LOCUS       sequence-470452-        4747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0487m 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 4747)
  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 4747)
  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: AiP12262 - pAAV-AiE0487m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(242..1033,1034..1102))
                     /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     865..884
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1103..1207)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1275..1293
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1331..1353)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1453..1472
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1489..1944)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1626..1647)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1838..1857
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2019..2159
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2030..2159
                     /label=AAV2 ITR
     polyA_signal    2277..2388
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2575..2595
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2688..3398,3399..3401,3402..3404))
                     /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(2730..2751)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3079..3098
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3338..3359
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3401..3410
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4039..4168
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4230..4747,1..71))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4310..4329)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
       61 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
      121 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
      181 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
      241 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
      301 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
      361 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
      421 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
      481 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
      541 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
      601 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
      661 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
      721 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
      781 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
      841 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
      901 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
      961 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     1021 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     1081 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     1141 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     1201 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     1261 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     1321 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     1381 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     1441 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataaa attgtaaacg
     1501 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     1561 agaccgaaat cggcaaaatc ccttataaat caaaagaata gcccgagata gagttgagtg
     1621 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     1681 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcacccaaa tcaagttttt
     1741 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     1801 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     1861 gcgctaaggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     1921 ttaatgcgcc gctacagggc gcgtactatg gttgctttga cgtatgcggt gtgaaatacc
     1981 gcacagatgc gtaaggagaa aataccgcat caggcgcccc tgcaggcagc tgcgcgctcg
     2041 ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc
     2101 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
     2161 cggccgctcg gtccgcacgt gagatctcat gcctgctatt gtcttcccaa tcctccccct
     2221 tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg
     2281 caatttcctc attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag
     2341 gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagtc gaagatctct
     2401 cgagccacgg aattgtcagt gcccaacagc cgagcccctg tccagcagcg ggcaaggcag
     2461 gcggcgatga gttccgccgt ggcaagaact aaccaggatt tatacaagga ggagaaaatg
     2521 aaagccatac gggaagcaat agcatgatac aaaggcatta aagcagcgta tccacatagc
     2581 gtaaaaggag caacatagtt aagaatacca gtcaatcttt cacaaatttt gtaatccaga
     2641 ggttgattat gatatcgaat tcgcggccgc ggcgcgccgt cgacttactt gtacagctcg
     2701 tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc
     2761 ttctcgttgg ggtctttgct cagcttggac tggtagctca ggtagtggtt gtcgggcagc
     2821 agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgccg
     2881 ccgtcctcga tgttgtggcg gatcttgaag ttggccttga tgccgttctt ctgcttgtcg
     2941 gcggtgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg
     3001 ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg
     3061 aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc
     3121 tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg
     3181 tagcgggcga agcactgcac gccgtagccc agggtggtca cgagggtggg ccagggcacg
     3241 ggcagcttgc cggtggtgca gatcagcttc agggtcagct tgccgtaggt ggcatcgccc
     3301 tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg
     3361 atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatggtggc gaccggtagc
     3421 gctagcttcg aaagatctgg atcccagaag caaatgtaag caatagatgg ctctgccctg
     3481 acttttatgc ccagcccgag ctcgggcact tccttgttgg tgtagagaaa ggtccaaaga
     3541 gtctcgacta gtttctagga actgcaaacg caaaaggtca ttttggggga agaacattgt
     3601 tttgtttggt taggtttgat ttgggattta ctccacacag gttccagtgt ggagaagata
     3661 atggttgtca ctctctgact ccagggagat ttatgggcag gctctgtgtt acacatttcc
     3721 tcctgttgct ttttctcttt atgcccagcc ttttccagtt tattcagctg ctctcataat
     3781 tatccccaat tatttctaat ggtaggtttc tgaagctgga gttagagaaa cgccatgctt
     3841 tttgagcctt aaacatcact tagtggaaat ggactccaac aactcgcatg aagcttgagc
     3901 ccaggtcttc tcggggaatt ttttttcttt agatctgctt tcatcccgac tgtcttattt
     3961 ttctgacact ttacagaggt tgtactatta aggcagagac ggacgtggtg atgcacgcct
     4021 gtaaacgcgt gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     4081 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg
     4141 agcgagcgag cgcgcagctg cctgcaggac atgtgagcaa aaggccagca aaaggccagg
     4201 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4261 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4321 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4381 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4441 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4501 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4561 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4621 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4681 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4741 ggcaaac
//
