LOCUS       sequence-432544-        4892 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2346m 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 4892)
  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 4892)
  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: AiP1663 - pAAV-AiE2346m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     49..68
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   210..339
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1113..1122
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1119..1121,1122..1124,1125..1835)
                     /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(1164..1185)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1425..1444)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1772..1793
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1928..1948)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2135..2246
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2364..2504
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2364..2493
                     /label=AAV2 ITR
     rep_origin      2579..3034
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2666..2685)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2876..2897
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3051..3070)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3170..3192
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3230..3248)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3316..3420
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3421..3489,3490..4281)
                     /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     complement(3639..3658)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4452..4892,1..148)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
       61 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
      121 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      181 ggccttttgc tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac
      241 tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
      301 gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgcacg cgttgtaaga
      361 gtaataatcg tatggaaccc actttggtct ccaactcaca tttcccctgc tttagtctcc
      421 agagtatggt gtctttgtta tgatcttgag ggatgtgaaa cactggcact ctccaaatcc
      481 accggtgcta tgtagggaga agcaggaggc ctaagggaga gagctggccg acatggacac
      541 ggagtggaag gcaggaagga aaagcctcta gcctctcacc ccgtgaggga ctccactcct
      601 taaagctgtc ccccaggtct aaaacttcct gtgttaggaa aggtgcttta gaaagaacat
      661 cccatctaag agcagctaag agcaagaaca tttattcgag tgtgaacaat ccagaagtca
      721 gcgattgcag ttacgctggc tccgagtcag ccgtctcctc ccctccggat cacaagctgg
      781 ctgtcagggt tccaagcttc atgttttcac ccagcagcgg cactagacag aaagggaaag
      841 gaggccaaga gagctaccag cacatctctt cgagggagca aagtggggcc ctgaagctac
      901 ccttgtaagt cccacgacat ggaaatgaac tcacaagtct caaaactgta ccggctacgc
      961 ccttttctta tctgtaagac tgactacgcc ccacgtgatg gactaagctt tagggcccgg
     1021 agctcgggct gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg
     1081 atccagatct ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc
     1141 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1201 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga
     1261 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg
     1321 gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1381 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1441 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1501 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1561 gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga
     1621 tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc
     1681 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc
     1741 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1801 ccgggatcac tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc
     1861 gaattcgata tcataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     1921 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     1981 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg
     2041 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2101 gcactgacaa ttccgtggct cgagagatct tcgactgtgc cttctagttg ccagccatct
     2161 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2221 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2281 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc
     2341 tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     2401 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     2461 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     2521 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     2581 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2641 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2701 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2761 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     2821 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2881 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     2941 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3001 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     3061 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     3121 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3181 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     3241 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     3301 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3361 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3421 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3481 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3541 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3601 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3661 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3721 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3781 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3841 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3901 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     3961 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4021 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4081 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4141 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4201 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4261 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4321 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4381 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4441 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4501 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4561 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4621 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4681 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4741 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4801 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4861 cttcccgaag ggagaaaggc ggacaggtat cc
//
