LOCUS       sequence-466299-        4536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2418m 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 4536)
  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 4536)
  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: AiP1750 - pAAV-AiE2418m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   164..304
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   175..304
                     /label=AAV2 ITR
     polyA_signal    422..533
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     720..740
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(833..1543,1544..1546,1547..1549))
                     /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(875..896)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1224..1243
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1483..1504
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1546..1555
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   1973..2102
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2164..2752)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2244..2263)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2923..3714,3715..3783))
                     /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     3546..3565
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3784..3888)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3956..3974
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4012..4034)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4134..4153
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4170..4536,1..89))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4307..4328)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(4519..4536,1..2)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
       61 cgcgcttaat gcgccgctac agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa
      121 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc
      181 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc
      241 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
      301 tcctgcggcc gctcggtccg cacgtgagat ctcatgcctg ctattgtctt cccaatcctc
      361 ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg
      421 cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa
      481 ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaaga
      541 tctctcgagc cacggaattg tcagtgccca acagccgagc ccctgtccag cagcgggcaa
      601 ggcaggcggc gatgagttcc gccgtggcaa gaactaacca ggatttatac aaggaggaga
      661 aaatgaaagc catacgggaa gcaatagcat gatacaaagg cattaaagca gcgtatccac
      721 atagcgtaaa aggagcaaca tagttaagaa taccagtcaa tctttcacaa attttgtaat
      781 ccagaggttg attatgatat cgaattcgcg gccgcggcgc gccgtcgact tacttgtaca
      841 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
      901 cgcgcttctc gttggggtct ttgctcagct tggactggta gctcaggtag tggttgtcgg
      961 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
     1021 cgccgccgtc ctcgatgttg tggcggatct tgaagttggc cttgatgccg ttcttctgct
     1081 tgtcggcggt gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga
     1141 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
     1201 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc
     1261 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
     1321 cggggtagcg ggcgaagcac tgcacgccgt agcccagggt ggtcacgagg gtgggccagg
     1381 gcacgggcag cttgccggtg gtgcagatca gcttcagggt cagcttgccg taggtggcat
     1441 cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     1501 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg
     1561 gtagcgctag cttcgaaaga tctggatccc agaagcaaat gtaagcaata gatggctctg
     1621 ccctgacttt tatgcccagc ccgagctccg aggggaggtg aaggcaaccg gagtttcatg
     1681 ggaccgaggg gtgggcacga gcgagcagca gctgaggaag ccaccggcat cccatgccac
     1741 tgccttatcc cacttgggta cgcgatggat gatgctgagc aggttttagg gacagatatt
     1801 aatatagatg gagaagagga aggcttcgag cccctgagaa cacactacat aatgcactga
     1861 aaatatccat cctagaaaca gaacgcagag gaccgctgtg aggcggggcg cgcaggtgca
     1921 gagagcaggg gcctcagtca tcgattatta ctcttacaac gcgtgcggcc gcaggaaccc
     1981 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     2041 ccaaaggtcg cccgacgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     2101 ggacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2161 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2221 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2281 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2341 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2401 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2461 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2521 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2581 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2641 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2701 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2761 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2821 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2881 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2941 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3001 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3061 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3121 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3181 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3241 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3301 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3361 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3421 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3481 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3541 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3601 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3661 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3721 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3781 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3841 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3901 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     3961 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     4021 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     4081 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     4141 agagcagatt gtactgagag tgcaccataa aattgtaaac gttaatattt tgttaaaatt
     4201 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa tcggcaaaat
     4261 cccttataaa tcaaaagaat agcccgagat agagttgagt gttgttccag tttggaacaa
     4321 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4381 cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa
     4441 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4501 gaacgtggcg agaaaggaag ggaagaaagc gaaagg
//
