LOCUS       sequence-466328-        4837 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2385m 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 4837)
  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 4837)
  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: AiP1913 - pAAV-AiE2385m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4837
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   153..282
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(344..932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(424..443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1103..1894,1895..1963))
                     /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     1726..1745
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1964..2068)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2136..2154
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2192..2214)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2314..2333
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2350..2805)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2487..2508)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2699..2718
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2880..3020
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2891..3020
                     /label=AAV2 ITR
     polyA_signal    3138..3249
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3436..3456
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3549..4259,4260..4262,4263..4265))
                     /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(3591..3612)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3940..3959
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4199..4220
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4262..4271
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tgagcaggac ctgtgtaagg cattggaggg ttgaaaggaa ttatcagatt tgttttgaac
       61 tgcttctaga atttttttaa agctaattgg tttaagggcg tctccactca ggccagagag
      121 gagacaagta atgaaatggt ctctgcggcc gcaggaaccc ctagtgatgg agttggccac
      181 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
      241 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggacatgtga gcaaaaggcc
      301 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
      361 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
      421 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
      481 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
      541 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
      601 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
      661 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
      721 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
      781 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
      841 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
      901 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
      961 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     1021 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     1081 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     1141 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     1201 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     1261 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     1321 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     1381 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     1441 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     1501 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     1561 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     1621 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     1681 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     1741 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     1801 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     1861 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     1921 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     1981 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     2041 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     2101 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     2161 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     2221 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     2281 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     2341 tgcaccataa aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2401 tcagctcatt ttttaaccaa tagaccgaaa tcggcaaaat cccttataaa tcaaaagaat
     2461 agcccgagat agagttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     2521 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     2581 catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     2641 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     2701 ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag tgtagcggtc acgctgcgcg
     2761 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtactat ggttgctttg
     2821 acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgccc
     2881 ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg
     2941 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
     3001 ctccatcact aggggttcct gcggccgctc ggtccgcacg tgagatctca tgcctgctat
     3061 tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga atagaatgac
     3121 acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg
     3181 caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag
     3241 aaggcacagt cgaagatctc tcgagccacg gaattgtcag tgcccaacag ccgagcccct
     3301 gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaagaac taaccaggat
     3361 ttatacaagg aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt
     3421 aaagcagcgt atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt
     3481 tcacaaattt tgtaatccag aggttgatta tgatatcgaa ttcgcggccg cggcgcgccg
     3541 tcgacttact tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
     3601 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga ctggtagctc
     3661 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
     3721 tggtcggcga gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa gttggccttg
     3781 atgccgttct tctgcttgtc ggcggtgata tagacgttgt ggctgttgta gttgtactcc
     3841 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg
     3901 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc
     3961 ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg
     4021 tgctgcttca tgtggtcggg gtagcgggcg aagcactgca cgccgtagcc cagggtggtc
     4081 acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatcagctt cagggtcagc
     4141 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg
     4201 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc
     4261 accatggtgg cgaccggtag cgctagcttc gaaagatctg gatcccagaa gcaaatgtaa
     4321 gcaatagatg gctctgccct gacttttatg cccagcccga gctccgagca ggaatcaaag
     4381 cagaaccttc attttagttg gtcctcagaa aggcaggtta gcttagagac attttttttc
     4441 caccatcctg aattaattgg tgtagtggtt acgttctctg aaaaaccgac aagctctcct
     4501 tttctcgctg ttaagagatc taaggcctga cagttttgga gagggcccca ggccagagaa
     4561 gtaagctgat gttgggcaga agccagagat tgaaccatgg agtctacaga gagctgtaat
     4621 ttggtcacaa aatcatgagc tgtgaggacg gaatgtccca aagcacttct ggccatgact
     4681 atcccagtta aggcagtagc cagacttgtg ccaataatta taggcagcga tgcagccttg
     4741 ggggatgagc tttttagaag agccagtagc attcagattg gacataactc tcatttgggg
     4801 actattgcca ctaaaaaact ggatccagtt gcatatt
//
