LOCUS       sequence-466371-        4711 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2637m 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 4711)
  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 4711)
  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: AiP20069 - pAAV-AiE2637m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   90..219
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(281..869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(361..380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1040..1831,1832..1900))
                     /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     1663..1682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1901..2005)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2073..2091
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2129..2151)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2251..2270
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2287..2742)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2424..2445)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2636..2655
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2817..2957
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2828..2957
                     /label=AAV2 ITR
     polyA_signal    3075..3186
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3373..3393
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3486..4196,4197..4199,4200..4202))
                     /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(3528..3549)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3877..3896
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4136..4157
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4199..4208
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gattttccac cggctgcaaa ctggtctgcc atagatgttc caaagttgtg cttcgggggt
       61 tggggtgggg ggtgaacgcg tgcggccgca ggaaccccta gtgatggagt tggccactcc
      121 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
      181 cggcctcagt gagcgagcga gcgcgcagct gcctgcagga catgtgagca aaaggccagc
      241 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
      301 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
      361 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
      421 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
      481 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
      541 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
      601 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
      661 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
      721 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
      781 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
      841 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
      901 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
      961 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     1021 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     1081 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     1141 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     1201 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     1261 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     1321 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     1381 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     1441 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     1501 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     1561 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     1621 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     1681 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     1741 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     1801 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     1861 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     1921 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     1981 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     2041 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     2101 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     2161 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     2221 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     2281 accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2341 gctcattttt taaccaatag accgaaatcg gcaaaatccc ttataaatca aaagaatagc
     2401 ccgagataga gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2461 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2521 cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2581 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2641 agaaagcgaa aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2701 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt tgctttgacg
     2761 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgcccctg
     2821 caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc
     2881 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
     2941 catcactagg ggttcctgcg gccgctcggt ccgcacgtga gatctcatgc ctgctattgt
     3001 cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc
     3061 tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac
     3121 cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag
     3181 gcacagtcga agatctctcg agccacggaa ttgtcagtgc ccaacagccg agcccctgtc
     3241 cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caagaactaa ccaggattta
     3301 tacaaggagg agaaaatgaa agccatacgg gaagcaatag catgatacaa aggcattaaa
     3361 gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt caatctttca
     3421 caaattttgt aatccagagg ttgattatga tatcgaattc gcggccgcgg cgcgccgtcg
     3481 acttacttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc
     3541 aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gcttggactg gtagctcagg
     3601 tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg
     3661 tcggcgagct gcacgccgcc gtcctcgatg ttgtggcgga tcttgaagtt ggccttgatg
     3721 ccgttcttct gcttgtcggc ggtgatatag acgttgtggc tgttgtagtt gtactccagc
     3781 ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc
     3841 accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg
     3901 aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc
     3961 tgcttcatgt ggtcggggta gcgggcgaag cactgcacgc cgtagcccag ggtggtcacg
     4021 agggtgggcc agggcacggg cagcttgccg gtggtgcaga tcagcttcag ggtcagcttg
     4081 ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg
     4141 ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc
     4201 atggtggcga ccggtagcgc tagcttcgaa agatctggat cccagaagca aatgtaagca
     4261 atagatggct ctgccctgac ttttatgccc agcccgagct ccgccggcga atttgcatat
     4321 ttttgtccct ctctttgtct gcatggaggg tgctaactca accaagaatt ctgttcacat
     4381 ggtagtcatt tgctctgctt agatggtcca catagcctta ccccggccaa agatatattt
     4441 gccagagtct taatgtagtg ataaatttta tctcttggtc tttcttttat gtaacatttc
     4501 ctagaaatgc caaaatgtgg gtgagggagg ctgagtatca agttcggagc attgctaact
     4561 ttagacagtg taaagtcccc acgccacgct gcagactttc actctttatt tcagacccag
     4621 gctgaagttg ctccccccat cctccacctc ctcttgtgcc accacaaatg aataccatct
     4681 gatggctgga gaaatttaac gcgaatccag t
//
