LOCUS       sequence-466268-        5130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2093m 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 5130)
  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 5130)
  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: AiP1508 - pAAV-AiE2093m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   427..556
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(618..1206)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(698..717)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1377..2168,2169..2237))
                     /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     2000..2019
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2238..2342)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2410..2428
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2466..2488)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2588..2607
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2624..3079)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2761..2782)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2973..2992
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3154..3294
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3165..3294
                     /label=AAV2 ITR
     polyA_signal    3412..3523
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3710..3730
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3823..4533,4534..4536,4537..4539))
                     /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(3865..3886)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4214..4233
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4473..4494
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4536..4545
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cacggtgaga atctgtggtt taatatacaa acaaggtgca tttgaagttg aagaagagat
       61 acagtctaca cccatgagga aacaaaacat cagaagaatc ctggcaggaa tgcacacggc
      121 tccatgttct gtcgaagaaa ccaatgccac agcaagttgc ttctgcagac gtgaagtgtg
      181 tactggaagc agcggtgctg ggctttgcat gggatttgca tggaggtaca cttcttacat
      241 ccatgacatc ttacatgtcg cttgcccagg tagatatctg catttctgag ctaagcatgc
      301 ataccatgat gtagttagag aaactgaggt cagagcagag tcaaggattg ttcttcctgc
      361 atgtctgagc actccaccca cccctagacc gccggcgatt attactctta caacgcgtgc
      421 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
      481 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg
      541 cgcagctgcc tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
      601 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
      661 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
      721 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
      781 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
      841 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
      901 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
      961 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     1021 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     1081 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     1141 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     1201 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     1261 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     1321 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     1381 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     1441 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     1501 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     1561 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     1621 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     1681 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     1741 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     1801 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     1861 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     1921 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     1981 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     2041 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     2101 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     2161 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     2221 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     2281 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     2341 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     2401 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     2461 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     2521 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     2581 aactatgcgg catcagagca gattgtactg agagtgcacc ataaaattgt aaacgttaat
     2641 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaatagacc
     2701 gaaatcggca aaatccctta taaatcaaaa gaatagcccg agatagagtt gagtgttgtt
     2761 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2821 accgtctatc agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg
     2881 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2941 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     3001 aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3061 gcgccgctac agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca
     3121 gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc
     3181 tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag
     3241 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc
     3301 gctcggtccg cacgtgagat ctcatgcctg ctattgtctt cccaatcctc ccccttgctg
     3361 tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt
     3421 tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg
     3481 ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaaga tctctcgagc
     3541 cacggaattg tcagtgccca acagccgagc ccctgtccag cagcgggcaa ggcaggcggc
     3601 gatgagttcc gccgtggcaa gaactaacca ggatttatac aaggaggaga aaatgaaagc
     3661 catacgggaa gcaatagcat gatacaaagg cattaaagca gcgtatccac atagcgtaaa
     3721 aggagcaaca tagttaagaa taccagtcaa tctttcacaa attttgtaat ccagaggttg
     3781 attatgatat cgaattcgcg gccgcggcgc gccgtcgact tacttgtaca gctcgtccat
     3841 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
     3901 gttggggtct ttgctcagct tggactggta gctcaggtag tggttgtcgg gcagcagcac
     3961 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgccgccgtc
     4021 ctcgatgttg tggcggatct tgaagttggc cttgatgccg ttcttctgct tgtcggcggt
     4081 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
     4141 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     4201 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
     4261 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     4321 ggcgaagcac tgcacgccgt agcccagggt ggtcacgagg gtgggccagg gcacgggcag
     4381 cttgccggtg gtgcagatca gcttcagggt cagcttgccg taggtggcat cgccctcgcc
     4441 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     4501 caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg gtagcgctag
     4561 cttcgaaaga tctggatccc agaagcaaat gtaagcaata gatggctctg ccctgacttt
     4621 tatgcccagc ccgagctccg ccggcgccaa acatatccct ggcttctttt tgttttttat
     4681 aaaacacaac agtaggtact tcaccaacta agctatctcc ctagcctagt agccatttga
     4741 aatatcagga tacactgctc ttcttcaaga aatcattggg tgacacagcc ataccaaacc
     4801 catatcctca gaggtccaaa ctcaagtctt gccaaatgcc aactagcagc tggctacagg
     4861 aaggacaggc tcctgtctct aatttttcca atgtaagagc tcaccatgtg tcacagtccc
     4921 taccgccctc tgcctgtcca cactggggct aagtgtcaac agccaggtgt tatcacatcc
     4981 agagtggggc gttttctatc ctataataaa aaagaaagaa gcatttatct ttatacccat
     5041 gtgtttttca gaagtggaaa aattcccaaa gactgagcca gccgcccggc tttgaaagaa
     5101 acgcatgtgc tagactgatt ttttaaagaa
//
