LOCUS       sequence-432580-        4723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2582m 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 4723)
  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 4723)
  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: AiP1894 - pAAV-AiE2582m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     68..87
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   249..389
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   260..389
                     /label=AAV2 ITR
     polyA_signal    507..618
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     805..825
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(918..1628,1629..1631,1632..1634))
                     /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(960..981)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1309..1328
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1568..1589
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1631..1640
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1786..1795
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2245..2374
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2436..3024)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2516..2535)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3195..3986,3987..4055))
                     /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     3818..3837
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4056..4160)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4228..4246
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4284..4306)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4406..4425
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4442..4723,1..174))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4579..4600)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
       61 gaaaggaagg gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca
      121 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg
      181 gttgctttga cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
      241 caggcgcccc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
      301 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      361 agtggccaac tccatcacta ggggttcctg cggccgctcg gtccgcacgt gagatctcat
      421 gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa
      481 tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag
      541 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
      601 ggcaactaga aggcacagtc gaagatctct cgagccacgg aattgtcagt gcccaacagc
      661 cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt ggcaagaact
      721 aaccaggatt tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac
      781 aaaggcatta aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca
      841 gtcaatcttt cacaaatttt gtaatccaga ggttgattat gatatcgaat tcgcggccgc
      901 ggcgcgccgt cgacttactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc
      961 acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagcttggac
     1021 tggtagctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc
     1081 tgctggtagt ggtcggcgag ctgcacgccg ccgtcctcga tgttgtggcg gatcttgaag
     1141 ttggccttga tgccgttctt ctgcttgtcg gcggtgatat agacgttgtg gctgttgtag
     1201 ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc
     1261 tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg
     1321 ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg
     1381 aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga agcactgcac gccgtagccc
     1441 agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatcagcttc
     1501 agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg
     1561 ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg
     1621 cccttgctca ccatggtggc gaccggtagc gctagcttcg aaagatctgg atcccagaag
     1681 caaatgtaag caatagatgg ctctgccctg acttttatgc ccagcccgag ctccgagggt
     1741 gcagagttgc ttcttggcag caccttgcct agaaccgctc tctctccatg gtggcttctc
     1801 acaggactag tcaccagtac catttgggaa agggtgggtg atggctaaac ttctctctgc
     1861 ttggccttct tcccaggagg atactggaag tgggagtgtg atgggtgaga atggggagga
     1921 ctggaatcta actgcctgtc agctccccaa aggttgggct ttgctcaaat gtcaccaaag
     1981 tgggaaccaa atgcaaggcc tgcaaactac cacagagata atgagagact cagaaacaag
     2041 attcgtcaaa agactggagg aactggaatg atttatccta aggaagggaa ggctcagcaa
     2101 cacatactgg gggcagttat gactcagtgg gaagactcaa ttctgctgat tgctaagtgc
     2161 tcacacatga atccagccct gtgcacactc agtttcttat cttaacagta accttaagga
     2221 gggacgtttt accattgcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     2281 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggcggcc
     2341 tcagtgagcg agcgagcgcg cagctgcctg caggacatgt gagcaaaagg ccagcaaaag
     2401 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2461 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2521 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2581 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2641 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2701 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2761 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2821 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2881 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2941 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3001 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3061 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3121 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3181 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3241 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3301 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3361 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3421 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3481 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3541 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3601 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3661 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3721 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3781 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3841 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3901 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3961 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4021 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4081 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4141 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     4201 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     4261 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     4321 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     4381 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     4441 aaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     4501 ttttttaacc aatagaccga aatcggcaaa atcccttata aatcaaaaga atagcccgag
     4561 atagagttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     4621 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     4681 aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa atc
//
