LOCUS       sequence-433045-        4884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2386m 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 4884)
  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 4884)
  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: AiP1731 - pAAV-AiE2386m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(438..1229,1230..1298))
                     /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     1061..1080
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1299..1403)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1471..1489
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1527..1549)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1649..1668
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1685..2140)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1822..1843)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2034..2053
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2215..2355
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2226..2355
                     /label=AAV2 ITR
     polyA_signal    2473..2584
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2771..2791
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2884..3594,3595..3597,3598..3600))
                     /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(2926..2947)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3275..3294
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3534..3555
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3597..3606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4372..4501
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4563..4884,1..267))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4643..4662)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
       61 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
      121 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
      181 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
      241 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
      301 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
      361 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
      421 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
      481 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
      541 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
      601 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
      661 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
      721 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
      781 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
      841 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
      901 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
      961 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     1021 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     1081 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     1141 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     1201 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     1261 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     1321 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     1381 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     1441 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     1501 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     1561 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     1621 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     1681 cataaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     1741 tcatttttta accaatagac cgaaatcggc aaaatccctt ataaatcaaa agaatagccc
     1801 gagatagagt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     1861 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     1921 cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     1981 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     2041 aaagcgaaag gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     2101 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgta
     2161 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca
     2221 ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga
     2281 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
     2341 tcactagggg ttcctgcggc cgctcggtcc gcacgtgaga tctcatgcct gctattgtct
     2401 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
     2461 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
     2521 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
     2581 acagtcgaag atctctcgag ccacggaatt gtcagtgccc aacagccgag cccctgtcca
     2641 gcagcgggca aggcaggcgg cgatgagttc cgccgtggca agaactaacc aggatttata
     2701 caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc
     2761 agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca
     2821 aattttgtaa tccagaggtt gattatgata tcgaattcgc ggccgcggcg cgccgtcgac
     2881 ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag
     2941 gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggt agctcaggta
     3001 gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc
     3061 ggcgagctgc acgccgccgt cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc
     3121 gttcttctgc ttgtcggcgg tgatatagac gttgtggctg ttgtagttgt actccagctt
     3181 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     3241 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     3301 gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     3361 cttcatgtgg tcggggtagc gggcgaagca ctgcacgccg tagcccaggg tggtcacgag
     3421 ggtgggccag ggcacgggca gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc
     3481 gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     3541 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
     3601 ggtggcgacc ggtagcgcta gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat
     3661 agatggctct gccctgactt ttatgcccag cccgagctcc gactctagct gagaagaaga
     3721 tgaattaagg agactgttga agtcatgtga atttgaggtc acaaagacca gtcgttagtt
     3781 aacctagtac cgagaaactg aagaaagatg tctatggatt tcatttatat tattagtgcc
     3841 gacttagtga tttgattttt ctctgctatg gatatataag gaactcctgt ctcatgcttt
     3901 ctattccaaa caactataat gatgtacact caacttttta agtttaaatg aaacattttt
     3961 ttaaaaaaat gaaacacttt tgtccctgct tagctttttt cagacttctg aaactcttaa
     4021 atgttcttat tttcatagca ctgtagttac atgcatacat tcacattatg taatatagtt
     4081 atttgcatgt ccttgttaaa ataacataat tagaaacatt ggttatacaa ccaaggtctt
     4141 actcgtaatg tcatatttaa ggatgatgtt tctttaatcc accgcaatca cacattttag
     4201 gaaagtaagt tttaatgtta tagaatcaat acctgcaaat ccttcttaga aaattaactc
     4261 aaacctaatt acaagatccc agcctaagag gtgggcaagg tcagtaactt gtaggccagg
     4321 cactcctagg acatttgggg cgattattac tcttacaacg cgtgcggccg caggaacccc
     4381 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     4441 caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     4501 gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4561 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4621 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4681 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4741 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4801 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4861 aactatcgtc ttgagtccaa cccg
//
