LOCUS       sequence-325645-        4940 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0359h is an enhancer sequence designed to drive AAV-mediated 
            transgene expression in Pvalb+ cortical interneurons and cerebellar 
            Purkinje cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4940)
  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 4940)
  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: 
            AiP12045-pAAV-3xSP10ins-AiE0359h-minRho-SYFP2-WPRE3-BGHpA (Alias: 
            CN2045)
FEATURES             Location/Qualifiers
     source          1..4940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     164..185
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(320..340)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    527..638
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   756..896
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   756..885
                     /label=AAV2 ITR
     rep_origin      971..1426
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1058..1077)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1268..1289
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1443..1462)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1562..1584
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1622..1640)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1708..1812
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1813..1881,1882..2673)
                     /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(2031..2050)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2844..3432
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3333..3352
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3494..3623
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      4445..4454
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4451..4453,4454..4456,4457..4940,1..227)
                     /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(4496..4517)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4757..4776)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
       61 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
      121 accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
      181 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa gtcgacggcg
      241 cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat
      301 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
      361 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
      421 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
      481 ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt
      541 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
      601 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
      661 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
      721 aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
      781 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
      841 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
      901 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
      961 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1021 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     1081 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     1141 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     1201 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     1261 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     1321 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     1381 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     1441 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     1501 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     1561 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     1621 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     1681 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1741 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1801 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     1861 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     1921 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     1981 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2041 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2101 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2161 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2221 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2281 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     2341 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     2401 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     2461 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     2521 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     2581 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     2641 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     2701 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     2761 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2821 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2881 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2941 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3001 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3061 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3121 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3181 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3241 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3301 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3361 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3421 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3481 tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg
     3541 tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc
     3601 caactccatc actaggggtt cctgcggccg cacgcgccgg taccgaagct acccctaaca
     3661 cactattcta cacacagaaa atgctcttca ctaggaagct acccctaaca cactattcta
     3721 cacacagaaa atgctcttca ctaggaagct acccctaaca cactattcta cacacagaaa
     3781 atgctcttca ctagacgcgt tcacagctcc ttggtctgaa ctggtaccca ggatcaagat
     3841 gtccctccca tgattctaat ctccagcccc cttacaccat agtcacttcc aaactctgga
     3901 tttcttaaca ttgcagtttg cttccaaaaa aaaattaaaa aaaaacctca tctgggcatt
     3961 agaagtcatt ctgaagaggc tgtcctatat ctcagtgcaa tttcctactt atccactccc
     4021 gactataccc tgctttcatc agattgcaac tgacctaatt tgtaaagact tcagtgaaaa
     4081 gagcctcatt aaaagccttc atctcttttg cagcctctct ttcctccctc aaatgatcca
     4141 ggtcagaatg gcccaataaa aatagctggg aaaattacaa agtcactcaa gactctgcca
     4201 ggagtagaga ttttaagaat tagtgacaga catatactgt ctaggagtag ggtgggagtg
     4261 ggttgcatgg cttagggaca tagagagctc gattcagccg ggagcttagg gaggggaggt
     4321 cacttcataa gggcttgggg ggggagttgg agccacgagt cgtccagccg gagccccgtg
     4381 tggctgtgct ccggcctcag aagcatcccc ggatccagat ctttcgaagc tagcgctacc
     4441 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     4501 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     4561 tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc
     4621 ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc gctaccccga
     4681 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     4741 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     4801 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     4861 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa
     4921 gcagaagaac ggcatcaagg
//
