LOCUS       sequence-461925-        4682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0086h_3xC2 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 4682)
  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 4682)
  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: AiP13901 -
            pAAV-AiE0086h_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4682
                     /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      694..703
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(700..702,703..705,706..1416)
                     /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(745..766)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1006..1025)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1353..1374
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1509..1529)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1716..1827
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1945..2085
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1945..2074
                     /label=AAV2 ITR
     rep_origin      2160..2615
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2247..2266)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2457..2478
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2632..2651)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2751..2773
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2811..2829)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2897..3001
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3002..3070,3071..3862)
                     /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(3220..3239)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4033..4621
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4522..4541
                     /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 gcgtgaggct gctgcataac aatctccttc cttgattaga
      181 gcatcttaat tatgctccag gaatttgctt aaacagatta atcattctgg aggacagatt
      241 tagaagctgt gctagctgag ggatctgctc tgagtgtccc cggggagatt ctggtggagg
      301 ctgctgcata acaatctcct tccttgatta gagcatctta attatgctcc aggaatttgc
      361 ttaaacagat taatcattct ggaggacaga tttagaagct gtgctagctg agggatctgc
      421 tctgagtgtc cccggggaga ttctggtgga ggctgctgca taacaatctc cttccttgat
      481 tagagcatct taattatgct ccaggaattt gcttaaacag attaatcatt ctggaggaca
      541 gatttagaag ctgtgctagc tgagggatct gctctgagtg tccccgggga gattctggtg
      601 gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg
      661 gatccagatc tttcgaagct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag
      721 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      781 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg
      841 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac
      901 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
      961 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1021 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1081 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1141 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     1201 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     1261 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag
     1321 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1381 gccgggatca ctctcggcat ggacgagctg tacaagtaag tcgacggcgc gccgcggccg
     1441 cgaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     1501 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     1561 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt
     1621 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     1681 ggcactgaca attccgtggc tcgagagatc ttcgactgtg ccttctagtt gccagccatc
     1741 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     1801 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     1861 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat
     1921 ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     1981 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     2041 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     2101 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     2161 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2221 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2281 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2341 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     2401 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2461 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     2521 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2581 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     2641 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     2701 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     2761 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     2821 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     2881 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2941 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3001 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     3061 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     3121 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3181 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3241 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3301 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3361 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3421 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3481 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3541 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3601 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3661 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3721 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3781 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3841 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3901 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3961 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4021 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4081 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4141 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4201 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4261 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4321 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4381 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4441 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4501 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4561 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4621 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4681 gt
//
