LOCUS       sequence-470574-        4962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0496m 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 4962)
  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 4962)
  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: AiP12939 - pAAV-AiE0496m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      375..384
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(381..383,384..386,387..1097)
                     /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(426..447)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(687..706)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1034..1055
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1190..1210)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1397..1508
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1626..1766
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1626..1755
                     /label=AAV2 ITR
     rep_origin      1841..2296
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1928..1947)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2138..2159
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2313..2332)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2432..2454
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2492..2510)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2578..2682
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2683..2751,2752..3543)
                     /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(2901..2920)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3714..4302
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4203..4222
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4364..4493
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcccgcctgt gggcgctgtc ccgggctgcg agggccgcga gcgcaccgac agacgaaccg
       61 accgacggag gactggctgc gggccggaca gacggggtag cgcagggaac tgggatgagg
      121 cgggacagcg acatggcaag ccacatacaa cagccaggcg ggcacgggaa ccccggccct
      181 gcgccctcgc cttccccggg ccctggtccc ggcccgggcg cctcggagcg ggtggcactc
      241 aagaaagaga tcgggctggt gagcgcttgc accatcatca tgagctcggg ctgggcataa
      301 aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat ctttcgaagc
      361 tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
      421 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
      481 gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc
      541 tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc
      601 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
      661 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
      721 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
      781 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
      841 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg
      901 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
      961 tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa gaccccaacg
     1021 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1081 tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc gcgaattcga tatcataatc
     1141 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     1201 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     1261 ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg
     1321 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     1381 ctcgagagat cttcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1441 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1501 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     1561 agcaaggggg aggattggga agacaatagc aggcatgaga tctcacgtgc ggaccgagcg
     1621 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     1681 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     1741 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     1801 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     1861 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     1921 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     1981 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2041 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2101 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2161 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     2221 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     2281 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     2341 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     2401 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     2461 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     2521 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     2581 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2641 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2701 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2761 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2821 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2881 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     2941 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3001 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3061 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3121 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3181 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     3241 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3301 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3361 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3421 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3481 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3541 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3601 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3661 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3721 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3781 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3841 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3901 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3961 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4021 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4081 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4141 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4201 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4261 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4321 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc
     4381 gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
     4441 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg
     4501 cacgcgtaga ggccagggag ctatcaaaaa gtcttcatgg agaccgtgcc actgtcaggc
     4561 taattgccgc taggcggtgg ttttcaacct gtgcaatgcc cacccttcac aaatggggct
     4621 tgagggattt agaaagtaca ggctggggca tgcagcaccg cctaccccag cacgtcttgc
     4681 tcacactcct tgagcgcagg gacagcctgg tctttgagct cttctccccc acctcaccca
     4741 agtctgggtg tagccccttc gttcactcaa ctaggaagtc ctttctaagt ctgatcccag
     4801 tcccctcctg tctcccctga ctagggttgg gggtgggggg gaagggggag ttccctttaa
     4861 gggcggtggg ggagggggct gccggccgtc cctccttccg tccctccctg gatccagccg
     4921 cccgggggcc cccgggggcc cgggccccgt gccagcgcat gc
//
