LOCUS       sequence-438946-        4758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2464m 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 4758)
  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 4758)
  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: AiP1882 - pAAV-AiE2464m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     190..209
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   371..511
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   382..511
                     /label=AAV2 ITR
     polyA_signal    629..740
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     927..947
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1040..1750,1751..1753,1754..1756))
                     /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(1082..1103)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1431..1450
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1690..1711
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1753..1762
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2402..2531
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2593..3181)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2673..2692)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3352..4143,4144..4212))
                     /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     3975..3994
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4213..4317)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4385..4403
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4441..4463)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4563..4582
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4599..4758,1..296))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4736..4757)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
       61 actacgtgaa ccatcaccca aatcaagttt tttggggtcg aggtgccgta aagcactaaa
      121 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
      181 gagaaaggaa gggaagaaag cgaaaggagc gggcgctaag gcgctggcaa gtgtagcggt
      241 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtacta
      301 tggttgcttt gacgtatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
      361 atcaggcgcc cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag
      421 cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag
      481 ggagtggcca actccatcac taggggttcc tgcggccgct cggtccgcac gtgagatctc
      541 atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc caccccccag
      601 aatagaatga cacctactca gacaatgcga tgcaatttcc tcattttatt aggaaaggac
      661 agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg
      721 ctggcaacta gaaggcacag tcgaagatct ctcgagccac ggaattgtca gtgcccaaca
      781 gccgagcccc tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaagaa
      841 ctaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat
      901 acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac
      961 cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt atgatatcga attcgcggcc
     1021 gcggcgcgcc gtcgacttac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
     1081 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg
     1141 actggtagct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
     1201 tctgctggta gtggtcggcg agctgcacgc cgccgtcctc gatgttgtgg cggatcttga
     1261 agttggcctt gatgccgttc ttctgcttgt cggcggtgat atagacgttg tggctgttgt
     1321 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
     1381 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     1441 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
     1501 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc acgccgtagc
     1561 ccagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatcagct
     1621 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
     1681 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     1741 cgcccttgct caccatggtg gcgaccggta gcgctagctt cgaaagatct ggatcccaga
     1801 agcaaatgta agcaatagat ggctctgccc tgacttttat gcccagcccg agctccgaat
     1861 tgcactttgt gacccccctc tttctcatgt aagctgagaa aggcaagaac acttgtctct
     1921 gcctccatcc tgctcattag actctttcct tttagtttgt ccaacacata ccattctagc
     1981 ataacttgtt ttagaaaatt attttatgtg gagtgctatg cttttacaga taacattatc
     2041 atgttaactc tgaagagtga gcaatatgga gattccaggt gcgttggaag aaacagaaga
     2101 aaacatttca aacttttcaa gccacaatag tctgctgcct cctcagctta tgtcaaaggc
     2161 tggctttgcc aactcctgcc tacatggggc tacattaaga ataactctcc cttctttgta
     2221 gccatagcta aaatactcca tgaccacaat tttgaagaat actagttggt tttctgtcat
     2281 tttatttgtg aaaataatca agataaaatg caacagcatc attttagaga tctgggttca
     2341 aggtggacta gtatgtactt gtataattat tttttttttc tgagttactg tttgcggccg
     2401 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     2461 gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag
     2521 ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2581 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2641 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2701 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2761 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2821 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2881 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2941 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3001 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3061 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3121 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3181 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3241 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3301 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3361 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3421 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3481 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3541 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3601 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3661 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3721 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3781 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3841 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3901 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3961 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4021 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4081 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4141 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4201 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4261 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4321 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     4381 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     4441 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     4501 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     4561 tgcggcatca gagcagattg tactgagagt gcaccataaa attgtaaacg ttaatatttt
     4621 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat agaccgaaat
     4681 cggcaaaatc ccttataaat caaaagaata gcccgagata gagttgagtg ttgttccagt
     4741 ttggaacaag agtccact
//
