LOCUS       sequence-470549-        4701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0698m 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 4701)
  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 4701)
  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: AiP12870 - pAAV-AiE0698m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      373..382
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(379..381,382..384,385..1095)
                     /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(424..445)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(685..704)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1032..1053
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1188..1208)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1395..1506
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1624..1764
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1624..1753
                     /label=AAV2 ITR
     rep_origin      1839..2294
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1926..1945)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2136..2157
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2311..2330)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2430..2452
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2490..2508)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2576..2680
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2681..2749,2750..3541)
                     /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(2899..2918)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3712..4300
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4201..4220
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4362..4491
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 atgtcaaggc agaatcttca cataaagcat gcattttgct ccctgttttc aatggtttta
       61 ctctgtagag tttctcaaat taattcaata cccactttaa attaaatgga aactttaatg
      121 ctatggtttg ctgtgccatt tagattttat gtgttgagat gcaattccaa ctctacttgg
      181 cacgaagtat agacactgaa tccaactatg atgtataagc aagggtcctt gtgaggtagg
      241 taatgagaat ggagcctgga acgtgtaaca ctggtgactg agctcgggct gggcataaaa
      301 gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct ttcgaagcta
      361 gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      421 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      481 gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg
      541 cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc
      601 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      661 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      721 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      781 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc
      841 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac
      901 ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
      961 ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag
     1021 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1081 gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc gaattcgata tcataatcaa
     1141 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     1201 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     1261 ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc
     1321 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct
     1381 cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1441 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1501 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1561 caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc
     1621 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     1681 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     1741 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     1801 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     1861 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     1921 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     1981 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2041 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2101 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2161 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     2221 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     2281 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2341 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     2401 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     2461 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     2521 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     2581 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2641 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2701 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2761 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2821 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     2881 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     2941 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3001 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3061 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3121 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3181 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3241 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3301 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3361 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3421 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3481 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3541 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3601 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3661 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3721 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3781 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3841 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3901 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3961 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4021 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4081 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4141 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4201 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4261 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4321 tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc
     4381 tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     4441 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca
     4501 cgcgttgtat gcctaaaggg tttgcttcta tgttaattcc caacaacaat catgatactt
     4561 ccttacttct gcaggactag gctggctcac tagtgaggtt ttctccaagg atgtgctcca
     4621 atatctgctc cacaatactt aaatagcttc acaatacata aatatttaag tactcctctg
     4681 taaggagcgt tctgtttcca t
//
