LOCUS       sequence-460595-        5082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0017h_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in specific brain cell types.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5082)
  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 5082)
  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: AiP11518 -
            pAAV-hsA2-AiE0017h_3xC2-minRho-SYFP...
FEATURES             Location/Qualifiers
     source          1..5082
                     /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      1119..1128
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1125..1127,1128..1130,1131..1841)
                     /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(1170..1191)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1431..1450)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1778..1799
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1909..1929)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2116..2227
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2345..2485
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2345..2474
                     /label=AAV2 ITR
     rep_origin      2560..3015
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2647..2666)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2857..2878
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3032..3051)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3151..3173
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3211..3229)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3297..3401
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3402..3470,3471..4262)
                     /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(3620..3639)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4433..5021
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4922..4941
                     /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 gcggccgcaa cgcgtttaga acaatggctg gcccatagta aatgccgtgt
      181 tagtgtgtta gttgctgttc ttccacgtca gaagaggcac agacaaatta ccaccaggtg
      241 gcgctcagag tctgcggagg catcacaaca gccctgaatt tgaatcctgc tctgccactg
      301 cctagttgag accttttact acctgactag ctgtttgtgt attttaggtg tttgtttgag
      361 gctgggctgc tggcatcagc aggcgcccct cctccccacc tcgctaaaca atcattgcac
      421 aaaatatgca aatggtataa ttactgttat ttgttttgct gatataagtg tttgaaatgc
      481 aaatgtcaag tttgggcgcc ttcatttttc caaccctctc acccggacat gagcagagcc
      541 ctcatcacac agactgaggc tgggctgctg gcatcagcag gcgcccctcc tccccacctc
      601 gctaaacaat cattgcacaa aatatgcaaa tggtataatt actgttattt gttttgctga
      661 tataagtgtt tgaaatgcaa atgtcaagtt tgggcgcctt catttttcca accctctcac
      721 ccggacattg ggtgagtaag agtaggtggc aacgaggctg ggctgctggc atcagcaggc
      781 gcccctcctc cccacctcgc taaacaatca ttgcacaaaa tatgcaaatg gtataattac
      841 tgttatttgt tttgctgata taagtgtttg aaatgcaaat gtcaagtttg ggcgccttca
      901 tttttccaac cctctcaccc ggacatgaat tcgatatcat aatcaaccat aggtaccgag
      961 ctcgggattc agccgggagc ttagggaggg gaggtcactt cataagggcc tgggggggga
     1021 gttggagcca cgagtcgtcc agccggagcc ccgtgtggct gagctccggc ctcagaagca
     1081 tccccgggtt ggatccttcg aagctagcgc taccggtcgc caccatggtg agcaagggcg
     1141 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1201 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     1261 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
     1321 gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     1381 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     1441 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1501 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1561 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     1621 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     1681 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     1741 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1801 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagtcgac atcataatca
     1861 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     1921 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     1981 tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc
     2041 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggc
     2101 tcgagagatc ttcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2161 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2221 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2281 gcaaggggga ggattgggaa gacaatagca ggcatgagat ctcacgtgcg gaccgagcgg
     2341 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     2401 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     2461 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     2521 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     2581 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     2641 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2701 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2761 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2821 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2881 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     2941 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgggaatttt aacaaaatat
     3001 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3061 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3121 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3181 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     3241 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     3301 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3361 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3421 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3481 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3541 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3601 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     3661 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3721 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3781 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3841 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3901 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3961 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4021 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4081 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4141 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4201 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4261 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4321 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4381 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4441 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4501 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4561 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4621 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4681 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4741 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4801 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4861 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4921 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4981 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5041 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
