LOCUS       sequence-473482-        4715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0025m 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 4715)
  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 4715)
  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: AiP11282 - scAAV-AiE0025m-minBG-SYFP2-WPRE3-BG...
FEATURES             Location/Qualifiers
     source          1..4715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      705..714
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(711..713,714..716,717..1427)
                     /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(756..777)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1017..1036)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1364..1385
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1520..1540)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1720..1831
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2070..2087)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2070..2086)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2079..2101)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2295..2314)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2414..2436
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2474..2492)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2560..2664
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2665..2733,2734..3525)
                     /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(2883..2902)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3696..4284
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4185..4204
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4572..4593
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4608..4613,4614..4631,4632..4638)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4646..4662
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4651..4673
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4670..4686
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4670..4686
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaagcttccc ggggggatct gggccactcc ctctctgcgc gctcgctcgc tcactgaggc
       61 cgcccgggca aagcccgggc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      121 aactccatca ctaggggttc ctggaggggt ggagtcgtga cctaggacgc gtgctccaac
      181 atttgcagtc atataataga tgtttctttt agagttattt cagttcattt caataagaaa
      241 cagtagctaa gggaaaaatt catggcatta caattgatat taaaagaacc aatgagcttt
      301 gccactaatc tgagcacttt tatttacttc agagaatgat accgattatt aagcacatac
      361 tttcttccat ttttgttgtt cctgattggt gccagggttt tcttggcttg ttgcgggata
      421 ggctgaatag accaactttc ctttgcatct gcatattcga taggtctagt gattctgaat
      481 ccaagtcact tgaataatga tggaaccatt ttgttcctga tgaatacaca ctaatataat
      541 cagatgctgt caaaagaaag aaagaaaact catgtgaacc agaatggtgc atacaatcat
      601 ccattgctac cgagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca
      661 tttgcttctg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc atggtgagca
      721 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      781 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      841 ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      901 ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
      961 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1021 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1081 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1141 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1201 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
     1261 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     1321 accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1381 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa gtcgacggcg
     1441 cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat
     1501 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     1561 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     1621 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     1681 ctcggctgtt gggcactgac aattccgtgg ctcgagcgac tgtgccttct agttgccagc
     1741 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     1801 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     1861 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     1921 actagtccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     1981 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agagagggac
     2041 agatccgggc ccgcatgcgt cgacaattca ctggccgtcg ttttacaacg tcgtgactgg
     2101 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     2161 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     2221 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     2281 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     2341 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     2401 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     2461 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     2521 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2581 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2641 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     2701 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2761 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2821 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2881 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     2941 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3001 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3061 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3121 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3181 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     3241 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3301 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3361 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     3421 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     3481 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     3541 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3601 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3661 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3721 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3781 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3841 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3901 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3961 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4021 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4081 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4141 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4201 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4261 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4321 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4381 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     4441 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     4501 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     4561 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     4621 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     4681 tatgaccatg attacgccaa gctctcgaga tctag
//
