LOCUS       sequence-432592-        4749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2447m 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 4749)
  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 4749)
  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: AiP1921 - pAAV-AiE2447m-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   179..308
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(370..958)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(450..469)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1129..1920,1921..1989))
                     /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     1752..1771
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1990..2094)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2162..2180
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2218..2240)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2340..2359
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2376..2831)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2513..2534)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2725..2744
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2906..3046
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2917..3046
                     /label=AAV2 ITR
     polyA_signal    3164..3275
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3462..3482
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3575..4285,4286..4288,4289..4291))
                     /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(3617..3638)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3966..3985
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4225..4246
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4288..4297
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 agtcccctcc agggcacagg ccatgtcccc ccagcccatg taacaaagta tactatctac
       61 tcactgcccc agtgtatgtg ctggcactgg ggctgggtcc tcaggcaggt cctatgagac
      121 acagaatgtt tcagaagatt attcctccga ttattactct tacaacgcgt gcggccgcag
      181 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
      241 gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
      301 cctgcaggac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      361 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      421 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      481 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      541 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      601 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
      661 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      721 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      781 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
      841 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
      901 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
      961 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1021 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1081 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1141 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1201 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1261 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1321 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     1381 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     1441 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     1501 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     1561 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     1621 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     1681 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     1741 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     1801 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     1861 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     1921 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     1981 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2041 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     2101 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     2161 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     2221 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     2281 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     2341 ggcatcagag cagattgtac tgagagtgca ccataaaatt gtaaacgtta atattttgtt
     2401 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaataga ccgaaatcgg
     2461 caaaatccct tataaatcaa aagaatagcc cgagatagag ttgagtgttg ttccagtttg
     2521 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2581 tcagggcgat ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg
     2641 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2701 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctaaggcgct
     2761 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     2821 acagggcgcg tactatggtt gctttgacgt atgcggtgtg aaataccgca cagatgcgta
     2881 aggagaaaat accgcatcag gcgcccctgc aggcagctgc gcgctcgctc gctcactgag
     2941 gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
     3001 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgctcggtc
     3061 cgcacgtgag atctcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc
     3121 caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt
     3181 ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg
     3241 gcaaacaaca gatggctggc aactagaagg cacagtcgaa gatctctcga gccacggaat
     3301 tgtcagtgcc caacagccga gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt
     3361 ccgccgtggc aagaactaac caggatttat acaaggagga gaaaatgaaa gccatacggg
     3421 aagcaatagc atgatacaaa ggcattaaag cagcgtatcc acatagcgta aaaggagcaa
     3481 catagttaag aataccagtc aatctttcac aaattttgta atccagaggt tgattatgat
     3541 atcgaattcg cggccgcggc gcgccgtcga cttacttgta cagctcgtcc atgccgagag
     3601 tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt
     3661 ctttgctcag cttggactgg tagctcaggt agtggttgtc gggcagcagc acggggccgt
     3721 cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgccgccg tcctcgatgt
     3781 tgtggcggat cttgaagttg gccttgatgc cgttcttctg cttgtcggcg gtgatataga
     3841 cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga
     3901 agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg
     3961 cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt
     4021 cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cgggcgaagc
     4081 actgcacgcc gtagcccagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg
     4141 tggtgcagat cagcttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg
     4201 acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc
     4261 cggtgaacag ctcctcgccc ttgctcacca tggtggcgac cggtagcggc aagctggtgg
     4321 tcccgagttc tgtctccaca gcccttgcca ggcaggcggc ctcttaaagg ccaggctgac
     4381 gtcaaagccc ctctgggtcc cccacctcct acctcctgcg catcctctcc acgcctgctg
     4441 tgcctgaggg aggcgggggt gggcacagag cgagggcgac ggacgagaga ggcctccgtc
     4501 ccattagatc taattgcatc cactgtcgca ggcacctgcc tctgaatcac cctccgccct
     4561 agacacgaca tgaagacagg ggctggctgg tgttctggga caggagacat ctagagctct
     4621 ctaatcaaac cagggcatat gggagcgccc atgctggatt acccaagggt tcatgttagg
     4681 aaggctatat atagcccttg gagacaaata aatctaggag ccctctcact agctttttgg
     4741 atgtcttca
//
