LOCUS       sequence-466350-        4863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3061m 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 4863)
  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 4863)
  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: AiP20020 - pAAV-AiE3061m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      454..463
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(460..462,463..465,466..1176)
                     /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(505..526)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(766..785)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1113..1134
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1269..1289)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1476..1587
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1705..1845
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1705..1834
                     /label=AAV2 ITR
     rep_origin      1920..2375
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2007..2026)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2217..2238
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2392..2411)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2511..2533
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2571..2589)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2657..2761
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2762..2830,2831..3622)
                     /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(2980..2999)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3793..4381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4282..4301
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4443..4572
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctcttattca tttgttaata atgtgttgtt tttttgggaa atagccctca ctctgttggc
       61 tcagaagagg aatccaaggg aaatgtcatg ctgaatcctg tgctagaatt caaagcaaca
      121 cgtgtgcagg agtgatctgg tcaaacaagg atatggtttc ctaaatgggt ggatccagac
      181 agaaggtgga ccatggatgg tagatgctgc ttcaaatatg gcttctgatg agcacattga
      241 atggagcaag ggatcacagg acccacggac tctggaggac acatagctgt gacctctgaa
      301 agggaatcaa gggaggggat ggtcctacaa atcctgaccc tgtggcatac aacgccggcg
      361 gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg
      421 gatccagatc tttcgaagct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag
      481 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      541 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg
      601 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac
      661 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
      721 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
      781 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
      841 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
      901 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
      961 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     1021 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag
     1081 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1141 gccgggatca ctctcggcat ggacgagctg tacaagtaag tcgacggcgc gccgcggccg
     1201 cgaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     1261 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     1321 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt
     1381 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     1441 ggcactgaca attccgtggc tcgagagatc ttcgactgtg ccttctagtt gccagccatc
     1501 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     1561 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     1621 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat
     1681 ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     1741 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     1801 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     1861 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     1921 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1981 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2041 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2101 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     2161 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2221 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     2281 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2341 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     2401 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     2461 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     2521 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     2581 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     2641 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2701 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2761 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2821 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2881 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     2941 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3001 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3061 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3121 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3181 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3241 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3301 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3361 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3421 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3481 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3541 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3601 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3661 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3721 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3781 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3841 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3901 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3961 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4021 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4081 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4141 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4201 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4261 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4321 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4381 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4441 gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct
     4501 ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca
     4561 ctaggggttc ctgcggccgc acgcgtagac catgggatga aggagtgtgg ctggccagtg
     4621 ggtggggtga ggggccttac cagggttctg agaatgagtc ctctatctct gaggacctgg
     4681 cactgacctg tgtggggcag cccttggcca gtgaatagtc ctcactttgg tgactttctt
     4741 ggctctgttg gatataagag gtctgaaagt ttccccaaga gccttgtgaa ttaaatacat
     4801 ttgcgatttg tattctaaac aagcttgcgt ggactaaata tgctggctgt attgttttct
     4861 tgg
//
