LOCUS       sequence-325644-        4825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0064h is an enhancer sequence designed to drive AAV-mediated 
            transgene expression in Pvalb+ and Sst+ cortical interneurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4825)
  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 4825)
  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: AiP11408-pAAV-AiE0064h-minBG-SYFP2-WPRE3-BGHpA...
FEATURES             Location/Qualifiers
     source          1..4825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      819..828
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(825..827,828..830,831..1541)
                     /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(870..891)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1131..1150)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1478..1499
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1634..1654)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1841..1952
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2070..2210
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2070..2199
                     /label=AAV2 ITR
     rep_origin      2285..2740
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2372..2391)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2582..2603
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2757..2776)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2876..2898
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2936..2954)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3022..3126
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3127..3195,3196..3987)
                     /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(3345..3364)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4158..4746
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4647..4666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(4808..4825,1..112)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
       61 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc
      121 acgcgtatga actcttaaat ctcaaatgtg tttgccctag ctgctttgac taaccccctc
      181 ttctatttca gttatgcggc aagttgcata ctcaggtgcc ccttctgact acttgaatac
      241 tttccctgtg atgtaagaag tgttttcaat tggtaaagtt gtggtatata attacaattg
      301 aactctcttg tacttgcctc ttttacaaaa attctctcct agcagaacgt agtgtgagtc
      361 atctacacag ctgtttttct gattattgga attttctttt gacatgaagg aagtatctca
      421 ttgacagaac tgcgttgtga aggagtgcta actgtagcat aaaatacaaa attggatttt
      481 tagattgcaa aatacagtaa agctttgaaa agtatttggc atgacattta actcaataca
      541 ttttgcctaa aaaatattag ccaagaaccc ctatcaactt gtttttgaat aaacttctgt
      601 atggacctta aaattcatgc tgagtttgac cgcattttct tgcactggta gcattttccc
      661 tctgagtcat cctcatttcc ttctactttc tcacatgact aggttaagat aactcatgta
      721 tttgcgagct cgggctgggc ataaaagtca gggcagagcc atctattgct tacatttgct
      781 tctgggatcc agatctttcg aagctagcgc taccggtcgc caccatggtg agcaagggcg
      841 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      901 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      961 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
     1021 gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     1081 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     1141 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1201 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1261 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     1321 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     1381 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     1441 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1501 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagtcgac ggcgcgccgc
     1561 ggccgcgaat tcgatatcat aatcaacctc tggattacaa aatttgtgaa agattgactg
     1621 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     1681 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag
     1741 ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     1801 tgttgggcac tgacaattcc gtggctcgag agatcttcga ctgtgccttc tagttgccag
     1861 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     1921 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     1981 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2041 gagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc
     2101 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     2161 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     2221 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     2281 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2341 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     2401 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2461 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     2521 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2581 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     2641 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     2701 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     2761 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     2821 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     2881 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     2941 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     3001 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3061 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3121 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3181 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3241 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3301 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3361 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3421 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3481 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3541 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3601 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3661 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3721 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     3781 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3841 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3901 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3961 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4021 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4081 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4141 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4201 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4261 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4321 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4381 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4441 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4501 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4561 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4621 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4681 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4741 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4801 cacatgtcct gcaggcagct gcgcg
//
