LOCUS       sequence-470650-        5433 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0892m 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 5433)
  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 5433)
  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: AiP13409 -
            pAAV-AiE0892m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5433
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     239..259
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(346..723)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(754..819)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(832..888)
                     /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(898..1608,1609..1611,1612..1614))
                     /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(940..961)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1289..1308
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      1611..1620
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2389..2518
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2580..3168)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2660..2679)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3339..4130,4131..4199))
                     /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     3962..3981
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4200..4304)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4372..4390
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4428..4450)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4550..4569
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4586..5041)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4723..4744)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4935..4954
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5116..5256
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5127..5256
                     /label=AAV2 ITR
     polyA_signal    join(5374..5433,1..52)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaagat
       61 ctctcgagcc acggaattgt cagtgcccaa cagccgagcc cctgtccagc agcgggcaag
      121 gcaggcggcg atgagttccg ccgtggcaag aactaaccag gatttataca aggaggagaa
      181 aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag cgtatccaca
      241 tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa ttttgtaatc
      301 cagaggttga ttatgatatc gaattcgcgg ccgcggcgcg cctcattact tagcgctggt
      361 gtacttggtg atggccttag taccctcgga cacggcgtgc ttggccaact ccccaggcag
      421 cagcaggcgc acggccgtct ggatctcccg ggaggtgatg gtcgagcgct tgttgtaatg
      481 cgccaggcgg gaagcctctc ctgcgatgcg ctcgaaaatg tcgttcacaa acgaattcat
      541 gatgcccatg gccttggacg aaatgccggt gtcagggtgg acctgcttca gaaccttgta
      601 cacatagatg gaatagctct ccttgcggct gcgcttgcgc ttcttgccgc ctttcttctg
      661 cgccttagtc accgccttct tggagccctt tttcggggcg ggagcagact tcgctggctc
      721 tgggcctccg gagcctccag atctgagtcc ggacttgtca tcgtcatcct tgtaatcgat
      781 gtcatgatct ttataatctc cgtcatggtc tttgtagtct ccactagcac caggccctgg
      841 attctcctct acatcccctg cctgtttcaa cagtgagaag ttggtagcac cagagccctt
      901 gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat
      961 gtgatcgcgc ttctcgttgg ggtctttgct cagcttggac tggtagctca ggtagtggtt
     1021 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag
     1081 ctgcacgccg ccgtcctcga tgttgtggcg gatcttgaag ttggccttga tgccgttctt
     1141 ctgcttgtcg gcggtgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc
     1201 caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt
     1261 gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtctt tgaagaagat
     1321 ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat
     1381 gtggtcgggg tagcgggcga agcactgcac gccgtagccc agggtggtca cgagggtggg
     1441 ccagggcacg ggcagcttgc cggtggtgca gatcagcttc agggtcagct tgccgtaggt
     1501 ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgttgacat cgccgtccag
     1561 ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttggaga ccatggtggc
     1621 gaccggtagc gctagcttcg aaagatctgg atcccagaag caaatgtaag caatagatgg
     1681 ctctgccctg acttttatgc ccagcccgag ctcagcaaag tcctggaggg gtaaattaaa
     1741 aagctggctt ggttttctag tcccatcctg gggtctccta tttataatta gtgtctacga
     1801 taaaattagc catcaaatct tttagggctc caaaatattg gcctgaggtt tcatttacat
     1861 tgtatttctg gcagtttgaa taatgttcca aactagtcgc attgattaaa tagacccatt
     1921 ttgtgactta aatgttttaa catctgtatt gctcaaagat ctttctgaat cagtacattt
     1981 ctacagatac aaaaataaca tttgtcccaa attttgagaa tctattagaa agccttgact
     2041 gaaagcacat tcttgtataa gcagtgcccg caagaatgcc caaataactg tgaccttgag
     2101 gactttggaa gcaggactgc cattttgctt tttttgtcaa gttccccctc ccccccacag
     2161 tcactcctgc agctggcttc ttcttctctc atgcagtcct gttaaatatg tatggcctgg
     2221 ttagttttat tttcaaagaa ataacatttt ttctcgtcct tgttacttgt aacatataat
     2281 cagctgaatt gcaccgttat agcccaccgg gtgcttcttc ctctcctgtc agatttgttt
     2341 ttttctctgt atcccaagaa atctttattt tgcaacgcgt gcggccgcag gaacccctag
     2401 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     2461 aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcaggac
     2521 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2581 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2641 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2701 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2761 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2821 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2881 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2941 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3001 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3061 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3121 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3181 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3241 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3301 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3361 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3421 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3481 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3541 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3601 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3661 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3721 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3781 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3841 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3901 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3961 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4021 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4081 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4141 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4201 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4261 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4321 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     4381 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     4441 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     4501 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     4561 cagattgtac tgagagtgca ccataaaatt gtaaacgtta atattttgtt aaaattcgcg
     4621 ttaaattttt gttaaatcag ctcatttttt aaccaataga ccgaaatcgg caaaatccct
     4681 tataaatcaa aagaatagcc cgagatagag ttgagtgttg ttccagtttg gaacaagagt
     4741 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4801 ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca
     4861 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4921 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctaaggcgct ggcaagtgta
     4981 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     5041 tactatggtt gctttgacgt atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     5101 accgcatcag gcgcccctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
     5161 caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca
     5221 gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgctcggtc cgcacgtgag
     5281 atctcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc
     5341 cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa
     5401 aggacagtgg gagtggcacc ttccagggtc aag
//
