LOCUS       sequence-466289-        4913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2360m 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: AiP1678 - pAAV-AiE2360m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   191..320
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(382..970)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(462..481)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1141..1932,1933..2001))
                     /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     1764..1783
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2002..2106)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2174..2192
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2230..2252)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2352..2371
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2388..2843)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2525..2546)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2737..2756
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2918..3058
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2929..3058
                     /label=AAV2 ITR
     polyA_signal    3176..3287
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3474..3494
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3587..4297,4298..4300,4301..4303))
                     /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(3629..3650)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3978..3997
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4237..4258
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4300..4309
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 aaataaccct gttaacaatg tggggggtgt gggggaggag agcagagctt gagtgccagc
       61 ccttcctgcc ttggaagtgc tgcgggcctg aggtgggggg acttggggct cctaggcaga
      121 ctcctgccct ctggaggagg ctgtgagcct tgcctgctcc gattattact cttacaacgc
      181 gtgcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
      241 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg
      301 agcgcgcagc tgcctgcagg acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      361 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      421 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
      481 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
      541 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
      601 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
      661 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
      721 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
      781 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
      841 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
      901 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
      961 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1021 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1081 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1141 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1201 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     1261 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     1321 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     1381 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     1441 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     1501 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     1561 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     1621 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     1681 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     1741 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     1801 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     1861 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     1921 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     1981 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     2041 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     2101 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     2161 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     2221 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     2281 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     2341 gcttaactat gcggcatcag agcagattgt actgagagtg caccataaaa ttgtaaacgt
     2401 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2461 gaccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag agttgagtgt
     2521 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2581 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat caagtttttt
     2641 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2701 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2761 cgctaaggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     2821 taatgcgccg ctacagggcg cgtactatgg ttgctttgac gtatgcggtg tgaaataccg
     2881 cacagatgcg taaggagaaa ataccgcatc aggcgcccct gcaggcagct gcgcgctcgc
     2941 tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc
     3001 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
     3061 ggccgctcgg tccgcacgtg agatctcatg cctgctattg tcttcccaat cctccccctt
     3121 gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac aatgcgatgc
     3181 aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg tcaaggaagg
     3241 cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg aagatctctc
     3301 gagccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg
     3361 cggcgatgag ttccgccgtg gcaagaacta accaggattt atacaaggag gagaaaatga
     3421 aagccatacg ggaagcaata gcatgataca aaggcattaa agcagcgtat ccacatagcg
     3481 taaaaggagc aacatagtta agaataccag tcaatctttc acaaattttg taatccagag
     3541 gttgattatg atatcgaatt cgcggccgcg gcgcgccgtc gacttacttg tacagctcgt
     3601 ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct
     3661 tctcgttggg gtctttgctc agcttggact ggtagctcag gtagtggttg tcgggcagca
     3721 gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgccgc
     3781 cgtcctcgat gttgtggcgg atcttgaagt tggccttgat gccgttcttc tgcttgtcgg
     3841 cggtgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc
     3901 cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga
     3961 acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct
     4021 ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt
     4081 agcgggcgaa gcactgcacg ccgtagccca gggtggtcac gagggtgggc cagggcacgg
     4141 gcagcttgcc ggtggtgcag atcagcttca gggtcagctt gccgtaggtg gcatcgccct
     4201 cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga
     4261 tgggcaccac cccggtgaac agctcctcgc ccttgctcac catggtggcg accggtagcg
     4321 ctagcttcga aagatctgga tcccagaagc aaatgtaagc aatagatggc tctgccctga
     4381 cttttatgcc cagcccgagc tccgctgtac tctgtctgta ttttctcccc tactgtgcta
     4441 cacccacatt tgcatatttt taatgagttt ctctattcct ccccattatt ctactgtggt
     4501 atatattgtt ggcagaatat tgaacccttg ggagtggccc agagctgctg aggaaagagg
     4561 tgacggtatt ttatcttatt agaccacagc cgggctcctg gccccaaact tggtaacagg
     4621 ctggcccgag ggaaagcaca gagggtgatt tcggctgctc gattctcagg catcttccaa
     4681 agatagagta aatggagttg gcgctgctgt tcaataacaa agcctctggc tggaaaggaa
     4741 gtagagacgc taaaaaggac cagatcacgt gatgatttgc agagggcctg cctgtcttcc
     4801 tagaaatttc ccaagactca acagaaaaac tgaaaacatt aaatagttga ctaggatggc
     4861 catgcacaaa cctgcttctc catttacagg tttactttga aaggaaaaaa aga
//
