LOCUS       sequence-461874-        5620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0948m 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 5620)
  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 5620)
  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: AiP13300 -
            pAAV-AiE0948m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1086..1095
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1092..1094,1095..1097,1098..1808)
                     /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(1398..1417)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1745..1766
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1818..1874
                     /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             1887..1952
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1983..2360
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2447..2467)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2654..2765
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2883..3023
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2883..3012
                     /label=AAV2 ITR
     rep_origin      3098..3553
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3185..3204)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3395..3416
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3570..3589)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3689..3711
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3749..3767)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3835..3939
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3940..4008,4009..4800)
                     /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(4158..4177)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4971..5559
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5460..5479
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtggctac ttgaaatacc tttctaactc agtaactaac
      181 taactaatat taaaatgaag ttttgataca gaattgaagg actcaaacat gttcacttca
      241 ttttttactt tcagcttaac accatttatg tgaaatttgg ctagataaat acataccagg
      301 ttgagttctg gagaaggtgg gtagcaggcc tgcttattag accaggtgtg tgggctacca
      361 tcaaaggctt ggcaacttcc tccttttcag aggtctgcat gggtagccaa atactctaga
      421 gcccaggctc atggctggcc tatactgttt cagtgcccaa gcagctgaag tgcagctgct
      481 gtgatctgcc tataattgca gaaaagcatt gttcaaattg agccaaaggg aagaaactct
      541 tactgatttt aaccccccaa acccagaaga tataattctg atgtacctgg aaaatatatc
      601 tacgtaggta agacctggca gggaacaatc ctactgagct tatgtgcctt cattctgtgg
      661 cttggcccca tacttctatg cctatataat tatattaatg gatttcctcc ctttgggtat
      721 gtatccatac attgctcaac acttagagtt acaatggaat ccattcaact attctgagtg
      781 ggtcatggcc ctgctgtttg tttttctatc attactggaa ttatttgggt tccaccacat
      841 gggtacttgg acccacagtg tgatagcacg agtgtaaccc aattaagtta cttgtactag
      901 aaacacaaaa gctatgctca gatctctcct cctgataata ttgcaaagaa tgttccctaa
      961 caagctttgg aacctaaatt ccttgagcct gtgagctcgg gctgggcata aaagtcaggg
     1021 cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac
     1081 cggtcgccac catggtctcc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1141 tcgagctgga cggcgatgtc aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1201 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
     1261 cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg
     1321 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1381 gcaccatctt cttcaaagac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1441 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1501 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
     1561 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
     1621 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1681 ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg
     1741 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1801 tgtacaaggg ctctggtgct accaacttct cactgttgaa acaggcaggg gatgtagagg
     1861 agaatccagg gcctggtgct agtggagact acaaagacca tgacggagat tataaagatc
     1921 atgacatcga ttacaaggat gacgatgaca agtccggact cagatctgga ggctccggag
     1981 gcccagagcc agcgaagtct gctcccgccc cgaaaaaggg ctccaagaag gcggtgacta
     2041 aggcgcagaa gaaaggcggc aagaagcgca agcgcagccg caaggagagc tattccatct
     2101 atgtgtacaa ggttctgaag caggtccacc ctgacaccgg catttcgtcc aaggccatgg
     2161 gcatcatgaa ttcgtttgtg aacgacattt tcgagcgcat cgcaggagag gcttcccgcc
     2221 tggcgcatta caacaagcgc tcgaccatca cctcccggga gatccagacg gccgtgcgcc
     2281 tgctgctgcc tggggagttg gccaagcacg ccgtgtccga gggtactaag gccatcacca
     2341 agtacaccag cgctaagtaa tgaggcgcgc cgcggccgcg aattcgatat cataatcaac
     2401 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2461 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2521 tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg
     2581 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc
     2641 gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2701 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2761 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2821 aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc
     2881 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     2941 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     3001 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     3061 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     3121 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     3181 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3241 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3301 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3361 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3421 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     3481 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3541 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3601 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3661 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3721 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     3781 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3841 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3901 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3961 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4021 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4081 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4141 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4201 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4261 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4321 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4381 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4441 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4501 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4561 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4621 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4681 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4741 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4801 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4861 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4921 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4981 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5041 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5101 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5161 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5221 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5281 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5341 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5401 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5461 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5521 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5581 ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
