LOCUS       sequence-470273-        4872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0064h 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 4872)
  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 4872)
  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: AiP14002 -
            pAAV-AiE0064h-spacer-AiE0022h-minBG...
FEATURES             Location/Qualifiers
     source          1..4872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      906..915
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(912..914,915..917,918..1628)
                     /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(957..978)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1218..1237)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1565..1586
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1721..1741)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1928..2039
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2350..2805
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2437..2456)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2647..2668
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2822..2841)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2941..2963
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3001..3019)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3087..3191
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3192..3260,3261..4052)
                     /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(3410..3429)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4223..4811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4712..4731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttcaat tggtaaagtt gtggtatata attacaattg
      181 aactctcttg tacttgcctc ttttacaaaa attctctcct agcagaacgt agtgtgagtc
      241 atctacacag ctgtttttct gattattgga attttctttt gacatgaagg aagtatctca
      301 ttgacagaac tgcgttgtga aggagtgcta actgtagcat aaaatacaaa attggatttt
      361 tagattgcaa aatacagtaa agctttgggg gcccccagtc ctgtggatac catctgagcc
      421 tgtgcccttt tctctctact cttgagcaag tgcccatcag gagccacatt tgagtcagga
      481 cctgccaaga gcattttcat ctcatcctca gttccccttt tgtctggaac cactcgctat
      541 gagtttctat gagatcgtag ccaggtttga ccaaaccact cctgcacaca tgttgttttg
      601 aaagattagc acagaatcca acacaacatt tcccttggat tcatctttca agccaacaga
      661 gaaagagcta tttccaaaaa aataacacat tattaaccaa tgaataagag ccaagaaaac
      721 aatgtagcca gcatatttat tccaaacagc ttgtttagaa tgcaaattgc aaatacatct
      781 atttccaccc ggctctttgg ggaaccatgc tggagcttgg gctgggcata aaagtcaggg
      841 cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac
      901 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      961 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1021 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
     1081 cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg
     1141 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1201 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1261 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1321 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
     1381 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
     1441 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1501 ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg
     1561 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1621 tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg atatcataat caacctctgg
     1681 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     1741 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     1801 tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc gccgcctgcc
     1861 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gctcgagaga
     1921 tcttcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     1981 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2041 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2101 gaggattggg aagacaatag caggcatgag atctcacgtg cggaccgagc ggccgcagga
     2161 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     2221 gcgaccaaag gtcgcccggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     2281 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     2341 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2401 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     2461 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2521 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     2581 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2641 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     2701 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     2761 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     2821 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     2881 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     2941 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     3001 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     3061 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3121 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3181 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3241 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3301 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3361 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3421 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3481 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3541 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3601 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3661 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3721 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3781 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3841 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3901 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3961 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4021 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4081 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4141 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4201 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4261 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4321 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4381 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4441 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4501 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4561 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4621 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4681 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4741 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4801 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4861 ttgctcacat gt
//
