LOCUS       sequence-425379-        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   142..271
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(333..921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(413..432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1092..1883,1884..1952))
                     /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     1715..1734
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1953..2057)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2125..2143
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2181..2203)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2303..2322
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2339..2794)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2476..2497)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2688..2707
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2869..3009
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2880..3009
                     /label=AAV2 ITR
     polyA_signal    3127..3238
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3425..3445
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3538..4248,4249..4251,4252..4254))
                     /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(3580..3601)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3929..3948
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4188..4209
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4251..4260
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 aaaaaaaaat ctcaccaagt atgtaagttt caaggctaaa tgtagactgt gccgagagcc
       61 agaaatgaac ataaaaggag ataccaacat ttcttattga gggaaatgag agatataatc
      121 ttagggctaa aatgcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
      181 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca
      241 gtgagcgagc gagcgcgcag ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc
      301 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
      361 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      421 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      481 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
      541 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
      601 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
      661 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
      721 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
      781 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
      841 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
      901 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
      961 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1021 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1081 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1141 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1201 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     1261 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     1321 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     1381 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     1441 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     1501 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     1561 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     1621 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     1681 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     1741 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     1801 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     1861 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     1921 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     1981 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2041 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     2101 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     2161 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     2221 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     2281 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataaa
     2341 attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     2401 tttaaccaat agaccgaaat cggcaaaatc ccttataaat caaaagaata gcccgagata
     2461 gagttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     2521 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcacccaaa
     2581 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     2641 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     2701 aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     2761 cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga cgtatgcggt
     2821 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgcccc tgcaggcagc
     2881 tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg
     2941 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
     3001 ggggttcctg cggccgctcg gtccgcacgt gagatctcat gcctgctatt gtcttcccaa
     3061 tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga
     3121 caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc accttccagg
     3181 gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagtc
     3241 gaagatctct cgagccacgg aattgtcagt gcccaacagc cgagcccctg tccagcagcg
     3301 ggcaaggcag gcggcgatga gttccgccgt ggcaagaact aaccaggatt tatacaagga
     3361 ggagaaaatg aaagccatac gggaagcaat agcatgatac aaaggcatta aagcagcgta
     3421 tccacatagc gtaaaaggag caacatagtt aagaatacca gtcaatcttt cacaaatttt
     3481 gtaatccaga ggttgattat gatatcgaat tcgcggccgc ggcgcgccgt cgacttactt
     3541 gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat
     3601 gtgatcgcgc ttctcgttgg ggtctttgct cagcttggac tggtagctca ggtagtggtt
     3661 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag
     3721 ctgcacgccg ccgtcctcga tgttgtggcg gatcttgaag ttggccttga tgccgttctt
     3781 ctgcttgtcg gcggtgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc
     3841 caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt
     3901 gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat
     3961 ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat
     4021 gtggtcgggg tagcgggcga agcactgcac gccgtagccc agggtggtca cgagggtggg
     4081 ccagggcacg ggcagcttgc cggtggtgca gatcagcttc agggtcagct tgccgtaggt
     4141 ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag
     4201 ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatggtggc
     4261 gaccggtagc gctagcttcg aaagatctgg atcccagaag caaatgtaag caatagatgg
     4321 ctctgccctg acttttatgc ccagcccgag ctccgaatgg tgcaatggag tttgtctata
     4381 ggttgcacag tccatgtcct caacctccat tcagactacc tccaagatac taggttcata
     4441 ctctttgata cgcaatgaat tataaatcac agaatctcat gttctctttc ttattgtgtc
     4501 agtattgatg tcacatgatt tctaacctgg aacatacact cagagttcac ctaattaact
     4561 cattctatgc tttactattg tcaaatatta caaatgagtt ccatttgcaa gataattttc
     4621 taaaaatttt aaaaccaaaa tggacagatt aatgatgaat atgtaagaga aagagagagg
     4681 tcaaggtatt gaatcccaga atgtctcgta caaagaaaaa gttatatcaa gaacagtagt
     4741 acagaataat gcaggtgcct cactacctga aaattatata acatgaact
//
