LOCUS       sequence-432559-        4829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2449m 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 4829)
  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 4829)
  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: AiP1774 - pAAV-AiE2449m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(295..1086,1087..1155))
                     /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     918..937
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1156..1260)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1328..1346
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1384..1406)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1506..1525
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1542..1997)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1679..1700)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1891..1910
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2072..2212
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2083..2212
                     /label=AAV2 ITR
     polyA_signal    2330..2441
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2628..2648
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2741..3451,3452..3454,3455..3457))
                     /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(2783..2804)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3132..3151
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3391..3412
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3454..3463
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4174..4303
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4365..4829,1..124))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4445..4464)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
       61 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      121 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      181 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      241 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
      301 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
      361 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
      421 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
      481 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
      541 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
      601 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
      661 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
      721 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
      781 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
      841 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
      901 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
      961 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1021 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1081 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1141 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     1201 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1261 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     1321 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     1381 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     1441 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     1501 ctatgcggca tcagagcaga ttgtactgag agtgcaccat aaaattgtaa acgttaatat
     1561 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aatagaccga
     1621 aatcggcaaa atcccttata aatcaaaaga atagcccgag atagagttga gtgttgttcc
     1681 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     1741 cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc
     1801 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     1861 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctaa
     1921 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     1981 gccgctacag ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat accgcacaga
     2041 tgcgtaagga gaaaataccg catcaggcgc ccctgcaggc agctgcgcgc tcgctcgctc
     2101 actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
     2161 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc
     2221 tcggtccgca cgtgagatct catgcctgct attgtcttcc caatcctccc ccttgctgtc
     2281 ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc
     2341 ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg
     2401 ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaagatc tctcgagcca
     2461 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
     2521 tgagttccgc cgtggcaaga actaaccagg atttatacaa ggaggagaaa atgaaagcca
     2581 tacgggaagc aatagcatga tacaaaggca ttaaagcagc gtatccacat agcgtaaaag
     2641 gagcaacata gttaagaata ccagtcaatc tttcacaaat tttgtaatcc agaggttgat
     2701 tatgatatcg aattcgcggc cgcggcgcgc cgtcgactta cttgtacagc tcgtccatgc
     2761 cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
     2821 tggggtcttt gctcagcttg gactggtagc tcaggtagtg gttgtcgggc agcagcacgg
     2881 ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg ccgccgtcct
     2941 cgatgttgtg gcggatcttg aagttggcct tgatgccgtt cttctgcttg tcggcggtga
     3001 tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct
     3061 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
     3121 cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt
     3181 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg
     3241 cgaagcactg cacgccgtag cccagggtgg tcacgagggt gggccagggc acgggcagct
     3301 tgccggtggt gcagatcagc ttcagggtca gcttgccgta ggtggcatcg ccctcgccct
     3361 cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
     3421 ccaccccggt gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt agcgctagct
     3481 tcgaaagatc tggatcccag aagcaaatgt aagcaataga tggctctgcc ctgactttta
     3541 tgcccagccc gagctccgaa aaagaagctt ccaaaagcag ctttcttttt gaaaattctg
     3601 tctttttata tagatttact gttttttttg attagatagg tgttttatat ataattgctc
     3661 aaataaccat atgcatggat accttgatat ttacataagg gattacctac agcaatgagg
     3721 ccagtttata tttgtttatg tgaggaaata cttggaagga ttatttaaat gccttgcaaa
     3781 tgttgccctc tggtggaagg tttaaaaaga atcttaaccc ttaattacat aactcaagct
     3841 actggttttt gtgtgtttat ttgttttttc tttctttccc tcttctttgt tagtatcagt
     3901 taaaataaaa tctataccta aaagcataaa gtccttcttg tgggcaatat gcatgcaatt
     3961 atcaaactga ttacaaaatt tctgttacag actcactgat ggcttgagaa tctcacttga
     4021 acacaaaaga aatttcacta tgctgaggtt tagaaaagcc agctaagcaa gcacacaggg
     4081 gtgttgtttc agatttgact ccacagtaga ttaatttacg attctcttgt agactagagc
     4141 ttcgattatt actcttacaa cgcgtgcggc cgcaggaacc cctagtgatg gagttggcca
     4201 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     4261 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggacatgtg agcaaaaggc
     4321 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4381 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4441 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4501 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4561 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4621 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4681 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4741 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4801 agaagaacag tatttggtat ctgcgctct
//
