LOCUS       sequence-432557-        5117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2420m 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 5117)
  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 5117)
  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: AiP1743 - pAAV-AiE2420m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(555..1346,1347..1415))
                     /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     1178..1197
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1416..1520)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1588..1606
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1644..1666)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1766..1785
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1802..2257)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1939..1960)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2151..2170
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2332..2472
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2343..2472
                     /label=AAV2 ITR
     polyA_signal    2590..2701
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2888..2908
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3001..3711,3712..3714,3715..3717))
                     /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(3043..3064)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3392..3411
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3651..3672
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3714..3723
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4722..4851
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4913..5117,1..384))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4993..5012)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
       61 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
      121 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
      181 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
      241 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
      301 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
      361 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
      421 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
      481 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
      541 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
      601 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
      661 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
      721 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
      781 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
      841 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
      901 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
      961 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     1021 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     1081 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     1141 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     1201 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     1261 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     1321 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     1381 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     1441 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     1501 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     1561 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     1621 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     1681 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     1741 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     1801 aaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     1861 ttttttaacc aatagaccga aatcggcaaa atcccttata aatcaaaaga atagcccgag
     1921 atagagttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     1981 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2041 aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2101 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2161 gcgaaaggag cgggcgctaa ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2221 acacccgccg cgcttaatgc gccgctacag ggcgcgtact atggttgctt tgacgtatgc
     2281 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc ccctgcaggc
     2341 agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct
     2401 ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca
     2461 ctaggggttc ctgcggccgc tcggtccgca cgtgagatct catgcctgct attgtcttcc
     2521 caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc
     2581 agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc
     2641 agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca
     2701 gtcgaagatc tctcgagcca cggaattgtc agtgcccaac agccgagccc ctgtccagca
     2761 gcgggcaagg caggcggcga tgagttccgc cgtggcaaga actaaccagg atttatacaa
     2821 ggaggagaaa atgaaagcca tacgggaagc aatagcatga tacaaaggca ttaaagcagc
     2881 gtatccacat agcgtaaaag gagcaacata gttaagaata ccagtcaatc tttcacaaat
     2941 tttgtaatcc agaggttgat tatgatatcg aattcgcggc cgcggcgcgc cgtcgactta
     3001 cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac
     3061 catgtgatcg cgcttctcgt tggggtcttt gctcagcttg gactggtagc tcaggtagtg
     3121 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
     3181 gagctgcacg ccgccgtcct cgatgttgtg gcggatcttg aagttggcct tgatgccgtt
     3241 cttctgcttg tcggcggtga tatagacgtt gtggctgttg tagttgtact ccagcttgtg
     3301 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
     3361 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa
     3421 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
     3481 catgtggtcg gggtagcggg cgaagcactg cacgccgtag cccagggtgg tcacgagggt
     3541 gggccagggc acgggcagct tgccggtggt gcagatcagc ttcagggtca gcttgccgta
     3601 ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc
     3661 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggt
     3721 ggcgaccggt agcgctagct tcgaaagatc tggatcccag aagcaaatgt aagcaataga
     3781 tggctctgcc ctgactttta tgcccagccc gagctccggt caaaggttat ttagtactaa
     3841 ccttgcttta tgagtagttg tattttaaat gattacatta atgtcatttg cactttaaaa
     3901 tgtgagacgc catcagtggc tatgtgttct aaatgttgac agactagaag agacaaataa
     3961 acctttaaca catattgtta aataagtata tgaagatgtt acataccatc caaccagagg
     4021 aagaagttat aagttaatgt gctgccattt cttctgtctc ccagacaata aagtgactat
     4081 agctgtggac cattagaaca cagaggcagt ctgagacaga cttcactttt ccctgacgac
     4141 tgccgttatc cactaaatac tttgttagag aattttctgc tttactctct agtggctgat
     4201 gatgcatcaa agaacacaga aatgtcctca cagcactaac attccaatgt tggagctggg
     4261 cagtaaatat aacataacca catttacaaa atgttaaatg gtgaaaagag tgcatttaaa
     4321 ataaacctga tgagctggag agatgactca gtggctgccc tttagaggac ccaggtttca
     4381 tttccaaaat ccatatggtg gcttaaaaac catccataag cccaggttcc ataacctcag
     4441 ccccccgaga ttcaatgggc acacacaggg tatggggggc aagatattca cacacgttca
     4501 gtaaaggtaa aataaatcca ataacctaat taactagcta gttagttgaa cctgagatgg
     4561 ccgttaagca atgaagtgag agtcagattg tacgttaggt aacgaggaga gtcctggctg
     4621 aggagatgag gttttaatga aagctgccaa ctagtataat atggattctc tgtagagacg
     4681 ataaaaaaca cgattattac tcttacaacg cgtgcggccg caggaacccc tagtgatgga
     4741 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     4801 ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gacatgtgag
     4861 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4921 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4981 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5041 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5101 tttctcatag ctcacgc
//
