LOCUS       sequence-470624-        5153 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0967m 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 5153)
  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 5153)
  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: AiP13304 -
            pAAV-AiE0967m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      55..510
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(142..161)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     352..373
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(527..546)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     646..668
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(706..724)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        792..896
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(897..965,966..1757)
                     /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(1115..1134)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1928..2516
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2417..2436
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2578..2707
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3196..3205
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3202..3204,3205..3207,3208..3918)
                     /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(3508..3527)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3855..3876
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3928..3984
                     /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             3997..4062
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4093..4470
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(4557..4577)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4764..4875
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4993..5133
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4993..5122
                     /label=AAV2 ITR
ORIGIN
        1 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
       61 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
      121 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
      181 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
      241 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
      301 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
      361 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
      421 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
      481 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
      541 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
      601 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
      661 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
      721 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
      781 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      841 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      901 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
      961 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1021 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1081 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     1141 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1201 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1261 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     1321 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1381 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1441 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1501 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     1561 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     1621 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     1681 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1741 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     1801 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     1861 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     1921 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     1981 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2041 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     2101 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2161 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2221 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2281 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     2341 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     2401 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2461 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     2521 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct
     2581 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt
     2641 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
     2701 ggttcctgcg gccgcacgcg taggcaacct cagaagctgt catcctctct caactccagc
     2761 ccacgggcac tgcgcttgcc ctggaggaac acttgaaatt cagttctcat ggagggaact
     2821 acttaaaatc agaaaggagg taaaaacccc agctcagggg ttccaagaag agacaaaccc
     2881 ttctcctctc gatccagaat caacaaagct gagtgaggag ttgctgggga ggaatccagc
     2941 cccttgcaaa aagctactgg ctgagcagca gcctgtggcc gctgtctcgg gcagccaagc
     3001 tcagaatgag cacagcctgt atccctggct gcagcgggag aagctaaagt gagctcagtg
     3061 cactgagggg tcctgatacc agcaccggct tctgaaggca gcgagctcgg gctgggcata
     3121 aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag
     3181 ctagcgctac cggtcgccac catggtctcc aagggcgagg agctgttcac cggggtggtg
     3241 cccatcctgg tcgagctgga cggcgatgtc aacggccaca agttcagcgt gtccggcgag
     3301 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
     3361 ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc
     3421 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     3481 gtccaggagc gcaccatctt cttcaaagac gacggcaact acaagacccg cgccgaggtg
     3541 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3601 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     3661 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     3721 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     3781 gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac
     3841 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3901 atggacgagc tgtacaaggg ctctggtgct accaacttct cactgttgaa acaggcaggg
     3961 gatgtagagg agaatccagg gcctggtgct agtggagact acaaagacca tgacggagat
     4021 tataaagatc atgacatcga ttacaaggat gacgatgaca agtccggact cagatctgga
     4081 ggctccggag gcccagagcc agcgaagtct gctcccgccc cgaaaaaggg ctccaagaag
     4141 gcggtgacta aggcgcagaa gaaaggcggc aagaagcgca agcgcagccg caaggagagc
     4201 tattccatct atgtgtacaa ggttctgaag caggtccacc ctgacaccgg catttcgtcc
     4261 aaggccatgg gcatcatgaa ttcgtttgtg aacgacattt tcgagcgcat cgcaggagag
     4321 gcttcccgcc tggcgcatta caacaagcgc tcgaccatca cctcccggga gatccagacg
     4381 gccgtgcgcc tgctgctgcc tggggagttg gccaagcacg ccgtgtccga gggtactaag
     4441 gccatcacca agtacaccag cgctaagtaa tgaggcgcgc cgcggccgcg aattcgatat
     4501 cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     4561 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     4621 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     4681 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     4741 tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     4801 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     4861 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4921 gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga
     4981 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     5041 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     5101 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat ttt
//
