LOCUS       sequence-466250-        4913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2149m 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 4913)
  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 4913)
  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: AiP1378 - pAAV-AiE2149m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(39..627)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(119..138)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(798..1589,1590..1658))
                     /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     1421..1440
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1659..1763)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1831..1849
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1887..1909)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2009..2028
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2045..2500)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2182..2203)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2394..2413
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2575..2715
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2586..2715
                     /label=AAV2 ITR
     polyA_signal    2833..2944
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3131..3151
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3244..3954,3955..3957,3958..3960))
                     /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(3286..3307)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3635..3654
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3894..3915
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3957..3966
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4761..4890
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
       61 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      121 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
      181 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
      241 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
      301 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      361 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
      421 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
      481 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
      541 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
      601 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
      661 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
      721 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
      781 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
      841 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
      901 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
      961 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     1021 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     1081 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     1141 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     1201 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     1261 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     1321 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     1381 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     1441 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     1501 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     1561 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     1621 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     1681 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     1741 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     1801 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     1861 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     1921 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     1981 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     2041 cataaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     2101 tcatttttta accaatagac cgaaatcggc aaaatccctt ataaatcaaa agaatagccc
     2161 gagatagagt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     2221 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     2281 cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     2341 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     2401 aaagcgaaag gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     2461 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgta
     2521 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca
     2581 ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga
     2641 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
     2701 tcactagggg ttcctgcggc cgctcggtcc gcacgtgaga tctcatgcct gctattgtct
     2761 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
     2821 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
     2881 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
     2941 acagtcgaag atctctcgag ccacggaatt gtcagtgccc aacagccgag cccctgtcca
     3001 gcagcgggca aggcaggcgg cgatgagttc cgccgtggca agaactaacc aggatttata
     3061 caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc
     3121 agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca
     3181 aattttgtaa tccagaggtt gattatgata tcgaattcgc ggccgcggcg cgccgtcgac
     3241 ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag
     3301 gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggt agctcaggta
     3361 gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc
     3421 ggcgagctgc acgccgccgt cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc
     3481 gttcttctgc ttgtcggcgg tgatatagac gttgtggctg ttgtagttgt actccagctt
     3541 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     3601 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     3661 gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     3721 cttcatgtgg tcggggtagc gggcgaagca ctgcacgccg tagcccaggg tggtcacgag
     3781 ggtgggccag ggcacgggca gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc
     3841 gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     3901 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
     3961 ggtggcgacc ggtagcgcta gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat
     4021 agatggctct gccctgactt ttatgcccag cccgagctcg ggagaataca cttccatttc
     4081 tcagatgttc tgggaacttg ccttgctgtg ctgtgatcat aggctgtgtt ttcctaggac
     4141 aattgtcttg catttgtggg aaaggatacg aagatccttc cttttctccc tgaaggtcca
     4201 attttctttt ttaaacagga tctgcccttc cactcttccc acagttatta tcctgatgtc
     4261 aacagcagag aaagttaaaa ggtcttttct cactgggacc ttgaagcact aatttgcagc
     4321 atttatggta aaaagccctg ctgcaagcct ttttctggcg catgtatttc cacactctga
     4381 cctttcagaa ctggccttta agaaggaaca gttgtcagga atgggaagta atacggacaa
     4441 tccacaaatt tttaccattt tctgtcaaaa ttgacctctt gatcatctct cagaggcctt
     4501 ctgatgatga ccagccacct aagggaatta cagctgccag gatttcttgt agaaaatgtc
     4561 tagaaaggtc tgtactcagt ctcacctctc ctactacatc tgttactcat aataaacaac
     4621 tccttaaagt gctcctgcag ctaggactgc ccttcttttc tgaaaatgac cattggccag
     4681 atatagaagg aacccaaatt tagagaggga aatgaagtta cttaggcaag tttgggagcc
     4741 ttctaaacgc gtgcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     4801 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag
     4861 tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc aaaaggccag caa
//
