LOCUS       sequence-466233-        5017 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2065m 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 5017)
  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 5017)
  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: AiP1280 - pAAV-AiE2065m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5017
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   371..500
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(562..1150)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(642..661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1321..2112,2113..2181))
                     /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     1944..1963
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2182..2286)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2354..2372
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2410..2432)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2532..2551
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2568..3023)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2705..2726)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2917..2936
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3098..3238
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3109..3238
                     /label=AAV2 ITR
     polyA_signal    3356..3467
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3654..3674
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3767..4477,4478..4480,4481..4483))
                     /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(3809..3830)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4158..4177
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4417..4438
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4480..4489
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tatgtgggga gtggatgtgt gctgtctttc cttgggaagg acagcagaga gccaatcgtg
       61 gcctgtagcc taacaaatta tcttctcaag gagtagcctt aggccataag catgctcata
      121 tcacggagta ccagtaggag atggaacgac tactggtgct gaacataaac cccaagtgct
      181 tgatagaatt tttaaaacaa aatgaatcct atatatcttt caaggtgaaa ctaaatgttt
      241 ctatttgcat gggtcatgca aatctgacat agtcttacag gttgtaccag tgggttattc
      301 aataagaatg gtagcagcac aggcagaccg tgagctctct gcaagtttaa gaagcgacgc
      361 gtgcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
      421 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg
      481 agcgcgcagc tgcctgcagg acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      541 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      601 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
      661 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
      721 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
      781 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
      841 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
      901 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
      961 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1021 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1081 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1141 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1201 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1261 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1321 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1381 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     1441 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     1501 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     1561 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     1621 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     1681 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     1741 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     1801 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     1861 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     1921 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     1981 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     2041 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     2101 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     2161 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     2221 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     2281 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     2341 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     2401 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     2461 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     2521 gcttaactat gcggcatcag agcagattgt actgagagtg caccataaaa ttgtaaacgt
     2581 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2641 gaccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag agttgagtgt
     2701 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2761 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat caagtttttt
     2821 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2881 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2941 cgctaaggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3001 taatgcgccg ctacagggcg cgtactatgg ttgctttgac gtatgcggtg tgaaataccg
     3061 cacagatgcg taaggagaaa ataccgcatc aggcgcccct gcaggcagct gcgcgctcgc
     3121 tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc
     3181 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
     3241 ggccgctcgg tccgcacgtg agatctcatg cctgctattg tcttcccaat cctccccctt
     3301 gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac aatgcgatgc
     3361 aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg tcaaggaagg
     3421 cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg aagatctctc
     3481 gagccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg
     3541 cggcgatgag ttccgccgtg gcaagaacta accaggattt atacaaggag gagaaaatga
     3601 aagccatacg ggaagcaata gcatgataca aaggcattaa agcagcgtat ccacatagcg
     3661 taaaaggagc aacatagtta agaataccag tcaatctttc acaaattttg taatccagag
     3721 gttgattatg atatcgaatt cgcggccgcg gcgcgccgtc gacttacttg tacagctcgt
     3781 ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct
     3841 tctcgttggg gtctttgctc agcttggact ggtagctcag gtagtggttg tcgggcagca
     3901 gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgccgc
     3961 cgtcctcgat gttgtggcgg atcttgaagt tggccttgat gccgttcttc tgcttgtcgg
     4021 cggtgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc
     4081 cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga
     4141 acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct
     4201 ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt
     4261 agcgggcgaa gcactgcacg ccgtagccca gggtggtcac gagggtgggc cagggcacgg
     4321 gcagcttgcc ggtggtgcag atcagcttca gggtcagctt gccgtaggtg gcatcgccct
     4381 cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga
     4441 tgggcaccac cccggtgaac agctcctcgc ccttgctcac catggtggcc acgccagaag
     4501 caaatgtaag caatagatgg ctctgccctg acttttatgc ccagcccgag ctccttaaga
     4561 cataacatca tctggttctc taactttttc tttgggctga gtttggacca gaaacttgat
     4621 ttttagtatg tattttgttg tcaagtcaac acattcacaa ggcatttcaa acttttctta
     4681 cagtctctag aaattaggta ttttgttcag tttcactaga gcaacttcgc cttgcctcta
     4741 agtaggcccc tcagaagaat ggaaccagag tgccacttag tcagtaattg ctgctacgtg
     4801 ctgatggtgc cctggtggcc tccagaattg actgcagctt ccaaaagtgc cctgcatgca
     4861 accagaaagc attgtgactc cttggagaac tgagctcagt ggaaattact gaaaacaatt
     4921 gtcttcagct gagctgttat ataagcatgg actgaccagt cctgcaatcc agtcttgtac
     4981 aacagttcac ataatacttt gtgaaatata cggcctg
//
