LOCUS       sequence-466346-        4677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2665m 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 4677)
  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 4677)
  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: AiP20011 - pAAV-AiE2665m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(207..998,999..1067))
                     /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     830..849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1068..1172)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1240..1258
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1296..1318)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1418..1437
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1454..1909)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1591..1612)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1803..1822
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1984..2124
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1995..2124
                     /label=AAV2 ITR
     polyA_signal    2242..2353
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2540..2560
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2653..3363,3364..3366,3367..3369))
                     /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(2695..2716)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3044..3063
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3303..3324
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3366..3375
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3934..4063
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4125..4677,1..36))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4205..4224)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
       61 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
      121 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
      181 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
      241 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
      301 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
      361 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
      421 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
      481 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
      541 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
      601 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
      661 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
      721 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
      781 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
      841 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
      901 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
      961 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     1021 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     1081 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     1141 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     1201 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     1261 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     1321 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     1381 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     1441 agagtgcacc ataaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     1501 taaatcagct cattttttaa ccaatagacc gaaatcggca aaatccctta taaatcaaaa
     1561 gaatagcccg agatagagtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     1621 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     1681 gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     1741 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     1801 gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg
     1861 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta ctatggttgc
     1921 tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     1981 gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc
     2041 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
     2101 ccaactccat cactaggggt tcctgcggcc gctcggtccg cacgtgagat ctcatgcctg
     2161 ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc cagaatagaa
     2221 tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag gacagtggga
     2281 gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga tggctggcaa
     2341 ctagaaggca cagtcgaaga tctctcgagc cacggaattg tcagtgccca acagccgagc
     2401 ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa gaactaacca
     2461 ggatttatac aaggaggaga aaatgaaagc catacgggaa gcaatagcat gatacaaagg
     2521 cattaaagca gcgtatccac atagcgtaaa aggagcaaca tagttaagaa taccagtcaa
     2581 tctttcacaa attttgtaat ccagaggttg attatgatat cgaattcgcg gccgcggcgc
     2641 gccgtcgact tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
     2701 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct tggactggta
     2761 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     2821 gtagtggtcg gcgagctgca cgccgccgtc ctcgatgttg tggcggatct tgaagttggc
     2881 cttgatgccg ttcttctgct tgtcggcggt gatatagacg ttgtggctgt tgtagttgta
     2941 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     3001 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     3061 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     3121 gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac tgcacgccgt agcccagggt
     3181 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatca gcttcagggt
     3241 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     3301 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     3361 gctcaccatg gtggcgaccg gtagcgctag cttcgaaaga tctggatccc agaagcaaat
     3421 gtaagcaata gatggctctg ccctgacttt tatgcccagc ccgagctccg ccggcgggca
     3481 gaaatctggg ctcccgagac cctacaagac agccagcaga cctgtggtgc taggtgagct
     3541 gtgggactct ggcgcctgtc actttccctg tctctggatt gcgctggcaa ttccaaggct
     3601 ctcggagact cctttccgtg tttacatcat ttagcaggga aggactgtta atgactaatc
     3661 tgtgtccatg gggcacagag ccaagggaga gatgctgaag cttatgccgg aaggccgcct
     3721 gtggtcaccc aggcttctgc ttcagtctct cctccttctc ctcctcagcc cactgtctcc
     3781 tcatctcacc ccagagaaga accgagttct gagagtctac agcaaaatta ctgggcacta
     3841 gaaaggaaga ctgggctccg aatcctctta gagccttgga catctctggc ccaaagcaat
     3901 ccaagtaagt acccagtcaa cgcgtgcggc cgcaggaacc cctagtgatg gagttggcca
     3961 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     4021 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggacatgtg agcaaaaggc
     4081 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4141 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4201 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4261 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4321 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4381 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4441 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4501 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4561 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4621 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgc
//
