LOCUS       sequence-466324-        4873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2561m 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 4873)
  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 4873)
  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: AiP1902 - pAAV-AiE2561m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      331..340
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(337..339,340..342,343..1053)
                     /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(382..403)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(643..662)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     990..1011
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1146..1166)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1353..1464
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1582..1722
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1582..1711
                     /label=AAV2 ITR
     rep_origin      1797..2252
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1884..1903)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2094..2115
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2269..2288)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2388..2410
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2448..2466)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2534..2638
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2639..2707,2708..3499)
                     /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(2857..2876)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3670..4258
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4159..4178
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4320..4449
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cagggtgtgt ttgttcatgg cagtgtgttc tctccatctg tctgcagaag agatctacgg
       61 taactggagt atgcctctca acacctccca tatctggcat cctcgtatcc gggaggtggg
      121 tgaagagatt gggtttgaag gaagggctgg gtgggcaggc cactgctaag ctgtatgcaa
      181 ggaggtggct gaggaactta catattgata atgccaggtc aaagtgacta tagctcggag
      241 ctcgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg cttctgggat
      301 ccagatcttt cgaagctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg
      361 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      421 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc
      481 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc
      541 gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
      601 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
      661 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
      721 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
      781 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc
      841 cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
      901 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg
      961 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1021 gggatcactc tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga
     1081 attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     1141 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     1201 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc
     1261 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     1321 actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt
     1381 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     1441 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     1501 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc
     1561 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     1621 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     1681 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     1741 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     1801 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1861 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1921 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1981 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     2041 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2101 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     2161 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2221 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     2281 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     2341 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     2401 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     2461 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     2521 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2581 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     2641 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     2701 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     2761 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     2821 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     2881 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     2941 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3001 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3061 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3121 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     3181 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3241 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3301 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     3361 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     3421 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     3481 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     3541 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3601 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3661 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3721 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3781 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3841 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3901 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3961 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4021 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4081 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4141 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4201 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4261 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtc
     4321 ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg
     4381 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
     4441 ggggttcctg cggccgcaac aagtctgttg ggttcagtgg tggtagctgg gaacaatcgc
     4501 aggcagccct cagattcaga aatggagcag tctgctttgg agcacatgtc tcttcttggc
     4561 actgaggtgg gggagctgaa ggcaagatgc agggtttgaa ctggaggatc cctgaaaatg
     4621 tgggggcggg actccatgtg gggtcttagg gacttccctt tatccctttt cctcaaccca
     4681 ctccaccact gctatgtgcc ccgccttcca ggctgccgtt gttccaggcc agtgcctttg
     4741 cctttctggt tccagccaag tcaatcctgg ccttggagag gtggaagtgt ccttcagaag
     4801 gtgggttcag tttgcccatc tgcactggtg gagaggtcgt gaaggtctgg gcagcttccc
     4861 agagtctagg cac
//
