LOCUS       sequence-470440-        4606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0402m 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 4606)
  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 4606)
  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: AiP12092 - pAAV-AiE0402m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   339..468
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(530..1118)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(610..629)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1289..2080,2081..2149))
                     /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     1912..1931
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2150..2254)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2322..2340
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2378..2400)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2500..2519
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2536..2991)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2673..2694)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2885..2904
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3066..3206
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3077..3206
                     /label=AAV2 ITR
     polyA_signal    3324..3435
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3622..3642
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3735..4445,4446..4448,4449..4451))
                     /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(3777..3798)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4126..4145
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4385..4406
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4448..4457
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 aagaaccaca actcccgagg gcaatcaatc ccctcagaac actgcccacc caagcctcgc
       61 ctcctctctc cccattccct tccctgacag cctccggcca ccccaggacc tcagagtcgg
      121 ctctggagca cacagacaat gggctgattg ttgtaagatt ctgtcatgct ctttgaaaca
      181 ttcctcatga aaagaaccat acaatgtgca aacagtggca gacagcacag ggtggggcag
      241 gaggaaggca agcaccaaaa agccaccacc aatagcatct agcccagcag ggggacctac
      301 tgtgtgccag gcccttctct aagcacgcgt gcggccgcag gaacccctag tgatggagtt
      361 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
      421 acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcaggac atgtgagcaa
      481 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      541 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
      601 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
      661 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
      721 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
      781 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
      841 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
      901 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
      961 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1021 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1081 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1141 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1201 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     1261 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     1321 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1381 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1441 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1501 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1561 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1621 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1681 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1741 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     1801 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     1861 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     1921 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     1981 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2041 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2101 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2161 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2221 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     2281 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     2341 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     2401 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     2461 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     2521 tgagagtgca ccataaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt
     2581 gttaaatcag ctcatttttt aaccaataga ccgaaatcgg caaaatccct tataaatcaa
     2641 aagaatagcc cgagatagag ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2701 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2761 gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2821 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2881 aggaagggaa gaaagcgaaa ggagcgggcg ctaaggcgct ggcaagtgta gcggtcacgc
     2941 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tactatggtt
     3001 gctttgacgt atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     3061 gcgcccctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg
     3121 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
     3181 ggccaactcc atcactaggg gttcctgcgg ccgctcggtc cgcacgtgag atctcatgcc
     3241 tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc cccagaatag
     3301 aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg
     3361 gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc
     3421 aactagaagg cacagtcgaa gatctctcga gccacggaat tgtcagtgcc caacagccga
     3481 gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc aagaactaac
     3541 caggatttat acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa
     3601 ggcattaaag cagcgtatcc acatagcgta aaaggagcaa catagttaag aataccagtc
     3661 aatctttcac aaattttgta atccagaggt tgattatgat atcgaattcg cggccgcggc
     3721 gcgccgtcga cttacttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg
     3781 aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag cttggactgg
     3841 tagctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc
     3901 tggtagtggt cggcgagctg cacgccgccg tcctcgatgt tgtggcggat cttgaagttg
     3961 gccttgatgc cgttcttctg cttgtcggcg gtgatataga cgttgtggct gttgtagttg
     4021 tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg
     4081 atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg
     4141 tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag
     4201 aagtcgtgct gcttcatgtg gtcggggtag cgggcgaagc actgcacgcc gtagcccagg
     4261 gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat cagcttcagg
     4321 gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg
     4381 tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc
     4441 ttgctcacca tggtggcgac cggtagcgct agcttcgaaa gatctggatc ccagaagcaa
     4501 atgtaagcaa tagatggctc tgccctgact tttatgccca gcccgagctc gggtcccaaa
     4561 tagcttgtgc ataaaaagac ctgaacaaaa ggcctgattc tgacac
//
