LOCUS       sequence-466295-        5040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2411m 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 5040)
  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 5040)
  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: AiP1710 - pAAV-AiE2411m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   254..383
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(445..1033)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(525..544)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1204..1995,1996..2064))
                     /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     1827..1846
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2065..2169)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2237..2255
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2293..2315)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2415..2434
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2451..2906)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2588..2609)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2800..2819
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2981..3121
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2992..3121
                     /label=AAV2 ITR
     polyA_signal    3239..3350
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3537..3557
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3650..4360,4361..4363,4364..4366))
                     /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(3692..3713)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4041..4060
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4300..4321
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4363..4372
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tcaacacaac tgtcagtgct catcccgccc cctctcagca gcctctgacc cggttcaacc
       61 tgactcctgg gaactgactg acactctcct gcccacgtgc tgcttcacag gccacttggg
      121 tgagttcttg ttttgaggat cacaacttgt tcaagcctca agggttccga tctttggctt
      181 cttttcttta atgtgcatgt gtgcatatat gtaagtgtgt gtcgattatt actcttacaa
      241 cgcgtgcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
      301 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcggcct cagtgagcga
      361 gcgagcgcgc agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
      421 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
      481 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      541 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
      601 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
      661 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
      721 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
      781 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
      841 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
      901 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
      961 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1021 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1081 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     1141 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     1201 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     1261 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     1321 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     1381 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     1441 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     1501 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     1561 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     1621 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     1681 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     1741 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     1801 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     1861 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     1921 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     1981 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     2041 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     2101 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     2161 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     2221 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     2281 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     2341 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     2401 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata aaattgtaaa
     2461 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     2521 atagaccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga tagagttgag
     2581 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     2641 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt
     2701 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     2761 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     2821 gggcgctaag gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     2881 gcttaatgcg ccgctacagg gcgcgtacta tggttgcttt gacgtatgcg gtgtgaaata
     2941 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc cctgcaggca gctgcgcgct
     3001 cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg
     3061 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
     3121 tgcggccgct cggtccgcac gtgagatctc atgcctgcta ttgtcttccc aatcctcccc
     3181 cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga
     3241 tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga
     3301 aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaagatct
     3361 ctcgagccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc
     3421 aggcggcgat gagttccgcc gtggcaagaa ctaaccagga tttatacaag gaggagaaaa
     3481 tgaaagccat acgggaagca atagcatgat acaaaggcat taaagcagcg tatccacata
     3541 gcgtaaaagg agcaacatag ttaagaatac cagtcaatct ttcacaaatt ttgtaatcca
     3601 gaggttgatt atgatatcga attcgcggcc gcggcgcgcc gtcgacttac ttgtacagct
     3661 cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc
     3721 gcttctcgtt ggggtctttg ctcagcttgg actggtagct caggtagtgg ttgtcgggca
     3781 gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc
     3841 cgccgtcctc gatgttgtgg cggatcttga agttggcctt gatgccgttc ttctgcttgt
     3901 cggcggtgat atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt
     3961 tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct
     4021 cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct
     4081 cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg
     4141 ggtagcgggc gaagcactgc acgccgtagc ccagggtggt cacgagggtg ggccagggca
     4201 cgggcagctt gccggtggtg cagatcagct tcagggtcag cttgccgtag gtggcatcgc
     4261 cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca
     4321 ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggtg gcgaccggta
     4381 gcgctagctt cgaaagatct ggatcccaga agcaaatgta agcaatagat ggctctgccc
     4441 tgacttttat gcccagcccg agctccgaat ctaaaaatat atgttataca ttatataatt
     4501 atatagaatc tcaaatgaaa gttcgatgga taatgatcac ccggactttc ttccatgacg
     4561 ggcattaaca tttatgagca tctcctctga agtgccatgt ctcttcgatt cacaccaggg
     4621 atgtttcttc ttagatctcc aacatctctc ctcttccctc atttgtatag acagaaactt
     4681 gtgtagtctc tcaatgctgc atccacttcc atctgtgtct aaactctcag cttccctctt
     4741 gtcctcacca atgcttaagg ctccctctgc ctattcaaca cctctctagt ttgtatcccc
     4801 tatctcctgg atggtcccac cagctcctga acacagcaca gagtcacccg tatcaagcaa
     4861 gagaaccctc gatttaccta aactcccagc tcctccttcc taagagctct ctctattcgc
     4921 ttactgcatt tcctgctgtt tgctcttaaa cctgaacagg caagctctct ccccagtccc
     4981 acccactgca gttattctag ttggctcact gtcaagctac tagtatagtt tccaagctac
//
