LOCUS       sequence-466359-        4357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3091m 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 4357)
  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 4357)
  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: AiP20048 - pAAV-AiE3091m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      73..82
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(79..81,82..84,85..795)
                     /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(124..145)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(385..404)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     732..753
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(888..908)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1095..1206
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1324..1464
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1324..1453
                     /label=AAV2 ITR
     rep_origin      1539..1994
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1626..1645)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1836..1857
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2011..2030)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2130..2152
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2190..2208)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2276..2380
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2381..2449,2450..3241)
                     /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     complement(2599..2618)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3412..4000
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3901..3920
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4062..4191
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct ttcgaagcta
       61 gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      121 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      181 gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg
      241 cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc
      301 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      361 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      421 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      481 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc
      541 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac
      601 ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
      661 ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag
      721 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
      781 gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc gaattcgata tcataatcaa
      841 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
      901 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
      961 ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc
     1021 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct
     1081 cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1141 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1201 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1261 caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc
     1321 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     1381 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     1441 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     1501 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     1561 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     1621 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     1681 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     1741 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     1801 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     1861 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     1921 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     1981 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2041 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     2101 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     2161 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     2221 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     2281 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2341 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2401 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2461 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2521 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     2581 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     2641 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     2701 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     2761 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     2821 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     2881 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     2941 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3001 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3061 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3121 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3181 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3241 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3301 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3361 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3421 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3481 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3541 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3601 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3661 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3721 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3781 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3841 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3901 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3961 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4021 tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc
     4081 tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     4141 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca
     4201 cgcgtgccgg gatgtcaaca acccacggaa cctcaggtgg tatcaacccg gaaccctgaa
     4261 gttcactcct ccatgtttta atgtttcctg ggacatagga cctgctgctt ctcccagggc
     4321 gcacaaaccg ccggcggagc tcgggctggg cataaaa
//
