LOCUS       sequence-466254-        5239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2166m 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 5239)
  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 5239)
  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: AiP1395 - pAAV-AiE2166m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(66..654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(146..165)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(825..1616,1617..1685))
                     /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     1448..1467
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1686..1790)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1858..1876
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1914..1936)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2036..2055
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2072..2527)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2209..2230)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2421..2440
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2602..2742
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2613..2742
                     /label=AAV2 ITR
     polyA_signal    2860..2971
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3158..3178
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3271..3981,3982..3984,3985..3987))
                     /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(3313..3334)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3662..3681
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3921..3942
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3984..3993
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   join(5114..5239,1..4)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 caggacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
       61 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      121 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
      181 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
      241 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
      301 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
      361 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
      421 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
      481 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
      541 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
      601 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
      661 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
      721 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
      781 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
      841 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
      901 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
      961 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     1021 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     1081 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     1141 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     1201 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     1261 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     1321 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     1381 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     1441 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     1501 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     1561 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     1621 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     1681 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     1741 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     1801 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     1861 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     1921 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     1981 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     2041 tcagagcaga ttgtactgag agtgcaccat aaaattgtaa acgttaatat tttgttaaaa
     2101 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aatagaccga aatcggcaaa
     2161 atcccttata aatcaaaaga atagcccgag atagagttga gtgttgttcc agtttggaac
     2221 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     2281 ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc gaggtgccgt
     2341 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     2401 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctaa ggcgctggca
     2461 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     2521 ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat accgcacaga tgcgtaagga
     2581 gaaaataccg catcaggcgc ccctgcaggc agctgcgcgc tcgctcgctc actgaggccg
     2641 cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
     2701 cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc tcggtccgca
     2761 cgtgagatct catgcctgct attgtcttcc caatcctccc ccttgctgtc ctgccccacc
     2821 ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat
     2881 taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa
     2941 acaacagatg gctggcaact agaaggcaca gtcgaagatc tctcgagcca cggaattgtc
     3001 agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc
     3061 cgtggcaaga actaaccagg atttatacaa ggaggagaaa atgaaagcca tacgggaagc
     3121 aatagcatga tacaaaggca ttaaagcagc gtatccacat agcgtaaaag gagcaacata
     3181 gttaagaata ccagtcaatc tttcacaaat tttgtaatcc agaggttgat tatgatatcg
     3241 aattcgcggc cgcggcgcgc cgtcgactta cttgtacagc tcgtccatgc cgagagtgat
     3301 cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt
     3361 gctcagcttg gactggtagc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc
     3421 gatgggggtg ttctgctggt agtggtcggc gagctgcacg ccgccgtcct cgatgttgtg
     3481 gcggatcttg aagttggcct tgatgccgtt cttctgcttg tcggcggtga tatagacgtt
     3541 gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc
     3601 gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg
     3661 ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg
     3721 catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg cgaagcactg
     3781 cacgccgtag cccagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt
     3841 gcagatcagc ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac
     3901 gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt
     3961 gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt agcgctagct tcgaaagatc
     4021 tggatcccag aagcaaatgt aagcaataga tggctctgcc ctgactttta tgcccagccc
     4081 gagctcctgc atgcccaggg ttaccaggat accacacgac ataccctctc acccccactt
     4141 ccttcctctt catccctgga gagcctaata cctgtccaca cccagaaaac ttcggtgagc
     4201 ctccctggtt ttcccctcag ttctgggatt tcatggctta tgactggttt gtggtacatt
     4261 ccacactgtt ttcttcatcc accctcttcc agcacagagc acttcctcaa gacagacagc
     4321 cttctccaca actgggagca gcgtagcttg gggcacagaa cacatcaaac catgcatcat
     4381 cggactcgag ctttgagcct gggatcttta agtttctcct aaacctaaag atattgtgga
     4441 accttgtggc tctggggtag ggagtaggct gccctttggg aggcacgtgg atacccataa
     4501 caatctccta tggttctgat acagctcacc ctgtgctagg tctcacacac accatacaca
     4561 tatatacatg acacgtgtat atgtatacat acacacaaac acatactagg aacactacac
     4621 ataacacaca tatgtacata cttacgtaat acatatacat atacacacac atacacatac
     4681 cagacacaca ccttcacact gtctgtcctt acaacatact catatatgcc ctggataatc
     4741 acaggctgca ttgcgggtgt ctggatgcct ttgaaaggtc caacagtgac tgctgtgcag
     4801 cgtccatgag cttgggacat ggcataatca gggctaatga cacagcactc actgtgcgat
     4861 tgcctcgctg ctatgctgta atcagcatgt aattaagtgg tcaccagtca ccacgtaaac
     4921 acggcttata ctagatgcat ctgtagctta aatagcccca caaagtcact ttaatttcaa
     4981 tgttgaatgg aaaacagtca gttggcatgt gtggagaaca cggggttgga aatgtgaggc
     5041 ccctcaggaa ggaggggggc atcatctcca gcctcagttg caaagcagga acgggccaga
     5101 cgcgtgcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     5161 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcggcct cagtgagcga
     5221 gcgagcgcgc agctgcctg
//
