LOCUS       sequence-466375-        4667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3282m 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 4667)
  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 4667)
  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: AiP20222 - pAAV-AiE3282m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   68..197
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(259..847)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(339..358)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1018..1809,1810..1878))
                     /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     1641..1660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1879..1983)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2051..2069
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2107..2129)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2229..2248
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2265..2720)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2402..2423)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2614..2633
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2795..2935
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2806..2935
                     /label=AAV2 ITR
     polyA_signal    3053..3164
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3351..3371
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3464..4174,4175..4177,4178..4180))
                     /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(3506..3527)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3855..3874
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4114..4135
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4177..4186
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4542..4553)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
ORIGIN
        1 atggggggca gggagggaac tgggcctccc caccccctgt caatatttat ataacgcgtg
       61 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
      121 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc
      181 gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      241 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      301 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      361 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      421 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      481 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      541 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      601 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      661 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
      721 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
      781 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      841 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
      901 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
      961 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1021 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1081 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1141 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1201 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1261 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1321 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1381 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1441 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1501 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1561 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1621 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1681 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1741 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1801 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1861 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     1921 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     1981 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     2041 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     2101 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     2161 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     2221 taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg taaacgttaa
     2281 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaatagac
     2341 cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagagt tgagtgttgt
     2401 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2461 aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg
     2521 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     2581 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     2641 taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     2701 tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac
     2761 agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg cgctcgctcg
     2821 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
     2881 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc
     2941 cgctcggtcc gcacgtgaga tctcatgcct gctattgtct tcccaatcct cccccttgct
     3001 gtcctgcccc accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat
     3061 ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac
     3121 gggggagggg caaacaacag atggctggca actagaaggc acagtcgaag atctctcgag
     3181 ccacggaatt gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg
     3241 cgatgagttc cgccgtggca agaactaacc aggatttata caaggaggag aaaatgaaag
     3301 ccatacggga agcaatagca tgatacaaag gcattaaagc agcgtatcca catagcgtaa
     3361 aaggagcaac atagttaaga ataccagtca atctttcaca aattttgtaa tccagaggtt
     3421 gattatgata tcgaattcgc ggccgcggcg cgccgtcgac ttacttgtac agctcgtcca
     3481 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     3541 cgttggggtc tttgctcagc ttggactggt agctcaggta gtggttgtcg ggcagcagca
     3601 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt
     3661 cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg
     3721 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     3781 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     3841 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     3901 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     3961 gggcgaagca ctgcacgccg tagcccaggg tggtcacgag ggtgggccag ggcacgggca
     4021 gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     4081 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     4141 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtagcgcta
     4201 gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat agatggctct gccctgactt
     4261 ttatgcccag cccgagctcc gccggcggcc acctgtcacc cggctaaggc accggaaggg
     4321 tattcagagc ctgtgcctct ccatcagcct tgctggacac ggagatgaac acagtccatg
     4381 ctagcgggag cgtagaacag ctcgagacaa gactgacccc accccaaccc tgagtcacga
     4441 tgctacccac atacccctcc aactcagtca ggatgtcatc catgtgccct cccagaacgc
     4501 ccaatcagaa tgctgcccac atgccttccc taaaacaaat cctgaagctc ccacctccct
     4561 tcctccctct accttcctcc agctaaacca gccaaagctg gtcggtatca gtaggcttgc
     4621 agctactgaa aaatctacct tctgttggcc aggcaacgaa ggagtct
//
