagcagc gggcaaggca ggcggcgatg
     2761 agttccgccg tggcaagaac taaccaggat ttatacaagg aggagaaaat gaaagccata
     2821 cgggaagcaa tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga
     2881 gcaacatagt taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgatta
     2941 tgatatcgaa ttcgcggccg cggcgcgccg tcgacttact tgtacagctc gtccatgccg
     3001 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
     3061 gggtctttgc tcagcttgga ctggtagctc aggtagtggt tgtcgggcag cagcacgggg
     3121 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgcc gccgtcctcg
     3181 atgttgtggc ggatcttgaa gttggccttg atgccgttct tctgcttgtc ggcggtgata
     3241 tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc
     3301 ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc
     3361 tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag
     3421 ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg
     3481 aagcactgca cgccgtagcc cagggtggtc acgagggtgg gccagggcac gggcagcttg
     3541 ccggtggtgc agatcagctt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg
     3601 ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc
     3661 accccggtga acagctcctc gcccttgctc accatggtgg cgaccggtag cgctagcttc
     3721 gaaagatctg gatcccagaa gcaaatgtaa gcaatagatg gctctgccct gacttttatg
     3781 cccagcccga gctccgcctg aggagtatca gtgacccagc ctagtgtagg ctctctgagc
     3841 cgtgagcctt ggagattaca gccctggtca gagtccaggt gtatctgtgt gctatacctg
     3901 ttcacatgtt agtgtacaca tgtgaagatg tttctgtaca tgtgtgtata tgcatatctt
     3961 tgtatctata tagtttattt gtgtatattt gcctatagtc tgtttctgtg tacacaggac
     4021 catgcatatt ggtatctgtt tctgtgagtg gttgtgtgtc tatgcctatt tgtctctgaa
     4081 ggcgtttgta agtgtccatg catgtgtgtg cgcatgaact catgtgtata tatgtgtacc
     4141 tttgtctgtg tgtccatgaa tacacattca tgtatgtgtc tacacatgtg tacctatata
     4201 tctatgtata aatgcatgtg tgtctatatt gcggccgcag gaacccctag tgatggagtt
     4261 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     4321 acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcaggac atgtgagcaa
     4381 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4441 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4501 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4561 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4621 ctcatagctc acgctgtagg tatctcagtt cggtgt
//
