LOCUS       sequence-432573-        4663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2564m 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 4663)
  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 4663)
  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: AiP1860 - pAAV-AiE2564m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     101..119
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(157..179)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     279..298
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(315..770)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(452..473)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     664..683
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   845..985
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   856..985
                     /label=AAV2 ITR
     polyA_signal    1103..1214
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1401..1421
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1514..2224,2225..2227,2228..2230))
                     /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(1556..1577)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1905..1924
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2164..2185
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2227..2236
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2781..2910
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2972..3560)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3052..3071)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3731..4522,4523..4591))
                     /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     4354..4373
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4592..4663,1..33))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
       61 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
      121 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
      181 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
      241 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
      301 gagagtgcac cataaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg
      361 ttaaatcagc tcatttttta accaatagac cgaaatcggc aaaatccctt ataaatcaaa
      421 agaatagccc gagatagagt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
      481 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
      541 tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
      601 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
      661 ggaagggaag aaagcgaaag gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct
      721 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg
      781 ctttgacgta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
      841 cgcccctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
      901 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      961 gccaactcca tcactagggg ttcctgcggc cgctcggtcc gcacgtgaga tctcatgcct
     1021 gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga
     1081 atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg
     1141 agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca
     1201 actagaaggc acagtcgaag atctctcgag ccacggaatt gtcagtgccc aacagccgag
     1261 cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca agaactaacc
     1321 aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag
     1381 gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca
     1441 atctttcaca aattttgtaa tccagaggtt gattatgata tcgaattcgc ggccgcggcg
     1501 cgccgtcgac ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
     1561 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggt
     1621 agctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
     1681 ggtagtggtc ggcgagctgc acgccgccgt cctcgatgtt gtggcggatc ttgaagttgg
     1741 ccttgatgcc gttcttctgc ttgtcggcgg tgatatagac gttgtggctg ttgtagttgt
     1801 actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga
     1861 tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt
     1921 cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga
     1981 agtcgtgctg cttcatgtgg tcggggtagc gggcgaagca ctgcacgccg tagcccaggg
     2041 tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatc agcttcaggg
     2101 tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt
     2161 ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct
     2221 tgctcaccat ggtggcgacc ggtagcgcta gcttcgaaag atctggatcc cagaagcaaa
     2281 tgtaagcaat agatggctct gccctgactt ttatgcccag cccgagctcc gtggggatag
     2341 gtttcttagg agtctccaag gatcaggatg ggatgttgag gcacagggag aaatcaatgg
     2401 tggctcttct attttagtct ctagttaaat ggtgccagct atggagatgg tgagatgggg
     2461 cggctacact ggagaggaaa gccggaagtc tattttgaat gtattatggt tccaagaggg
     2521 attgtgtgaa ggaagcagtg ggatagagat tagagctgct caggatgtgg agggggaggc
     2581 tgagagaaga tcaggactta gaatggaact ggtacttaaa ggcccctgga agcaagatag
     2641 gaaaccgaac tggattctga tgacgccatg ctcagtttaa ctggcgaccg gaaacctcct
     2701 tcctttttga aagacattgc tgctgttact ctttccattt ggattctccc gattattact
     2761 cttacaacgc gtgcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     2821 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag
     2881 tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc aaaaggccag caaaaggcca
     2941 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3001 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3061 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3121 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3181 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3241 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3301 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3361 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3421 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3481 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3541 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3601 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3661 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3721 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3781 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3841 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3901 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3961 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4021 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4081 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4141 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4201 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4261 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4321 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4381 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4441 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4501 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4561 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4621 tttatcaggg ttattgtctc atgagcggat acatatttga atg
//
