LOCUS       sequence-432574-        4735 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2566m 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 4735)
  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 4735)
  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: AiP1862 - pAAV-AiE2566m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4735
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     124..145
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(299..318)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     418..440
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(478..496)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        564..668
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(669..737,738..1529)
                     /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(887..906)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1700..2288
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2189..2208
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2350..2479
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3096..3105
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3102..3104,3105..3107,3108..3818)
                     /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(3147..3168)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3408..3427)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3755..3776
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3911..3931)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4118..4229
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4347..4487
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4347..4476
                     /label=AAV2 ITR
     rep_origin      join(4562..4735,1..282)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4649..4668)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
       61 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      121 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
      181 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
      241 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
      301 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
      361 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
      421 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
      481 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
      541 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
      601 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      661 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      721 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      781 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      841 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      901 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      961 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     1021 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     1081 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     1141 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     1201 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     1261 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     1321 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     1381 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     1441 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     1501 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1561 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1621 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1681 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1741 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1801 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1861 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1921 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1981 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2041 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2101 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2161 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     2221 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     2281 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     2341 ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg
     2401 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
     2461 tccatcacta ggggttcctg cggccgcacg cgttgtaaga gtaataatcg gaaacggctt
     2521 cctgagtgtt tccaaggaga gacccttgag cccccatggt gttcttcatg cccaccttgg
     2581 actttatggt atgttcttca catagaaggt atgatgcaag cctgtctagt gctgaagcac
     2641 cctgcccatg ggttcttccc agtgccagac tcttgcagtc tgcctgcagc cacatggctt
     2701 tctaactcta gcttcttggc tcttggtctc tctatgaact tgcagcatgt tgaatgccaa
     2761 ggcagcagac ttcactgaca gatttttgct caccacgtgg ccaggaagga tctgggcatg
     2821 tccttcaatg ggatatagac atcctgtcct cctgctggca gaaatcagtg gcacctaagt
     2881 ttcttcctct cctgaagtgg caggcctggc tctccacccg tggagcagga acagctgtgg
     2941 atctgtggga aaggaagagt gctgattttc tgtcccagtt ttgcttggaa actgagtggt
     3001 cggagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct
     3061 gggatccaga tctttcgaag ctagcgctac cggtcgccac catggtgagc aagggcgagg
     3121 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     3181 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc
     3241 tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct
     3301 acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     3361 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     3421 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     3481 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     3541 acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     3601 agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca
     3661 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca
     3721 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     3781 ccgccgggat cactctcggc atggacgagc tgtacaagta agtcgacggc gcgccgcggc
     3841 cgcgaattcg atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     3901 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     3961 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc
     4021 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     4081 tgggcactga caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca
     4141 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     4201 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     4261 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag
     4321 atctcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
     4381 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     4441 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     4501 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     4561 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4621 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4681 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttcc
//
