LOCUS       sequence-466236-        4754 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2068m 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 4754)
  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 4754)
  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: AiP1283 - pAAV-AiE2068m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     95..114
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   276..416
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   287..416
                     /label=AAV2 ITR
     polyA_signal    534..645
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     832..852
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(945..1655,1656..1658,1659..1661))
                     /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(987..1008)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1336..1355
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1595..1616
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1658..1667
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2303..2432
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2494..3082)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2574..2593)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3253..4044,4045..4113))
                     /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     3876..3895
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4114..4218)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4286..4304
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4342..4364)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4464..4483
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4500..4754,1..201))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4637..4658)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
       61 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
      121 ctaaggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
      181 atgcgccgct acagggcgcg tactatggtt gctttgacgt atgcggtgtg aaataccgca
      241 cagatgcgta aggagaaaat accgcatcag gcgcccctgc aggcagctgc gcgctcgctc
      301 gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc
      361 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg
      421 ccgctcggtc cgcacgtgag atctcatgcc tgctattgtc ttcccaatcc tcccccttgc
      481 tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa
      541 tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca
      601 cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgaa gatctctcga
      661 gccacggaat tgtcagtgcc caacagccga gcccctgtcc agcagcgggc aaggcaggcg
      721 gcgatgagtt ccgccgtggc aagaactaac caggatttat acaaggagga gaaaatgaaa
      781 gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc acatagcgta
      841 aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta atccagaggt
      901 tgattatgat atcgaattcg cggccgcggc gcgccgtcga cttacttgta cagctcgtcc
      961 atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc
     1021 tcgttggggt ctttgctcag cttggactgg tagctcaggt agtggttgtc gggcagcagc
     1081 acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgccgccg
     1141 tcctcgatgt tgtggcggat cttgaagttg gccttgatgc cgttcttctg cttgtcggcg
     1201 gtgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg
     1261 tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac
     1321 ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg
     1381 acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag
     1441 cgggcgaagc actgcacgcc gtagcccagg gtggtcacga gggtgggcca gggcacgggc
     1501 agcttgccgg tggtgcagat cagcttcagg gtcagcttgc cgtaggtggc atcgccctcg
     1561 ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg
     1621 ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tggtggcgac cggtagcgct
     1681 agcttcgaaa gatctggatc ccagaagcaa atgtaagcaa tagatggctc tgccctgact
     1741 tttatgccca gcccgagctc ggtctgaaca aacatgcttg gcttttgttc aagtagtttt
     1801 tcagtcattt gttggcttaa aaaatttttt tacagagtct caaggagcat acaaggtgct
     1861 gaactatctg ggggtggggc atctggcccc attccacaag gaactgaagt tctttagtgg
     1921 ccaggcacat tggatgacag cacaggcccc agaatcaggt ctggatgaag tcttggctca
     1981 tcaagacatt cctaacatgg ccttagctgg gtcatttctc ctctcaaatc atcagtttct
     2041 tttttagtaa attgagagtc ataaatctga actcatagta aggactgggt cagaccatga
     2101 acagaagcat ttgatctgac gttgctaagt tctcaataat accaggtgcc gttagcaacc
     2161 cctgagcgtg gtctgaggag gcgagagccc tgcacactga gcagcttgta gaggcctgag
     2221 ggatggttgt gcatttggtc ttctgaaagg atcttgttga ctaaagctgc ttctttgttt
     2281 cttcctggac gcgtgcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     2341 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggcggcctc
     2401 agtgagcgag cgagcgcgca gctgcctgca ggacatgtga gcaaaaggcc agcaaaaggc
     2461 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2521 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2581 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2641 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2701 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2761 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2821 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2881 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2941 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3001 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3061 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3121 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3181 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3241 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3301 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3361 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3421 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3481 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3541 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3601 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3661 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3721 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3781 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3841 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3901 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3961 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4021 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4081 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4141 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4201 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     4261 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     4321 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     4381 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     4441 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa
     4501 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     4561 ttttaaccaa tagaccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat
     4621 agagttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     4681 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa
     4741 atcaagtttt ttgg
//
