LOCUS       sequence-466355-        4791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3054m 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 4791)
  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 4791)
  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: AiP20044 - pAAV-AiE3054m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   258..387
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(449..1037)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(529..548)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1208..1999,2000..2068))
                     /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     1831..1850
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2069..2173)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2241..2259
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2297..2319)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2419..2438
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2455..2910)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2592..2613)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2804..2823
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2985..3125
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2996..3125
                     /label=AAV2 ITR
     polyA_signal    3243..3354
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3541..3561
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3654..4364,4365..4367,4368..4370))
                     /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(3696..3717)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4045..4064
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4304..4325
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4367..4376
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 agccaggagg ctctgacaga gccagagcca catctgactt ccaggcactt cagaggaaaa
       61 tactagagtt cacccgggcc cctaagtctt tagcccaggg tcaaagcagg acatcttcag
      121 aggagttgaa aagggcaaca gtgtcaccaa agccaggggg aagagggcat gcttccagaa
      181 ggcagctaag cccatggaag gatctgtagt taaaaacata cccaggaagc acataagaga
      241 aagacgcgtg cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
      301 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga
      361 gcgagcgagc gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa aaggccagga
      421 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
      481 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
      541 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
      601 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
      661 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
      721 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
      781 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
      841 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
      901 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
      961 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1021 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1081 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1141 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1201 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1261 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     1321 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     1381 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     1441 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     1501 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     1561 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     1621 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     1681 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     1741 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     1801 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     1861 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     1921 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     1981 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     2041 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2101 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2161 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     2221 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     2281 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     2341 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     2401 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg
     2461 taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     2521 accaatagac cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagagt
     2581 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     2641 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa
     2701 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     2761 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     2821 gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     2881 ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga
     2941 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg
     3001 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
     3061 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     3121 ttcctgcggc cgctcggtcc gcacgtgaga tctcatgcct gctattgtct tcccaatcct
     3181 cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta ctcagacaat
     3241 gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca
     3301 aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc acagtcgaag
     3361 atctctcgag ccacggaatt gtcagtgccc aacagccgag cccctgtcca gcagcgggca
     3421 aggcaggcgg cgatgagttc cgccgtggca agaactaacc aggatttata caaggaggag
     3481 aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc agcgtatcca
     3541 catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca aattttgtaa
     3601 tccagaggtt gattatgata tcgaattcgc ggccgcggcg cgccgtcgac ttacttgtac
     3661 agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga
     3721 tcgcgcttct cgttggggtc tttgctcagc ttggactggt agctcaggta gtggttgtcg
     3781 ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc
     3841 acgccgccgt cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc
     3901 ttgtcggcgg tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg
     3961 atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg
     4021 ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg
     4081 cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg
     4141 tcggggtagc gggcgaagca ctgcacgccg tagcccaggg tggtcacgag ggtgggccag
     4201 ggcacgggca gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc gtaggtggca
     4261 tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg
     4321 accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc
     4381 ggtagcgcta gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat agatggctct
     4441 gccctgactt ttatgcccag cccgagctcc gccggcggtc aaatgagact aggttcatat
     4501 ctcagccctg tcatccagtg cagaatggtg taaggaaatt taatcttcaa gaactcccgg
     4561 attttctgta aaacaagatg aggagattgc tacctcacag gacataatga tcaaccaaaa
     4621 ccacacagaa aatagcgcgc acgagaaaat cacacagtgt tatctacatc tctaccgtca
     4681 ttgtgatcaa cgtcaccgtc agcagtcagt gcttcaaaca aacgaatgcg actgtgcaaa
     4741 gggttggatg cagaggatta aaaaatgagt aagattccat tcttgtcatt g
//
