LOCUS       sequence-470631-        5296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0942m 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 5296)
  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 5296)
  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: AiP13313 -
            pAAV-AiE0942m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      515..1103
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1004..1023
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1165..1294
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1926..1935
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1932..1934,1935..1937,1938..2648)
                     /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(2238..2257)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2585..2606
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2658..2714
                     /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             2727..2792
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2823..3200
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(3287..3307)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3494..3605
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3723..3863
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3723..3852
                     /label=AAV2 ITR
     rep_origin      3938..4393
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4025..4044)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4235..4256
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4410..4429)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4529..4551
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4589..4607)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4675..4779
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4780..4848,4849..5296,1..344)
                     /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(4998..5017)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
       61 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
      121 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
      181 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
      241 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
      301 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
      361 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
      421 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
      481 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
      541 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
      601 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
      661 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
      721 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
      781 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
      841 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
      901 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
      961 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     1021 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     1081 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     1141 tttgctggcc ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg
     1201 ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
     1261 gagggagtgg ccaactccat cactaggggt tcctgcggcc gcacgcgtac agtggcccgc
     1321 aagtgcagga agtcaacatc aagcgtgaaa gacagagcaa ttcaagagac tctacagaga
     1381 aatcagaacc cttgaggtca gccaaggttt cccgggacct cgtgctgagt gctaggcgac
     1441 ttaatgtgac tcccagtggg gaagtcaaat gagtgaatga gcagaacagc cctctgattc
     1501 ggtattgctg cagcccatga cttaatcttg ccagaaagaa ctcattcaca gtatccttag
     1561 ctcctagagc acaaagcaat gtaaagagga ttggactggg agtggaagta tttcaggaac
     1621 attttgaata tggccaccca catatatcca tctcctcaca tctcagcaaa ttgtgaaaat
     1681 aaggagtaaa aactcaccat ttgcttcctg acagaaaacc agaccatttc ttaaaaaaaa
     1741 aaaaaaatga agcaagagag gcaggtaatc attcccgttg aagtggtaac acataaaaga
     1801 ggagtgacaa ggagcagaaa ggagattgcc ttgagctcgg gctgggcata aaagtcaggg
     1861 cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac
     1921 cggtcgccac catggtctcc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1981 tcgagctgga cggcgatgtc aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     2041 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
     2101 cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg
     2161 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2221 gcaccatctt cttcaaagac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2281 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2341 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
     2401 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
     2461 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2521 ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg
     2581 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2641 tgtacaaggg ctctggtgct accaacttct cactgttgaa acaggcaggg gatgtagagg
     2701 agaatccagg gcctggtgct agtggagact acaaagacca tgacggagat tataaagatc
     2761 atgacatcga ttacaaggat gacgatgaca agtccggact cagatctgga ggctccggag
     2821 gcccagagcc agcgaagtct gctcccgccc cgaaaaaggg ctccaagaag gcggtgacta
     2881 aggcgcagaa gaaaggcggc aagaagcgca agcgcagccg caaggagagc tattccatct
     2941 atgtgtacaa ggttctgaag caggtccacc ctgacaccgg catttcgtcc aaggccatgg
     3001 gcatcatgaa ttcgtttgtg aacgacattt tcgagcgcat cgcaggagag gcttcccgcc
     3061 tggcgcatta caacaagcgc tcgaccatca cctcccggga gatccagacg gccgtgcgcc
     3121 tgctgctgcc tggggagttg gccaagcacg ccgtgtccga gggtactaag gccatcacca
     3181 agtacaccag cgctaagtaa tgaggcgcgc cgcggccgcg aattcgatat cataatcaac
     3241 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     3301 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     3361 tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg
     3421 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc
     3481 gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3541 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3601 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3661 aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc
     3721 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     3781 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     3841 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     3901 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     3961 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     4021 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4081 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4141 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4201 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4261 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     4321 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4381 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4441 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4501 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4561 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4621 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4681 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4741 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4801 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4861 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4921 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4981 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5041 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5101 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5161 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5221 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5281 aatgaagcca taccaa
//
