LOCUS       sequence-438965-        4746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2569m 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 4746)
  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 4746)
  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: AiP1976 - pAAV-AiE2569m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(223..1014,1015..1083))
                     /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     846..865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1084..1188)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1256..1274
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1312..1334)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1434..1453
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1470..1925)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1607..1628)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1819..1838
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2000..2140
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2011..2140
                     /label=AAV2 ITR
     polyA_signal    2258..2369
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2556..2576
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2669..3379,3380..3382,3383..3385))
                     /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(2711..2732)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3060..3079
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3319..3340
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3382..3391
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4019..4148
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4210..4746,1..52))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4290..4309)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
       61 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      121 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      181 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      241 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
      301 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
      361 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
      421 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
      481 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
      541 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
      601 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
      661 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
      721 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
      781 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
      841 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
      901 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
      961 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1021 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1081 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1141 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1201 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     1261 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     1321 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     1381 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     1441 agagcagatt gtactgagag tgcaccataa aattgtaaac gttaatattt tgttaaaatt
     1501 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa tcggcaaaat
     1561 cccttataaa tcaaaagaat agcccgagat agagttgagt gttgttccag tttggaacaa
     1621 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     1681 cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa
     1741 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     1801 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag
     1861 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     1921 cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     1981 aaataccgca tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
     2041 cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
     2101 cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgctc ggtccgcacg
     2161 tgagatctca tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc
     2221 accccccaga atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta
     2281 ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac
     2341 aacagatggc tggcaactag aaggcacagt cgaagatctc tcgagccacg gaattgtcag
     2401 tgcccaacag ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg
     2461 tggcaagaac taaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa
     2521 tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt
     2581 taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgatta tgatatcgaa
     2641 ttcgcggccg cggcgcgccg tcgacttact tgtacagctc gtccatgccg agagtgatcc
     2701 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
     2761 tcagcttgga ctggtagctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     2821 tgggggtgtt ctgctggtag tggtcggcga gctgcacgcc gccgtcctcg atgttgtggc
     2881 ggatcttgaa gttggccttg atgccgttct tctgcttgtc ggcggtgata tagacgttgt
     2941 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
     3001 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
     3061 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
     3121 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg aagcactgca
     3181 cgccgtagcc cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
     3241 agatcagctt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc
     3301 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
     3361 acagctcctc gcccttgctc accatggtgg cgaccggtag cgctagcttc gaaagatctg
     3421 gatcccagaa gcaaatgtaa gcaatagatg gctctgccct gacttttatg cccagcccga
     3481 gctccgaaac atcgagcact gcttggtagc acttcagttc caccttggct tattggttac
     3541 ttttttctca ctttgtgcag aaaagcaact taaggaagga aggacttctt ctggcttgca
     3601 gttggagagt tagtccgtcg tgacaagtta gactacagca gctcaattag agcgttggcg
     3661 acagaaagaa gtgtgtgttg tctggtcaca gagacatatc tcctaggtga ttctaaatcc
     3721 agttatgatg acaatcaaga gtcaccatca cacatggtta taattgtata tacacataaa
     3781 tatgtacata tatatatata cttccacacc tacgtgcaca tgtatttata cttttaatcc
     3841 tcttgattta gtaagaactc ctctgctttg aagtaacaga gccagcactg gcaatgtgac
     3901 actgagtgat gtgagcttga tccctagtat tcctatggag gaaggagaga atggatagaa
     3961 tggatttcta ctcatggtcc tctgcctctc acatacaggc cataataact gcggccgcag
     4021 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     4081 gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     4141 cctgcaggac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4201 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4261 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4321 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4381 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4441 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4501 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4561 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4621 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4681 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4741 tagcgg
//
