LOCUS       sequence-466196-        4809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2490m 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 4809)
  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 4809)
  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: AiP1947 - pAAV-AiE2490m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(573..1364,1365..1433))
                     /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     1196..1215
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1434..1538)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1606..1624
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1662..1684)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1784..1803
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1820..2275)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1957..1978)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2169..2188
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2350..2490
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2361..2490
                     /label=AAV2 ITR
     polyA_signal    2608..2719
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2906..2926
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3019..3729,3730..3732,3733..3735))
                     /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(3061..3082)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3410..3429
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3669..3690
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3732..3741
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4432..4561
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4623..4809,1..402))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4703..4722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
       61 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      121 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      181 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      241 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      301 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      361 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      421 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      481 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      541 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
      601 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
      661 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
      721 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
      781 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
      841 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
      901 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
      961 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1021 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1081 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1141 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1201 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1261 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1321 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1381 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1441 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1501 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1561 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     1621 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     1681 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     1741 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     1801 gtactgagag tgcaccataa aattgtaaac gttaatattt tgttaaaatt cgcgttaaat
     1861 ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa tcggcaaaat cccttataaa
     1921 tcaaaagaat agcccgagat agagttgagt gttgttccag tttggaacaa gagtccacta
     1981 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     2041 ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     2101 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     2161 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag tgtagcggtc
     2221 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtactat
     2281 ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2341 tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc
     2401 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     2461 gagtggccaa ctccatcact aggggttcct gcggccgctc ggtccgcacg tgagatctca
     2521 tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga
     2581 atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca
     2641 gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc
     2701 tggcaactag aaggcacagt cgaagatctc tcgagccacg gaattgtcag tgcccaacag
     2761 ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaagaac
     2821 taaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa tagcatgata
     2881 caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt taagaatacc
     2941 agtcaatctt tcacaaattt tgtaatccag aggttgatta tgatatcgaa ttcgcggccg
     3001 cggcgcgccg tcgacttact tgtacagctc gtccatgccg agagtgatcc cggcggcggt
     3061 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga
     3121 ctggtagctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     3181 ctgctggtag tggtcggcga gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa
     3241 gttggccttg atgccgttct tctgcttgtc ggcggtgata tagacgttgt ggctgttgta
     3301 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
     3361 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
     3421 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
     3481 gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg aagcactgca cgccgtagcc
     3541 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatcagctt
     3601 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg
     3661 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
     3721 gcccttgctc accatggtgg cgaccggtag cgctagcttc gaaagatctg gatcccagaa
     3781 gcaaatgtaa gcaatagatg gctctgccct gacttttatg cccagcccga gctccggaaa
     3841 gcaatcaccg aaaaagtaat tacggggcat gggggagcat ttagcggaag tcagcactaa
     3901 gtcttcactt acggacctgg tttcttggtc aaatggcaac tcgtttacct ttagatcaag
     3961 tcatttctga ataatcaacc agagctagaa atctgtagat gacaacctag gtctgatgcg
     4021 ttctgacagc acagatggct gccgtctgat ttgcatacta acaaagctct agtgtaattg
     4081 agtaatgcac cagggggaag gtggcatcat tcccaaagaa gggaaatgtg atttctaaaa
     4141 ccacttggca ccaaccaagt gacacttttc tcaggctcat ttaaaagtct attacagggc
     4201 gtggggagat ggtacaatgg gaaaagtgct gctggccggg caagcctgag gacttgggtt
     4261 tggatcctca gaactccgcg taaagccaga cacagcagca tgcgtatata aaccatagct
     4321 tcaaaggcaa aaggggaggc ggacacagag accttaagat aacgggccag ctagtgtggc
     4381 attatgcagt tgcaaacaag aaactatctc aaacaaggcg gatgcggccg caggaacccc
     4441 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     4501 caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     4561 gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4621 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4681 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4741 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4801 agcgtggcg
//
