LOCUS       sequence-466270-        5110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2194m 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 5110)
  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 5110)
  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: AiP1519 - pAAV-AiE2194m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   417..546
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(608..1196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(688..707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1367..2158,2159..2227))
                     /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     1990..2009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2228..2332)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2400..2418
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2456..2478)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2578..2597
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2614..3069)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2751..2772)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2963..2982
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3144..3284
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3155..3284
                     /label=AAV2 ITR
     polyA_signal    3402..3513
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3700..3720
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3813..4523,4524..4526,4527..4529))
                     /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(3855..3876)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4204..4223
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4463..4484
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4526..4535
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctctgagcca agttctagca tctgctcagg aaagataatg ccagagcagg tatgttctgg
       61 aggaagctgt gagcactgag atccagcgtg agacttagac ccaacctggg gacaggaccc
      121 ggcacaagac agacttcagg taaatgtcaa atgccaaaag aagacagagg acacaagctg
      181 agatatatat tcaaaattca gagcacagtg caagggcaat taggtcaagc aggaaaaaaa
      241 gagaaagaga aggagaaagg agaaatcagt acagaaacta ccctctcttt atagtgggta
      301 tcattcgcca ggcattggca cccaatgccg agcataacta acgaggcaat aaggatggag
      361 atttaccaga aatatgaatc gccggcgatt attactctta caacgcgtgc ggccgcagga
      421 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
      481 gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
      541 tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      601 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      661 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      721 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      781 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      841 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
      901 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
      961 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1021 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1081 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1141 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1201 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1261 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     1321 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     1381 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     1441 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     1501 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     1561 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     1621 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     1681 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     1741 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     1801 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     1861 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     1921 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     1981 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     2041 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     2101 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     2161 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     2221 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     2281 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     2341 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     2401 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     2461 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     2521 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     2581 catcagagca gattgtactg agagtgcacc ataaaattgt aaacgttaat attttgttaa
     2641 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaatagacc gaaatcggca
     2701 aaatccctta taaatcaaaa gaatagcccg agatagagtt gagtgttgtt ccagtttgga
     2761 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     2821 agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc
     2881 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     2941 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg
     3001 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3061 agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag
     3121 gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc
     3181 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
     3241 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc gctcggtccg
     3301 cacgtgagat ctcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca
     3361 ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt
     3421 attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc
     3481 aaacaacaga tggctggcaa ctagaaggca cagtcgaaga tctctcgagc cacggaattg
     3541 tcagtgccca acagccgagc ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc
     3601 gccgtggcaa gaactaacca ggatttatac aaggaggaga aaatgaaagc catacgggaa
     3661 gcaatagcat gatacaaagg cattaaagca gcgtatccac atagcgtaaa aggagcaaca
     3721 tagttaagaa taccagtcaa tctttcacaa attttgtaat ccagaggttg attatgatat
     3781 cgaattcgcg gccgcggcgc gccgtcgact tacttgtaca gctcgtccat gccgagagtg
     3841 atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
     3901 ttgctcagct tggactggta gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
     3961 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgccgccgtc ctcgatgttg
     4021 tggcggatct tgaagttggc cttgatgccg ttcttctgct tgtcggcggt gatatagacg
     4081 ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
     4141 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
     4201 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg
     4261 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac
     4321 tgcacgccgt agcccagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
     4381 gtgcagatca gcttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
     4441 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
     4501 gtgaacagct cctcgccctt gctcaccatg gtggcgaccg gtagcgctag cttcgaaaga
     4561 tctggatccc agaagcaaat gtaagcaata gatggctctg ccctgacttt tatgcccagc
     4621 ccgagctccg ccggcgagag ccttgcaaca ggtctaatgt gtaaatgtcc tcaaggaact
     4681 cttctactga agtaaaatcc aaaaatgact cttgttaaac ttcagtgatt atgggaaaca
     4741 cacacacaca caaatttagc tctttaaatt caaagcctac aaacatatca atgtgaattg
     4801 cttaaggaaa ggaagacgga tatgaaacct cctgtttccg gtttctcata ggagaaggag
     4861 cccctctctg tgtgctgaga gtctgtgggt ttatgaaaca agaagctgct tacgttttgc
     4921 atgcaccttt gggtacatct agcccatctt tgtgctatgc tagatttctt gagatatgaa
     4981 gaagcctgga tgattacata tgctttaaca cccctgctac cttctcaaga taggtatgta
     5041 ttaaaatatc tcacacactc ccctctgcaa cccctggact ttagccagag gacacacttc
     5101 ctcttcatca
//
