LOCUS       sequence-461898-        5159 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0082m 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 5159)
  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 5159)
  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: AiP11576 -
            pAAV-hsA2-AiE0082m-minRho-SYFP2-WPR...
FEATURES             Location/Qualifiers
     source          1..5159
                     /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      1196..1205
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1202..1204,1205..1207,1208..1918)
                     /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(1247..1268)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1508..1527)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1855..1876
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1986..2006)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2193..2304
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2422..2562
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2422..2551
                     /label=AAV2 ITR
     rep_origin      2637..3092
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2724..2743)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2934..2955
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3109..3128)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3228..3250
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3288..3306)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3374..3478
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3479..3547,3548..4339)
                     /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(3697..3716)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4510..5098
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4999..5018
                     /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 gcggccgcaa cgcgtttaga acaatggctg gcccatagta aatgccgtgt
      181 tagtgtgtta gttgctgttc ttccacgtca gaagaggcac agacaaatta ccaccaggtg
      241 gcgctcagag tctgcggagg catcacaaca gccctgaatt tgaatcctgc tctgccactg
      301 cctagttgag accttttact acctgactag ctgtttgtgt attttaggtg tttgttttca
      361 gcacacacta tggatcacag ctcaataacc cctaggtatc aataccagac atctctcagt
      421 ctgacgaccc catcccccaa gccccaattg ttctattctg tccaattact tatcatgagg
      481 acatggaaga agggcaaacc tgggcagcaa tgagagccaa agtctacttc agccagaatc
      541 aaagctattg gagaatgaaa ggaaagaaga gggtggtatg cgtgtaaaag gacgggatgg
      601 cgtctggctt gggactggat ctcagctgta taccgcagtg ccctctgcct ctgcagcatg
      661 cgtccctgtg gtgggactgg gggtaggact ggatctgctt agcaggcacc tggggaagga
      721 ctccaattcc gtttctctct ggactgcagc attctgctca gctgcaggct ggatgaacta
      781 aaatgacctt gggtaagaaa acagccacgg caattcataa tacagctgac tttggccttg
      841 tgccgagcag tcgaggtgaa ccagccacag ttcctgacca cgcctgcagt tacctccgct
      901 tacttgaact tgaataaaca caccttccag ctgcacccct ttctgaacaa gtcctgaaag
      961 ttaaggctgt tcctgaagcc actctcaggc taaagaggca tgcgaattcg atatcataat
     1021 caaccatagg taccgagctc gggattcagc cgggagctta gggaggggag gtcacttcat
     1081 aagggcctgg ggggggagtt ggagccacga gtcgtccagc cggagccccg tgtggctgag
     1141 ctccggcctc agaagcatcc ccgggttgga tccttcgaag ctagcgctac cggtcgccac
     1201 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1261 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1321 cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac
     1381 cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa
     1441 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     1501 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1561 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1621 caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa
     1681 cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc
     1741 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1801 ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     1861 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     1921 agtcgacatc ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     1981 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2041 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc
     2101 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2161 actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt
     2221 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2281 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2341 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc
     2401 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     2461 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     2521 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     2581 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     2641 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2701 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2761 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2821 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     2881 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2941 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     3001 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgg
     3061 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     3121 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     3181 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3241 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     3301 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     3361 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3421 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3481 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3541 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3601 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3661 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3721 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3781 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3841 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3901 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3961 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4021 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4081 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4141 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4201 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4261 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4321 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4381 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4441 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4501 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4561 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4621 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4681 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4741 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4801 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4861 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4921 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4981 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5041 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5101 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
