LOCUS       sequence-466313-        5009 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2429m 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 5009)
  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 5009)
  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: AiP1855 - pAAV-AiE2429m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(31..50)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(710..1501,1502..1570))
                     /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     1333..1352
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1571..1675)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1743..1761
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1799..1821)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1921..1940
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1957..2412)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2094..2115)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2306..2325
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2487..2627
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2498..2627
                     /label=AAV2 ITR
     polyA_signal    2745..2856
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3043..3063
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3156..3866,3867..3869,3870..3872))
                     /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(3198..3219)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3547..3566
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3806..3827
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3869..3878
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4769..4898
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4960..5009,1..539))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
       61 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
      121 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
      181 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
      241 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
      301 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
      361 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
      421 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
      481 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
      541 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
      601 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
      661 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
      721 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
      781 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
      841 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
      901 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
      961 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     1021 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     1081 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     1141 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     1201 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     1261 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     1321 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     1381 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     1441 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     1501 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     1561 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     1621 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     1681 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     1741 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     1801 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     1861 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     1921 cggcatcaga gcagattgta ctgagagtgc accataaaat tgtaaacgtt aatattttgt
     1981 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag accgaaatcg
     2041 gcaaaatccc ttataaatca aaagaatagc ccgagataga gttgagtgtt gttccagttt
     2101 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     2161 atcagggcga tggcccacta cgtgaaccat cacccaaatc aagttttttg gggtcgaggt
     2221 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     2281 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctaaggcgc
     2341 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     2401 tacagggcgc gtactatggt tgctttgacg tatgcggtgt gaaataccgc acagatgcgt
     2461 aaggagaaaa taccgcatca ggcgcccctg caggcagctg cgcgctcgct cgctcactga
     2521 ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga
     2581 gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg gccgctcggt
     2641 ccgcacgtga gatctcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc
     2701 ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat
     2761 tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg
     2821 ggcaaacaac agatggctgg caactagaag gcacagtcga agatctctcg agccacggaa
     2881 ttgtcagtgc ccaacagccg agcccctgtc cagcagcggg caaggcaggc ggcgatgagt
     2941 tccgccgtgg caagaactaa ccaggattta tacaaggagg agaaaatgaa agccatacgg
     3001 gaagcaatag catgatacaa aggcattaaa gcagcgtatc cacatagcgt aaaaggagca
     3061 acatagttaa gaataccagt caatctttca caaattttgt aatccagagg ttgattatga
     3121 tatcgaattc gcggccgcgg cgcgccgtcg acttacttgt acagctcgtc catgccgaga
     3181 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg
     3241 tctttgctca gcttggactg gtagctcagg tagtggttgt cgggcagcag cacggggccg
     3301 tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgccgcc gtcctcgatg
     3361 ttgtggcgga tcttgaagtt ggccttgatg ccgttcttct gcttgtcggc ggtgatatag
     3421 acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg
     3481 aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg
     3541 gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct
     3601 tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcgggcgaag
     3661 cactgcacgc cgtagcccag ggtggtcacg agggtgggcc agggcacggg cagcttgccg
     3721 gtggtgcaga tcagcttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg
     3781 gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc
     3841 ccggtgaaca gctcctcgcc cttgctcacc atggtggcga ccggtagcgc tagcttcgaa
     3901 agatctggat cccagaagca aatgtaagca atagatggct ctgccctgac ttttatgccc
     3961 agcccgagct ccgctcattt attttcctgt aaatgcttct attgctaacc aggccttggc
     4021 catttttccc atgggaaatg gagttgttgg cttcccctgg ttcctgttca ccatccagga
     4081 agtcgaataa ataatgaccg tagcccatac tggtgggatg gatacaggag gaggactgaa
     4141 agcaggagaa tatcaaaccc atcaaaaatc accgagatca gtgcaaagac tgaagcagaa
     4201 agacacagcc tctaaatagg agcggcctta atgaaagtgc atcacctcag agccaacatg
     4261 aagtctgcag atgggatgca cctgcctcct gagaacctca cagagggaaa tcaagcatca
     4321 gaacccagaa tggaagcaga aaacccccaa tcagactcct cggagacact tcagaaaagg
     4381 aactctcagc ttgcaaaaca acaggtgcat ctctaaagag atgtggctcc tgcccccacc
     4441 ccataccacc gcaccctgag attgcagcac tgctctgaac tctccaatca agcctctttc
     4501 tttggtcact cttaaagtgg ttggtttcca gcactaaccc tcactcctcc accgaaatga
     4561 aatctgctag ctttggctga tgtttaaaca gcttcaatct ctggggtctt ttgtgagaga
     4621 ggataatttt aacactggtg ataaacccac cagaatttcc aggtgagagg agaagcctag
     4681 tggagccttt atccagctgt tgaagacagg acctgggaac tcttgggaca aagacatcga
     4741 ttattactct tacaacgcgt gcggccgcag gaacccctag tgatggagtt ggccactccc
     4801 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     4861 ggcctcagtg agcgagcgag cgcgcagctg cctgcaggac atgtgagcaa aaggccagca
     4921 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4981 tgacgagcat cacaaaaatc gacgctcaa
//
