LOCUS       sequence-432589-        4789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2588m 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 4789)
  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 4789)
  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: AiP1959 - pAAV-AiE2588m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   77..217
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   88..217
                     /label=AAV2 ITR
     polyA_signal    335..446
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     633..653
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(746..1456,1457..1459,1460..1462))
                     /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(788..809)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1137..1156
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1396..1417
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1459..1468
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2139..2268
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2330..2918)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2410..2429)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3089..3880,3881..3949))
                     /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     3712..3731
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3950..4054)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4122..4140
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4178..4200)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4300..4319
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4336..4789,1..2))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4473..4494)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4685..4704
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtactatggt tgctttgacg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
       61 taccgcatca ggcgcccctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg
      121 gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc
      181 agagagggag tggccaactc catcactagg ggttcctgcg gccgctcggt ccgcacgtga
      241 gatctcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc
      301 ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga
      361 aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac
      421 agatggctgg caactagaag gcacagtcga agatctctcg agccacggaa ttgtcagtgc
      481 ccaacagccg agcccctgtc cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg
      541 caagaactaa ccaggattta tacaaggagg agaaaatgaa agccatacgg gaagcaatag
      601 catgatacaa aggcattaaa gcagcgtatc cacatagcgt aaaaggagca acatagttaa
      661 gaataccagt caatctttca caaattttgt aatccagagg ttgattatga tatcgaattc
      721 gcggccgcgg cgcgccgtcg acttacttgt acagctcgtc catgccgaga gtgatcccgg
      781 cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca
      841 gcttggactg gtagctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg
      901 gggtgttctg ctggtagtgg tcggcgagct gcacgccgcc gtcctcgatg ttgtggcgga
      961 tcttgaagtt ggccttgatg ccgttcttct gcttgtcggc ggtgatatag acgttgtggc
     1021 tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
     1081 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
     1141 tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg
     1201 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcgggcgaag cactgcacgc
     1261 cgtagcccag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
     1321 tcagcttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga
     1381 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
     1441 gctcctcgcc cttgctcacc atggtggcga ccggtagcgc tagcttcgaa agatctggat
     1501 cccagaagca aatgtaagca atagatggct ctgccctgac ttttatgccc agcccgagct
     1561 ccgaatggtg caatggagtt tgtctatagg ttgcacagtc catgtcctca acctccattc
     1621 agactacctc caagatacta ggttcatact ctttgatacg caatgaatta taaatcacag
     1681 aatctcatgt tctctttctt attgtgtcag tattgatgtc acatgatttc taacctggaa
     1741 catacactca gagttcacct aattaactca ttctatgctt tactattgtc aaatattaca
     1801 aatgagttcc atttgcaaga taattttcta aaaattttaa aaccaaaatg gacagattaa
     1861 tgatgaatat gtaagagaaa gagagaggtc aaggtattga atcccagaat gtctcgtaca
     1921 aagaaaaagt tatatcaaga acagtagtac agaataatgc aggtgcctca ctacctgaaa
     1981 attatataac atgaactaaa aaaaaatctc accaagtatg taagtttcaa ggctaaatgt
     2041 agactgtgcc gagagccaga aatgaacata aaaggagata ccaacatttc ttattgaggg
     2101 aaatgagaga tataatctta gggctaaaat gcggccgcag gaacccctag tgatggagtt
     2161 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     2221 acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcaggac atgtgagcaa
     2281 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2341 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2401 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2461 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2521 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2581 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2641 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2701 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2761 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2821 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2881 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2941 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3001 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3061 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3121 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3181 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3241 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3301 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3361 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3421 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3481 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3541 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3601 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3661 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3721 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3781 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3841 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3901 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3961 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4021 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4081 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     4141 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     4201 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     4261 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     4321 tgagagtgca ccataaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt
     4381 gttaaatcag ctcatttttt aaccaataga ccgaaatcgg caaaatccct tataaatcaa
     4441 aagaatagcc cgagatagag ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4501 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4561 gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4621 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4681 aggaagggaa gaaagcgaaa ggagcgggcg ctaaggcgct ggcaagtgta gcggtcacgc
     4741 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgc
//
