LOCUS       sequence-438962-        4932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2347m 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 4932)
  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 4932)
  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: AiP1668 - pAAV-AiE2347m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   526..655
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(717..1305)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(797..816)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1476..2267,2268..2336))
                     /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     2099..2118
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2337..2441)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2509..2527
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2565..2587)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2687..2706
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2723..3178)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2860..2881)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3072..3091
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3253..3393
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3264..3393
                     /label=AAV2 ITR
     polyA_signal    3511..3622
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3809..3829
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3922..4632,4633..4635,4636..4638))
                     /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(3964..3985)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4313..4332
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4572..4593
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4635..4644
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gcactaaata ccactttaaa aaaaaaaaaa gcctcagtga gattttgatt agggaggcat
       61 cgggtttgtt cattaatttg gggcgcgttg acaacttcac aatatcgaat cttcaggacc
      121 ctaacatttc aagggtggtt gagtcagaaa ctctgattaa tgagagtggg gaggaaaagt
      181 cgtcatagag atggtttaaa aacaaggaga ggggggttgc catagaaacc aagagagcaa
      241 gagtttctga gaaggggagt gatcaacaat atcaaaccct gcagagaatt ccaataagac
      301 agggatggaa aaacacccac tggaggtggt aattaggggg ttatcggagc ccgcagcaag
      361 agtggttttt ggggtgtgat ggagcagaag cctgccagcg gcgctgcctg gggatgacag
      421 atagtcagca gcaaggcagc cctgtgcaga agtttagata attaagttca gataattaga
      481 aaagtttagg gcagcgatta ttactcttac aacgcgtgcg gccgcaggaa cccctagtga
      541 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
      601 tcgcccgacg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggacatg
      661 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      721 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      781 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      841 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      901 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      961 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1021 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1081 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1141 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1201 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1261 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1321 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1381 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1441 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1501 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1561 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1621 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1681 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1741 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1801 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1861 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1921 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1981 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     2041 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     2101 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     2161 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     2221 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     2281 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     2341 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     2401 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     2461 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     2521 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     2581 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     2641 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     2701 attgtactga gagtgcacca taaaattgta aacgttaata ttttgttaaa attcgcgtta
     2761 aatttttgtt aaatcagctc attttttaac caatagaccg aaatcggcaa aatcccttat
     2821 aaatcaaaag aatagcccga gatagagttg agtgttgttc cagtttggaa caagagtcca
     2881 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     2941 ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta
     3001 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3061 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg
     3121 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac
     3181 tatggttgct ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     3241 gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
     3301 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     3361 agggagtggc caactccatc actaggggtt cctgcggccg ctcggtccgc acgtgagatc
     3421 tcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
     3481 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
     3541 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
     3601 ggctggcaac tagaaggcac agtcgaagat ctctcgagcc acggaattgt cagtgcccaa
     3661 cagccgagcc cctgtccagc agcgggcaag gcaggcggcg atgagttccg ccgtggcaag
     3721 aactaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag caatagcatg
     3781 atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat agttaagaat
     3841 accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatgatatc gaattcgcgg
     3901 ccgcggcgcg ccgtcgactt acttgtacag ctcgtccatg ccgagagtga tcccggcggc
     3961 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt
     4021 ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     4081 gttctgctgg tagtggtcgg cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt
     4141 gaagttggcc ttgatgccgt tcttctgctt gtcggcggtg atatagacgt tgtggctgtt
     4201 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     4261 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     4321 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     4381 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact gcacgccgta
     4441 gcccagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatcag
     4501 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     4561 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     4621 ctcgcccttg ctcaccatgg tggcgaccgg tagcgctagc ttcgaaagat ctggatccca
     4681 gaagcaaatg taagcaatag atggctctgc cctgactttt atgcccagcc cgagctccgg
     4741 gatgccaaag tcttcaggat tcctctctgc accatttcca atctttaatc acagtcacgt
     4801 tgtcatctgt cgcctagtag ggcgaatcca gaccttttcc tcctactgtg ttcccaacct
     4861 tgtttttctt tctcaacatc atcttggctg tttttgccca tttccctcta ccatatggct
     4921 tttagaatca at
//
