LOCUS       sequence-466319-        5040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2423m 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 5040)
  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 5040)
  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: AiP1877 - pAAV-AiE2423m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     77..95
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(133..155)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     255..274
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(291..746)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(428..449)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     640..659
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   821..961
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   832..961
                     /label=AAV2 ITR
     polyA_signal    1079..1190
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1377..1397
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1490..2200,2201..2203,2204..2206))
                     /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(1532..1553)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1881..1900
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2140..2161
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2203..2212
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3134..3263
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3325..3913)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3405..3424)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4084..4875,4876..4944))
                     /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     4707..4726
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4945..5040,1..9))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
       61 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
      121 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
      181 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
      241 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata aaattgtaaa
      301 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
      361 atagaccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga tagagttgag
      421 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
      481 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt
      541 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
      601 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
      661 gggcgctaag gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
      721 gcttaatgcg ccgctacagg gcgcgtacta tggttgcttt gacgtatgcg gtgtgaaata
      781 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc cctgcaggca gctgcgcgct
      841 cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg
      901 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
      961 tgcggccgct cggtccgcac gtgagatctc atgcctgcta ttgtcttccc aatcctcccc
     1021 cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga
     1081 tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga
     1141 aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaagatct
     1201 ctcgagccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc
     1261 aggcggcgat gagttccgcc gtggcaagaa ctaaccagga tttatacaag gaggagaaaa
     1321 tgaaagccat acgggaagca atagcatgat acaaaggcat taaagcagcg tatccacata
     1381 gcgtaaaagg agcaacatag ttaagaatac cagtcaatct ttcacaaatt ttgtaatcca
     1441 gaggttgatt atgatatcga attcgcggcc gcggcgcgcc gtcgacttac ttgtacagct
     1501 cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc
     1561 gcttctcgtt ggggtctttg ctcagcttgg actggtagct caggtagtgg ttgtcgggca
     1621 gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc
     1681 cgccgtcctc gatgttgtgg cggatcttga agttggcctt gatgccgttc ttctgcttgt
     1741 cggcggtgat atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt
     1801 tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct
     1861 cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct
     1921 cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg
     1981 ggtagcgggc gaagcactgc acgccgtagc ccagggtggt cacgagggtg ggccagggca
     2041 cgggcagctt gccggtggtg cagatcagct tcagggtcag cttgccgtag gtggcatcgc
     2101 cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca
     2161 ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggtg gcgaccggta
     2221 gcgctagctt cgaaagatct ggatcccaga agcaaatgta agcaatagat ggctctgccc
     2281 tgacttttat gcccagcccg agctccggta ctgttttgta tccattttgt accctgtcag
     2341 acttgacact ctgcctgctc attcattcat tcaaactagt tttagaggca gaccccgtat
     2401 agattggaat gatgctagaa agactttgtg agcacccaag agctcaggag ctcgagcctg
     2461 gagagagcat tctttctttt tttttctaga aaatcatttg atctcttgac ctgttaatat
     2521 tttgggagac ttctcagccc cgtcttgcta cccaccccct tccgattaaa actgcacact
     2581 tttctgtctg gagcaactct ggctaatctg gagagggaag acaaagttta gatctggttg
     2641 agatttggtt acatttctaa aagagaacct ccaaaagctt ccagatttgc aggcgtaaga
     2701 taaagacagc cctgacattt gccgggaggt acggtgatcg ctgcttctca gcagttcatt
     2761 ttttttcccc caggcactgc tggcttcctt tgactattat agttgccaga aaaatccttg
     2821 ctttttttac tttacaacca gcatttgagt ggcaagttgg atataatggg atgagaccaa
     2881 atctctccat tcctcctggg agcaatgatg ggacacaatt tccagtccca caagttctga
     2941 atcttgtagc gtgtagcact gtcaggcctg ttcttgtcgg ggtgggcagt agtgacggta
     3001 tttgcaaaga gataataaaa aagaactggg cagacttccg ttaactttaa ttatgaaatt
     3061 gaaataaggc atgccacttt aatattggag gctggtctgt ttcgattatt actcttacaa
     3121 cgcgtgcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3181 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcggcct cagtgagcga
     3241 gcgagcgcgc agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3301 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3361 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3421 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3481 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3541 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3601 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3661 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3721 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3781 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3841 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3901 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3961 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4021 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4081 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4141 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4201 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4261 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4321 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4381 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4441 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4501 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4561 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4621 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4681 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4741 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4801 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4861 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4921 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4981 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
//
