LOCUS       sequence-470805-        4871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0359h_3xC1 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 4871)
  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 4871)
  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: AiP12561 -
            pAAV-AiE0359h_3xC1-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4871
                     /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      883..892
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(889..891,892..894,895..1605)
                     /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(934..955)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1195..1214)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1542..1563
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1698..1718)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1905..2016
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2134..2274
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2134..2263
                     /label=AAV2 ITR
     rep_origin      2349..2804
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2436..2455)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2646..2667
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2821..2840)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2940..2962
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3000..3018)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3086..3190
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3191..3259,3260..4051)
                     /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(3409..3428)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4222..4810
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4711..4730
                     /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 gcggccgcac gcgtatcaaa gaatcaggtg catattttca cagctccttg
      181 gtctgaactg gtacccagga tcaagatgtc cctcccatga ttctaatctc cagccccctt
      241 acaccatagt cacttccaaa ctctggattt cttaacattg cagtttgctt ccaaaaaaaa
      301 attaaaaaaa aaacctcatc tgggcattag aagtcattct gaagaggctg tcctatatca
      361 tcaaagaatc aggtgcatat tttcacagct ccttggtctg aactggtacc caggatcaag
      421 atgtccctcc catgattcta atctccagcc cccttacacc atagtcactt ccaaactctg
      481 gatttcttaa cattgcagtt tgcttccaaa aaaaaattaa aaaaaaaacc tcatctgggc
      541 attagaagtc attctgaaga ggctgtccta tatcatcaaa gaatcaggtg catattttca
      601 cagctccttg gtctgaactg gtacccagga tcaagatgtc cctcccatga ttctaatctc
      661 cagccccctt acaccatagt cacttccaaa ctctggattt cttaacattg cagtttgctt
      721 ccaaaaaaaa attaaaaaaa aaacctcatc tgggcattag aagtcattct gaagaggctg
      781 tcctatatcg agctcgggct gggcataaaa gtcagggcag agccatctat tgcttacatt
      841 tgcttctggg atccagatct ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag
      901 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      961 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1021 ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1081 ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     1141 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1201 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1261 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1321 aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc
     1381 aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag
     1441 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
     1501 cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1561 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagt cgacggcgcg
     1621 ccgcggccgc gaattcgata tcataatcaa cctctggatt acaaaatttg tgaaagattg
     1681 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     1741 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     1801 ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     1861 cggctgttgg gcactgacaa ttccgtggct cgagagatct tcgactgtgc cttctagttg
     1921 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     1981 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2041 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     2101 gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     2161 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     2221 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     2281 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     2341 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2401 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     2461 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2521 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     2581 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2641 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     2701 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     2761 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     2821 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     2881 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     2941 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     3001 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     3061 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3121 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3181 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3241 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3301 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3361 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3421 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3481 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3541 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3601 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3661 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3721 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3781 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3841 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3901 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3961 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4021 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4081 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4141 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4201 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4261 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4321 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4381 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4441 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4501 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4561 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4621 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4681 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4741 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4801 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4861 tgctcacatg t
//
