LOCUS       sequence-466271-        4739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2040m 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 4739)
  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 4739)
  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: AiP1533 - pAAV-AiE2040m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     166..187
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(341..360)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     460..482
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(520..538)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        606..710
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(711..779,780..1571)
                     /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(929..948)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1742..2330
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2231..2250
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2392..2521
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3142..3151
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3148..3150,3151..3153,3154..3864)
                     /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(3193..3214)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3454..3473)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3801..3822
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3957..3977)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4164..4275
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4393..4533
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4393..4522
                     /label=AAV2 ITR
     rep_origin      join(4608..4739,1..324)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4695..4714)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
       61 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
      121 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
      181 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
      241 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
      301 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
      361 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
      421 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
      481 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
      541 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
      601 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      661 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
      721 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
      781 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
      841 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
      901 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
      961 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1021 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     1081 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     1141 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     1201 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     1261 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     1321 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     1381 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     1441 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     1501 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     1561 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     1621 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     1681 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     1741 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     1801 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     1861 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     1921 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     1981 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2041 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2101 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2161 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2221 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2281 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2341 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc
     2401 agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg
     2461 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
     2521 tgcggccgca cgcgttagaa tgcaggctct ttccccttac ctggcccatt aggagcgcca
     2581 attagtgaaa actggtgcca ttgctgcctt tacaaattaa tgttgggata gccctccacc
     2641 caggaaggca gatatgtctc cttcgtgcct agccccctcc cccctccaca gctctcctct
     2701 taagtgccca cttcctccgg aattatcatc agctgtcttg ttgggagtct gaatagagag
     2761 attaatgagc taataggaaa ttaagactca gaaagcaatt tccagttatc taaatcgagg
     2821 cgtttcatct aatcggttca ggagtgtgga agggagctgc gtgcagagag gccggatgtc
     2881 agcactattc tgcattgtct catcagtacg gactcagctg ccagggtctg cccttgcagt
     2941 gggtttcctg gcttttggtt gtctgtgtcc caccccactt agagtttctt ccatgggggt
     3001 ctaaagcccc tagatctcat ttattggaaa cctttctatc tctgtctcga gctcgggctg
     3061 ggcataaaag tcagggcaga gccatctatt gcttacattt gcttctggga tccagatctt
     3121 tcgaagctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     3181 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     3241 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
     3301 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc
     3361 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     3421 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     3481 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     3541 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     3601 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     3661 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     3721 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac
     3781 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     3841 ctcggcatgg acgagctgta caagtaagtc gacggcgcgc cgcggccgcg aattcgatat
     3901 cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3961 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     4021 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     4081 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     4141 tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     4201 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     4261 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4321 gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga
     4381 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     4441 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     4501 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     4561 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     4621 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4681 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggct
//
