LOCUS       sequence-452978-        5029 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0082h_3xC3 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 5029)
  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 5029)
  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: AiP14484 -
            pAAV-AiE0082h_3xC3-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..5029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     regulatory      1047..1056
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1053..1055,1056..1058,1059..1769)
                     /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(1098..1119)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1359..1378)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1706..1727
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1862..1882)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2069..2180
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2292..2432
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2292..2421
                     /label=AAV2 ITR
     rep_origin      2507..2962
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2594..2613)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2804..2825
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2979..2998)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3098..3120
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3158..3176)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3244..3348
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3349..3417,3418..4209)
                     /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(3567..3586)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4380..4968
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4869..4888
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtccatc tccctctctg cctcccgttt
      181 cccctttact ctttactgtt agttggcctg gggttcctgg gctgaagtgg agtttcctcc
      241 tgtctccgtg tgtttgctgc tctggtttgc tcttcttcca gggccccttg gtaaaagaag
      301 agctgctgac agggtagagt ggaggtgggg aagtgaccac cagactggga gtgatgtagg
      361 ttgtggtcat caatgatgta ggttgctgtc accaggacac tcaaatcagc tgtgaccaga
      421 gccatctccc tctctgcctc ccgtttcccc tttactcttt actgttagtt ggcctggggt
      481 tcctgggctg aagtggagtt tcctcctgtc tccgtgtgtt tgctgctctg gtttgctctt
      541 cttccagggc cccttggtaa aagaagagct gctgacaggg tagagtggag gtggggaagt
      601 gaccaccaga ctgggagtga tgtaggttgt ggtcatcaat gatgtaggtt gctgtcacca
      661 ggacactcaa atcagctgtg accagagcca tctccctctc tgcctcccgt ttccccttta
      721 ctctttactg ttagttggcc tggggttcct gggctgaagt ggagtttcct cctgtctccg
      781 tgtgtttgct gctctggttt gctcttcttc cagggcccct tggtaaaaga agagctgctg
      841 acagggtaga gtggaggtgg ggaagtgacc accagactgg gagtgatgta ggttgtggtc
      901 atcaatgatg taggttgctg tcaccaggac actcaaatca gctgtgacca gaggagctcg
      961 ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc tgggatccag
     1021 atctttcgaa gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca
     1081 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1141 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ctgatctgca
     1201 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc tacggcgtgc
     1261 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1321 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1381 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1441 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1501 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     1561 acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1621 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc aagctgagca
     1681 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1741 tcactctcgg catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg ccgcgaattc
     1801 gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     1861 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     1921 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt cttgccacgg
     1981 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     2041 acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc atctgttgtt
     2101 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2161 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2221 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc acgtgcggac
     2281 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     2341 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     2401 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     2461 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     2521 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2581 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2641 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2701 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     2761 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2821 actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg
     2881 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2941 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     3001 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     3061 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3121 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     3181 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     3241 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3301 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3361 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3421 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3481 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3541 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     3601 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3661 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3721 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3781 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3841 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     3901 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3961 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4021 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4081 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4141 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4201 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     4261 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4321 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4381 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4441 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4501 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     4561 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4621 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4681 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4741 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4801 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4861 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4921 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4981 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
