LOCUS       sequence-466306-        4513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2520m 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 4513)
  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 4513)
  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: AiP1784 - pAAV-AiE2520m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(332..1123,1124..1192))
                     /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     955..974
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1193..1297)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1365..1383
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1421..1443)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1543..1562
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1579..2034)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1716..1737)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1928..1947
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2109..2249
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2120..2249
                     /label=AAV2 ITR
     polyA_signal    2367..2478
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2665..2685
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2778..3488,3489..3491,3492..3494))
                     /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(2820..2841)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3169..3188
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3428..3449
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3491..3500
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3895..4024
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4086..4513,1..161))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4166..4185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
       61 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
      121 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
      181 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
      241 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
      301 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
      361 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
      421 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
      481 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
      541 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
      601 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
      661 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
      721 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
      781 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
      841 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
      901 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
      961 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     1021 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     1081 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     1141 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     1201 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     1261 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     1321 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     1381 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     1441 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     1501 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     1561 tactgagagt gcaccataaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt
     1621 tttgttaaat cagctcattt tttaaccaat agaccgaaat cggcaaaatc ccttataaat
     1681 caaaagaata gcccgagata gagttgagtg ttgttccagt ttggaacaag agtccactat
     1741 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     1801 tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     1861 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     1921 gaaaggaagg gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca
     1981 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg
     2041 gttgctttga cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     2101 caggcgcccc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
     2161 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
     2221 agtggccaac tccatcacta ggggttcctg cggccgctcg gtccgcacgt gagatctcat
     2281 gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa
     2341 tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag
     2401 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
     2461 ggcaactaga aggcacagtc gaagatctct cgagccacgg aattgtcagt gcccaacagc
     2521 cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt ggcaagaact
     2581 aaccaggatt tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac
     2641 aaaggcatta aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca
     2701 gtcaatcttt cacaaatttt gtaatccaga ggttgattat gatatcgaat tcgcggccgc
     2761 ggcgcgccgt cgacttactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc
     2821 acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagcttggac
     2881 tggtagctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc
     2941 tgctggtagt ggtcggcgag ctgcacgccg ccgtcctcga tgttgtggcg gatcttgaag
     3001 ttggccttga tgccgttctt ctgcttgtcg gcggtgatat agacgttgtg gctgttgtag
     3061 ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc
     3121 tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg
     3181 ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg
     3241 aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga agcactgcac gccgtagccc
     3301 agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatcagcttc
     3361 agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg
     3421 ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg
     3481 cccttgctca ccatggtggc gaccggtagc gctagcttcg aaagatctgg atcccagaag
     3541 caaatgtaag caatagatgg ctctgccctg acttttatgc ccagcccgag ctccgaagca
     3601 tcagaagagc ctgaaaggct tgtttctgtg agaacacaac tgttcactgc tgacaatgtc
     3661 tctaacccct cggaaaggga cagcatgaag cctgtacatt taatttgctc ccttaactct
     3721 aggagggaag ggagtcacag acagaatgaa tacaaaaata gttatgttaa tgatcccttg
     3781 agtttgttcc tcggtttctc aagaaacaag aaacgaagtt ctgtttatag ctaacaaaag
     3841 tgatggagga gattatttca gggcgattat tactcttaca acgcgtgcgg ccgcaggaac
     3901 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     3961 gaccaaaggt cgcccgacgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     4021 caggacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4081 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4141 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4201 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4261 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4321 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4381 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4441 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4501 tggcctaact acg
//
