LOCUS       sequence-466310-        5238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2438m 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 5238)
  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 5238)
  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: AiP1842 - pAAV-AiE2438m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    139..250
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     437..457
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(550..1260,1261..1263,1264..1266))
                     /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(592..613)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     941..960
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1200..1221
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1263..1272
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2392..2521
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2583..3171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2663..2682)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3342..4133,4134..4202))
                     /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     3965..3984
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4203..4307)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4375..4393
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4431..4453)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4553..4572
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4589..5044)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4726..4747)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4938..4957
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   join(5119..5238,1..21)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(5130..5238,1..21)
                     /label=AAV2 ITR
ORIGIN
        1 actccatcac taggggttcc tgcggccgct cggtccgcac gtgagatctc atgcctgcta
       61 ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc caccccccag aatagaatga
      121 cacctactca gacaatgcga tgcaatttcc tcattttatt aggaaaggac agtgggagtg
      181 gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta
      241 gaaggcacag tcgaagatct ctcgagccac ggaattgtca gtgcccaaca gccgagcccc
      301 tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaagaa ctaaccagga
      361 tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat acaaaggcat
      421 taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac cagtcaatct
      481 ttcacaaatt ttgtaatcca gaggttgatt atgatatcga attcgcggcc gcggcgcgcc
      541 gtcgacttac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc
      601 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg actggtagct
      661 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta
      721 gtggtcggcg agctgcacgc cgccgtcctc gatgttgtgg cggatcttga agttggcctt
      781 gatgccgttc ttctgcttgt cggcggtgat atagacgttg tggctgttgt agttgtactc
      841 cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg
      901 gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc
      961 cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc
     1021 gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc acgccgtagc ccagggtggt
     1081 cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatcagct tcagggtcag
     1141 cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac
     1201 gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct
     1261 caccatggtg gcgaccggta gcgctagctt cgaaagatct ggatcccaga agcaaatgta
     1321 agcaatagat ggctctgccc tgacttttat gcccagcccg agctccgtgt gtgtgtgtgt
     1381 acaggcaagg caggcacagg tgggatcccc cagccccttg tgaaaagccc aagtgagaat
     1441 ttgtcaggta ggtcacagtc tggtggcagc ccctgcaaca tctcatttcc aaaatgtcca
     1501 ctgccaggcc ctagctgtgg tcaggggagt gttcatacaa caggacaagc ctcgggttgt
     1561 tgctgagcag aggttaatat ctctcaggag caaggacaag catgcgtgaa agctctgtac
     1621 ctccagcgat ggtgggcagc agggtctagg atacactggg tgtatctctg cttcgacctt
     1681 gtgctcattt tcagggagat ctatttctgg caagcaatat tggtctttta gttacaatat
     1741 gatgtaaagt ttccttttga gatactctta tttaatctaa aaaaaggaaa tatttaagta
     1801 aataaaaata gacatggcac acagaagttt gcccagctct tgtggctggc ttgcaaatga
     1861 cttcagtttc tgtggcctaa tttagaaaaa tgcttcctct atgtattcat tgactcaaca
     1921 aatatttatt gaacacctac tgggtgtgag acactggggg aggactaaac gaagacacca
     1981 gagtcgggag attccattca ggaatcaagg gcctgagtgg acaactcacc cacacaccac
     2041 ataaagaaag aaaaccgaga cttgatcctg agagcactgg ggagctgctg ggacgtaaga
     2101 tcaagtgtgt ggaggctaag gctggcccat gtacaggatg aagtaaaggg agagaagact
     2161 ggaagccaac agcccggggg atgggggtgg ggtacagatt tgtgataggt gtaagggctg
     2221 gaaggactgg tacctagctg agttcctagc acaaagtagg cactcaatac atggctggaa
     2281 ctattgttgt tactattatt aaaatctcct ctcttcctta acttggagaa ttaattaagt
     2341 cttagttcca tgaatcatca cgattattac tcttacaacg cgtgcggccg caggaacccc
     2401 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     2461 caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     2521 gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2581 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2641 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2701 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2761 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2821 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2881 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2941 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3001 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3061 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3121 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3181 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3241 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3301 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3361 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3421 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3481 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3541 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3601 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3661 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3721 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3781 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3841 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3901 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3961 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4021 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4081 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4141 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4201 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4261 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4321 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     4381 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     4441 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     4501 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     4561 gagcagattg tactgagagt gcaccataaa attgtaaacg ttaatatttt gttaaaattc
     4621 gcgttaaatt tttgttaaat cagctcattt tttaaccaat agaccgaaat cggcaaaatc
     4681 ccttataaat caaaagaata gcccgagata gagttgagtg ttgttccagt ttggaacaag
     4741 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     4801 gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa
     4861 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     4921 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt
     4981 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     5041 gcgtactatg gttgctttga cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     5101 aataccgcat caggcgcccc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc
     5161 gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
     5221 gcagagaggg agtggcca
//
