LOCUS       sequence-470540-        4499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0726m 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 4499)
  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 4499)
  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: AiP12865 - pAAV-AiE0726m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(298..1089,1090..1158))
                     /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     921..940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1159..1263)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1331..1349
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1387..1409)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1509..1528
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1545..2000)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1682..1703)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1894..1913
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2075..2215
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2086..2215
                     /label=AAV2 ITR
     polyA_signal    2333..2444
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2631..2651
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2744..3454,3455..3457,3458..3460))
                     /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(2786..2807)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3135..3154
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3394..3415
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3457..3466
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3847..3976
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4038..4499,1..127))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4118..4137)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
       61 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      121 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
      181 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
      241 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
      301 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
      361 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
      421 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
      481 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
      541 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
      601 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
      661 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
      721 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
      781 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
      841 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
      901 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
      961 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1021 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1081 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1141 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     1201 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     1261 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     1321 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     1381 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     1441 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     1501 taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg taaacgttaa
     1561 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaatagac
     1621 cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagagt tgagtgttgt
     1681 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     1741 aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg
     1801 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     1861 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     1921 taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     1981 tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac
     2041 agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg cgctcgctcg
     2101 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
     2161 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc
     2221 cgctcggtcc gcacgtgaga tctcatgcct gctattgtct tcccaatcct cccccttgct
     2281 gtcctgcccc accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat
     2341 ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac
     2401 gggggagggg caaacaacag atggctggca actagaaggc acagtcgaag atctctcgag
     2461 ccacggaatt gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg
     2521 cgatgagttc cgccgtggca agaactaacc aggatttata caaggaggag aaaatgaaag
     2581 ccatacggga agcaatagca tgatacaaag gcattaaagc agcgtatcca catagcgtaa
     2641 aaggagcaac atagttaaga ataccagtca atctttcaca aattttgtaa tccagaggtt
     2701 gattatgata tcgaattcgc ggccgcggcg cgccgtcgac ttacttgtac agctcgtcca
     2761 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     2821 cgttggggtc tttgctcagc ttggactggt agctcaggta gtggttgtcg ggcagcagca
     2881 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt
     2941 cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg
     3001 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     3061 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     3121 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     3181 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     3241 gggcgaagca ctgcacgccg tagcccaggg tggtcacgag ggtgggccag ggcacgggca
     3301 gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     3361 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     3421 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtagcgcta
     3481 gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat agatggctct gccctgactt
     3541 ttatgcccag cccgagctct gaagaagctg ggtttggcat tacatgtgat ggaaaagtaa
     3601 gaatgggtac atttaaaata tcattctact ttctatgaga agaaacaaac aggagccagg
     3661 aagaaccaaa ggaaacagcc agtgccagag tgggaaggca gagctggaga cagttggatg
     3721 gttatgggta tgatttgata tgggaggatt cttggatcta tttctagtgt tgggcacagg
     3781 taggatgagt tagagaagct tgccctggta tgggcaacaa gaaacacaca gaacgcgtgc
     3841 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3901 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg
     3961 cgcagctgcc tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4021 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4081 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4141 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4201 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4261 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4321 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4381 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4441 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgc
//
