LOCUS       sequence-470495-        4644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0564h 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 4644)
  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 4644)
  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: AiP12680 - pAAV-AiE0564h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(675..1466,1467..1535))
                     /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     1298..1317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1536..1640)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1708..1726
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1764..1786)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1886..1905
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1922..2377)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2059..2080)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2271..2290
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2452..2592
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2463..2592
                     /label=AAV2 ITR
     polyA_signal    2710..2821
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3008..3028
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3121..3831,3832..3834,3835..3837))
                     /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(3163..3184)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3512..3531
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3771..3792
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3834..3843
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4369..4498
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4560..4644,1..504))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(join(4640..4644,1..15))
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
       61 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
      121 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
      181 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
      241 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
      301 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
      361 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
      421 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
      481 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
      541 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
      601 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
      661 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
      721 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
      781 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
      841 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
      901 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
      961 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     1021 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     1081 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     1141 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     1201 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     1261 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     1321 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     1381 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     1441 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     1501 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     1561 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     1621 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     1681 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     1741 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     1801 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     1861 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     1921 aaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     1981 ttttttaacc aatagaccga aatcggcaaa atcccttata aatcaaaaga atagcccgag
     2041 atagagttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2101 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2161 aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2221 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2281 gcgaaaggag cgggcgctaa ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2341 acacccgccg cgcttaatgc gccgctacag ggcgcgtact atggttgctt tgacgtatgc
     2401 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc ccctgcaggc
     2461 agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct
     2521 ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca
     2581 ctaggggttc ctgcggccgc tcggtccgca cgtgagatct catgcctgct attgtcttcc
     2641 caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc
     2701 agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc
     2761 agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca
     2821 gtcgaagatc tctcgagcca cggaattgtc agtgcccaac agccgagccc ctgtccagca
     2881 gcgggcaagg caggcggcga tgagttccgc cgtggcaaga actaaccagg atttatacaa
     2941 ggaggagaaa atgaaagcca tacgggaagc aatagcatga tacaaaggca ttaaagcagc
     3001 gtatccacat agcgtaaaag gagcaacata gttaagaata ccagtcaatc tttcacaaat
     3061 tttgtaatcc agaggttgat tatgatatcg aattcgcggc cgcggcgcgc cgtcgactta
     3121 cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac
     3181 catgtgatcg cgcttctcgt tggggtcttt gctcagcttg gactggtagc tcaggtagtg
     3241 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
     3301 gagctgcacg ccgccgtcct cgatgttgtg gcggatcttg aagttggcct tgatgccgtt
     3361 cttctgcttg tcggcggtga tatagacgtt gtggctgttg tagttgtact ccagcttgtg
     3421 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
     3481 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa
     3541 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
     3601 catgtggtcg gggtagcggg cgaagcactg cacgccgtag cccagggtgg tcacgagggt
     3661 gggccagggc acgggcagct tgccggtggt gcagatcagc ttcagggtca gcttgccgta
     3721 ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc
     3781 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggt
     3841 ggcgaccggt agcgctagct tcgaaagatc tggatcccag aagcaaatgt aagcaataga
     3901 tggctctgcc ctgactttta tgcccagccc gagctcaaaa aggccctcca atccatgcat
     3961 gcctctgcgg gggctgtgca cccagtgagg tgaggggtca tgcctcagct gttccctgtc
     4021 ttcctctggg gtgcggcgac cacaaccagg gcttagatgt tgtctgggcc agtgtggctg
     4081 gagatgtaac tggtgacaag cgtcggggag cccagaggct agggctggtc tggggcaggc
     4141 gccagggcta tttctgggtg gaaggagaag ggggacccag ctgggctaat gactgggtga
     4201 cctggaactt ccgccttctc tctgttatgt tcccatctat ataaacttgt cagcatggac
     4261 cctaggtagc aacgttctcc aggcacagtg ggggaagagc gcaaggcgtc tgactcacct
     4321 cccagaagac ttgggttcaa atcccagcct tcaaacgcgt gcggccgcag gaacccctag
     4381 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4441 aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcaggac
     4501 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4561 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4621 cgaaacccga caggactata aaga
//
