LOCUS       sequence-466373-        5173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3287m 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 5173)
  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 5173)
  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: AiP20128 - pAAV-AiE3287m-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      481..490
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(487..489,490..492,493..1203)
                     /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(532..553)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(793..812)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1140..1161
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1296..1316)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1503..1614
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1732..1872
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1732..1861
                     /label=AAV2 ITR
     rep_origin      1947..2402
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2034..2053)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2244..2265
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2419..2438)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2538..2560
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2598..2616)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2684..2788
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2789..2857,2858..3649)
                     /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(3007..3026)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3820..4408
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4309..4328
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4470..4599
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 caacatttca ccacttgcat ttctatactg gcaggtggcg tcttgcttcc aacttctcag
       61 aaccccaact cagcataaca gaaaggcaag ggggctcagg gcggtagagg aagcctgctc
      121 ccgtgggagg tacactattt accagcccga tttaaacccg ctacggaggg ggcacggtcc
      181 tggaagggcg gagcagccct agtgggcggg gcgggggcgg ggctccgagg gactaagctg
      241 ggtacaggaa agaccggagc tctcagagcc acgtctgtct ctttaaatgc cacgggctgc
      301 actcccgccc gggagccccg agccggatct acaatcccgt ccctcccgct ccggcgcttg
      361 ttgctcgcct agcccgagcg ggagctgcgg cccccgggaa acccagctag ctccttcggc
      421 ttccctcctc gtgccccacc ttcctggaaa tctgcgtgag gggtgctagc gctaccggtc
      481 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      541 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      601 acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg
      661 cccaccctcg tgaccaccct gggctacggc gtgcagtgct tcgcccgcta ccccgaccac
      721 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
      781 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
      841 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
      901 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag
      961 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     1021 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1081 aaccactacc tgagctacca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     1141 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1201 aagtaagtcg acggcgcgcc gcggccgcga attcgatatc ataatcaacc tctggattac
     1261 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     1321 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     1381 tccttgtata aatcctggtt agttcttgcc acggcggaac tcatcgccgc ctgccttgcc
     1441 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggctcg agagatcttc
     1501 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     1561 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     1621 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     1681 ttgggaagac aatagcaggc atgagatctc acgtgcggac cgagcggccg caggaacccc
     1741 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     1801 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     1861 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     1921 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     1981 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     2041 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2101 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2161 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2221 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     2281 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     2341 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     2401 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     2461 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     2521 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     2581 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     2641 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2701 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2761 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2821 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2881 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2941 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3001 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3061 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3121 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3181 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3241 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3301 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3361 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3421 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3481 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3541 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3601 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3661 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3721 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3781 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3841 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3901 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3961 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4021 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4081 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4141 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4201 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4261 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4321 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4381 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4441 ggccttttgc tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac
     4501 tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
     4561 gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgcacg cgttgtaaga
     4621 gtaataatcg ccggcgaaat cactttagta gtttaggtct ggggtagaga aatgacccca
     4681 gcagggaaga aagtgtcctc ttgaggaggt taataaccaa caaggctgtc acttccctca
     4741 ggaggaagga aatatattca ccaggagtct ctgctaattt gtttttaatt taaggacaat
     4801 taagcaactc tacaccccca tcccatttct gcatgctttc tctcacaact agaaaagaaa
     4861 tctaaaacaa agaaggctgt gaagttaatg ttgagtttac ttggagggac acctaacagt
     4921 cattagggat ttgaggagct agtcaactaa gtcctgtatt atactgcttc ctctttctga
     4981 attattatta ttattattat ttactatcca aaattattac ttgctctgga aacaggtttt
     5041 agctgatgtg tttcttgaac tgttgcagca cagactactt taggttataa aaacgttctg
     5101 ataagaaaag tctgtctctc tttctttaga tactccatct aagaagaaat ggaaggagaa
     5161 gaggaagaaa gac
//
