LOCUS       sequence-466249-        4915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2146m 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 4915)
  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 4915)
  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: AiP1375 - pAAV-AiE2146m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(671..1462,1463..1531))
                     /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     1294..1313
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1532..1636)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1704..1722
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1760..1782)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1882..1901
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1918..2373)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2055..2076)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2267..2286
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2448..2588
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2459..2588
                     /label=AAV2 ITR
     polyA_signal    2706..2817
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3004..3024
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3117..3827,3828..3830,3831..3833))
                     /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(3159..3180)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3508..3527
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3767..3788
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3830..3839
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4636..4765
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4827..4915,1..500))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(join(4907..4915,1..11))
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
       61 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
      121 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
      181 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
      241 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
      301 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
      361 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
      421 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
      481 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
      541 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
      601 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
      661 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
      721 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
      781 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
      841 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
      901 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
      961 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     1021 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     1081 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     1141 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     1201 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     1261 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     1321 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     1381 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     1441 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     1501 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     1561 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     1621 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     1681 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     1741 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     1801 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     1861 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccataaaa
     1921 ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     1981 ttaaccaata gaccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag
     2041 agttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     2101 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat
     2161 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     2221 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     2281 aaggagcggg cgctaaggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     2341 ccgccgcgct taatgcgccg ctacagggcg cgtactatgg ttgctttgac gtatgcggtg
     2401 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgcccct gcaggcagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttcctgc ggccgctcgg tccgcacgtg agatctcatg cctgctattg tcttcccaat
     2641 cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac
     2701 aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg
     2761 tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg
     2821 aagatctctc gagccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg
     2881 gcaaggcagg cggcgatgag ttccgccgtg gcaagaacta accaggattt atacaaggag
     2941 gagaaaatga aagccatacg ggaagcaata gcatgataca aaggcattaa agcagcgtat
     3001 ccacatagcg taaaaggagc aacatagtta agaataccag tcaatctttc acaaattttg
     3061 taatccagag gttgattatg atatcgaatt cgcggccgcg gcgcgccgtc gacttacttg
     3121 tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg
     3181 tgatcgcgct tctcgttggg gtctttgctc agcttggact ggtagctcag gtagtggttg
     3241 tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc
     3301 tgcacgccgc cgtcctcgat gttgtggcgg atcttgaagt tggccttgat gccgttcttc
     3361 tgcttgtcgg cggtgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc
     3421 aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg
     3481 tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg
     3541 gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg
     3601 tggtcggggt agcgggcgaa gcactgcacg ccgtagccca gggtggtcac gagggtgggc
     3661 cagggcacgg gcagcttgcc ggtggtgcag atcagcttca gggtcagctt gccgtaggtg
     3721 gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc
     3781 tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac catggtggcg
     3841 accggtagcg ctagcttcga aagatctgga tcccagaagc aaatgtaagc aatagatggc
     3901 tctgccctga cttttatgcc cagcccgagc tcggaagcaa ttgggagagc cagagcccag
     3961 atttgaaggg cagaggttag tgaactggaa ccactagtcc ccagggactc caggccttag
     4021 tgtgtgtctg gcctggtcct tgaccccatt agtccacagg agcccaagtg ccccaaaaca
     4081 accagctctg ttcagagagg aaagcacgtg acaagaagag agctgtcctc tactgaagag
     4141 ccccctagac tggaccagct gcagcagcaa gcagggtcct tttgccttca gccctctctg
     4201 tttatagcag tgtaacaggg tggggatggg ggatgggact ctgtgtcccg gcttatcagt
     4261 ttatggaaga atccttgcca cccactcctt ccaaggagac atctgagtat gcaccatacc
     4321 cccacactcc acccctactg gggcctgcta gtccaggaca ttccccttcc cccactgaag
     4381 cagtgcttcc tctccggcca gttctgttca cctccctaga ggctcttgat ttatgttgtt
     4441 gagtctgaaa agctaacctt ggagaaggaa aggaaataga gaaaaccagt cccagaggcc
     4501 agaggggctg gagccaggat cagagcccct tggaagagga tatttagtgc agcactagct
     4561 gaccacagca gcaccctggc tttggaaaag ctgaagctgc cttcaggaaa agcagctgag
     4621 tacgcgtgcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     4681 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc ctcagtgagc
     4741 gagcgagcgc gcagctgcct gcaggacatg tgagcaaaag gccagcaaaa ggccaggaac
     4801 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4861 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag atacc
//
