LOCUS       sequence-469031-        4971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ExE0581m 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 4971)
  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 4971)
  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: AiP12730 - pAAV-ExE0581m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      983..992
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(989..991,992..994,995..1705)
                     /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(1034..1055)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1295..1314)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1642..1663
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1798..1818)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2005..2116
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2234..2374
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2234..2363
                     /label=AAV2 ITR
     rep_origin      2449..2904
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2536..2555)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2746..2767
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2921..2940)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3040..3062
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3100..3118)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3186..3290
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3291..3359,3360..4151)
                     /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     complement(3509..3528)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4322..4910
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4811..4830
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgctgtg ggtataggag gcatgtaaaa ttaatataat
      181 gttttgatat ttaggagaaa attgcaaaac aaaatgatga cagtgtttga aagtgtttga
      241 tcagtgccaa gcatcacttt atgtacttgg caaacatgac ttgaggcctt aagctgtgat
      301 ttgcaaatgt agattggaat caagatcttt atagatgagg aagcaaaaat cagaagacaa
      361 aataacatta tcaacttgat ctcatgtgca gccagggctg aactgcaaat gctgatttgc
      421 cccagtctgg gctcctcaaa tcgttccttg gaatcctatt agttggaact ttatctctgc
      481 tcgtggcagg gtgcctggga ccatgtttat aaatatctgc tgaatgaaga ataagtgagt
      541 caatcgaacc agaactcact ttggttagtt aatttcattc gtggtattta tggagagcag
      601 aagaaagaat tccagagaca cgatttgtca aaactctcta aagaaaatga tgacactata
      661 tattgatgaa aatgaatgtt cttgttcttg ctttatttga ttttcttgtc cccccactcc
      721 ccatctgcta gggtctcatt acagcatagt tcttgaatat cccaggttga cctgaagtta
      781 caatatattc ttgatttaga tggcagacat tgggaatatt ttgactctta aaatttaata
      841 gctaatcttt gaagaaaatt aaaatagtga tctccatttt gtagacaagg agctcgggct
      901 gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct
      961 ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     1021 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1081 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac
     1141 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg
     1201 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1261 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1321 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1381 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1441 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     1501 catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1561 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga
     1621 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1681 tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc gaattcgata
     1741 tcataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     1801 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     1861 ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga
     1921 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     1981 ttccgtggct cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2041 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2101 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2161 ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg
     2221 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     2281 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     2341 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     2401 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     2461 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2521 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2581 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2641 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     2701 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2761 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     2821 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     2881 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     2941 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3001 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3061 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     3121 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     3181 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3241 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3301 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3361 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3421 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3481 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3541 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3601 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3661 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3721 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3781 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3841 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3901 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3961 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4021 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4081 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4141 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4201 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4261 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4321 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4381 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4441 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4501 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4561 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4621 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4681 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4741 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4801 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4861 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4921 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
