LOCUS       sequence-470750-        5429 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0923m 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 5429)
  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 5429)
  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: AiP14095 -
            pAAV-AiE0923m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(116..907,908..976))
                     /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     739..758
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(977..1081)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1149..1167
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1205..1227)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1327..1346
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1363..1818)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1500..1521)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1712..1731
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     misc_feature    1887..1914
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     misc_feature    1915..1942
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     repeat_region   1949..2089
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1960..2089
                     /label=AAV2 ITR
     polyA_signal    2207..2318
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2505..2525
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2612..2989)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(3020..3085)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(3098..3154)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(join(3164..3874,3875..3877,3878..3880))
                     /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(3206..3227)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3555..3574
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      3877..3886
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4538..4667
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     misc_feature    4674..4701
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     misc_feature    4702..4729
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     rep_origin      complement(4786..5374)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4866..4885)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
       61 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
      121 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
      181 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
      241 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
      301 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
      361 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
      421 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
      481 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
      541 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
      601 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
      661 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
      721 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
      781 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
      841 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
      901 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
      961 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     1021 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     1081 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     1141 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     1201 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     1261 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     1321 actatgcggc atcagagcag attgtactga gagtgcacca taaaattgta aacgttaata
     1381 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caatagaccg
     1441 aaatcggcaa aatcccttat aaatcaaaag aatagcccga gatagagttg agtgttgttc
     1501 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1561 ccgtctatca gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt
     1621 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1681 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     1741 aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     1801 cgccgctaca gggcgcgtac tatggttgct ttgacgtatg cggtgtgaaa taccgcacag
     1861 atgcgtaagg agaaaatacc gcatcatatc agcacacaat agtccattat acgcgcgtat
     1921 aatgggcaat tgtgtgctga taggcgcccc tgcaggcagc tgcgcgctcg ctcgctcact
     1981 gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc
     2041 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgctcg
     2101 gtccgcacgt gagatctcat gcctgctatt gtcttcccaa tcctccccct tgctgtcctg
     2161 ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg caatttcctc
     2221 attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag gcacggggga
     2281 ggggcaaaca acagatggct ggcaactaga aggcacagtc gaagatctct cgagccacgg
     2341 aattgtcagt gcccaacagc cgagcccctg tccagcagcg ggcaaggcag gcggcgatga
     2401 gttccgccgt ggcaagaact aaccaggatt tatacaagga ggagaaaatg aaagccatac
     2461 gggaagcaat agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag
     2521 caacatagtt aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattat
     2581 gatatcgaat tcgcggccgc ggcgcgcctc attacttagc gctggtgtac ttggtgatgg
     2641 ccttagtacc ctcggacacg gcgtgcttgg ccaactcccc aggcagcagc aggcgcacgg
     2701 ccgtctggat ctccctggag gtgatggtcg agcgcttgtt gtaatgcgcc aggcgggaag
     2761 cctcacctgc gatgcgctcg aaaatgtcgt tcacaaacga gttcatgatg cccatggcct
     2821 tggacgaaat gccggtgtca gggtggacct gcttcagaac cttatacaca tagatggaat
     2881 agctctcctt gcggctgcgc ttgcgcttct tgccgccttt cttctgcgcc ttagtcaccg
     2941 ccttcttgga gccctttttc ggggcgggag cagacttcgc tggctctggg cctccggagc
     3001 ctccagatct gagtccggac ttgtcatcgt catccttgta atcgatgtca tgatctttat
     3061 aatctccgtc atggtctttg tagtctccac tagcaccagg ccctggattc tcctctacat
     3121 cccctgcctg tttcaacagt gagaagttgg tagcaccaga gcccttgtac agctcgtcca
     3181 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     3241 cgttggggtc tttgctcagc ttggactggt agctcaggta gtggttgtcg ggcagcagca
     3301 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt
     3361 cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg
     3421 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     3481 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     3541 tcacctcggc gcgggtcttg tagttgccgt cgtctttgaa gaagatggtg cgctcctgga
     3601 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     3661 gggcgaagca ctgcacgccg tagcccaggg tggtcacgag ggtgggccag ggcacgggca
     3721 gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     3781 cctcgccgga cacgctgaac ttgtggccgt tgacatcgcc gtccagctcg accaggatgg
     3841 gcaccacccc ggtgaacagc tcctcgccct tggagaccat ggtggcgacc ggtagcgcta
     3901 gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat agatggctct gccctgactt
     3961 ttatgcccag cccgagctcc attctcaaat ctcttttagt gggaggaggg ggatctatta
     4021 tatgaaagta tacattctaa cttcttgtaa gctgtggaga aagaatgaga gaaggaaagg
     4081 aaaaaggaaa caaaaatttg agtttctatg aaatgggaga attctgttac atgtgactaa
     4141 cttaattctt aacttcttta actactatgt gaagtaaatg ctgccttagc tgtgaatata
     4201 tatatataaa agtgttgccc tttaacacag tctgtattta gaagagctga aattcaattc
     4261 ctgagtcata tggatttcaa actttcaacc ttccacttca taagaatgaa acatcaaaag
     4321 agaaaaattg aaacttgtct tccagtgcca tgcctttggg cataaaataa taaaatatga
     4381 ctaagaaata gagctacatg caagtggggt gacaacagat tcaacccata attgatcttt
     4441 ttctaaaaca cacacacaca cacacacaca cacacacaca cacacacaca cacacacacc
     4501 tattttattt gaaatgttaa ctcacgcgtg cggccgcagg aacccctagt gatggagttg
     4561 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     4621 cgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggaca tgttatcagc
     4681 acacaatagt ccattatacg cgcgtataat gggcaattgt gtgctgatat gagcaaaagg
     4741 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4801 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4861 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4921 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4981 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5041 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5101 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5161 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5221 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5281 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5341 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5401 gtctgacgct cagtggaacg aaaactcac
//
