LOCUS       sequence-432581-        4797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2585m 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 4797)
  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 4797)
  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: AiP1970 - pAAV-AiE2585m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   73..213
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   84..213
                     /label=AAV2 ITR
     polyA_signal    331..442
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     629..649
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(742..1452,1453..1455,1456..1458))
                     /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(784..805)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1133..1152
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1392..1413
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1455..1464
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2143..2272
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2334..2922)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2414..2433)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3093..3884,3885..3953))
                     /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     3716..3735
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3954..4058)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4126..4144
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4182..4204)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4304..4323
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4340..4795)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4477..4498)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4689..4708
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tatggttgct ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
       61 gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
      121 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
      181 agggagtggc caactccatc actaggggtt cctgcggccg ctcggtccgc acgtgagatc
      241 tcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
      301 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
      361 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
      421 ggctggcaac tagaaggcac agtcgaagat ctctcgagcc acggaattgt cagtgcccaa
      481 cagccgagcc cctgtccagc agcgggcaag gcaggcggcg atgagttccg ccgtggcaag
      541 aactaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag caatagcatg
      601 atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat agttaagaat
      661 accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatgatatc gaattcgcgg
      721 ccgcggcgcg ccgtcgactt acttgtacag ctcgtccatg ccgagagtga tcccggcggc
      781 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt
      841 ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
      901 gttctgctgg tagtggtcgg cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt
      961 gaagttggcc ttgatgccgt tcttctgctt gtcggcggtg atatagacgt tgtggctgtt
     1021 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     1081 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     1141 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     1201 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact gcacgccgta
     1261 gcccagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatcag
     1321 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     1381 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     1441 ctcgcccttg ctcaccatgg tggcgaccgg tagcgctagc ttcgaaagat ctggatccca
     1501 gaagcaaatg taagcaatag atggctctgc cctgactttt atgcccagcc cgagctccgt
     1561 aactaggtct acatgaggac atgagggtgg aggcccctgc aaatctgtgt ccttgggaga
     1621 ccctagagag cttgctttct atctctgtgt gccacctgag cacccagaga gaaagcagtt
     1681 atctgaacgt taggaagaga atctttatca gaaaccagtc aagccaaaga cccaatctgg
     1741 gacttgccag tcttctaaac tgtgaaagaa tacatttcca ctgctctggc cactcattct
     1801 ctgatattta attatagcag ccctagccaa ctaacactga ctgagagagg aattctttcc
     1861 agggagagat gcaaattatt tctgaaagta caaaagatga ctccagagct aatggctttt
     1921 tgccctgcaa tgaattaata tgtactgcat gtaattaaca atcttatttc tgccatttcc
     1981 ttttgatagc tctatatgtg ctatcttatt gcttgaacta tcatcatagt tttaatgagc
     2041 tgaaataaaa taattgacac atactcacca tgcatttgta ccaaaattat ggggttttat
     2101 atattttaaa ataatacttc aaacagagag agatgcggcc gcaggaaccc ctagtgatgg
     2161 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     2221 cccgacgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggacatgtga
     2281 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2341 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2401 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2461 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2521 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2581 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2641 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2701 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2761 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2821 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2881 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2941 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3001 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3061 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3121 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3181 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3241 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3301 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3361 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3421 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3481 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3541 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3601 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3661 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3721 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3781 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3841 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3901 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3961 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4021 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4081 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     4141 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     4201 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     4261 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     4321 gtactgagag tgcaccataa aattgtaaac gttaatattt tgttaaaatt cgcgttaaat
     4381 ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa tcggcaaaat cccttataaa
     4441 tcaaaagaat agcccgagat agagttgagt gttgttccag tttggaacaa gagtccacta
     4501 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4561 ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4621 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4681 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag tgtagcggtc
     4741 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtac
//
