LOCUS       sequence-466331-        4869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2553m 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 4869)
  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 4869)
  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: AiP1925 - pAAV-AiE2553m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      457..466
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(463..465,466..468,469..1179)
                     /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(508..529)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(769..788)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1116..1137
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1272..1292)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1479..1590
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1708..1848
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1708..1837
                     /label=AAV2 ITR
     rep_origin      1923..2378
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2010..2029)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2220..2241
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2395..2414)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2514..2536
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2574..2592)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2660..2764
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2765..2833,2834..3625)
                     /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     complement(2983..3002)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3796..4384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4285..4304
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4446..4575
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ttttgctgct gatagtaggc tgtgagagca tttgctgcat tcgatgctgc agcagagagt
       61 gcagtgactc actaactggc actagagttt gctgttttca ttccactagt aaagcaatcc
      121 aaattgcctt acatgatttc aagagtaggg atgtctgacc actaccatgt gcccaaaggt
      181 aagggccacg gccattgggc aagcagcagt agttgtgttg tggagagatg attggaatgg
      241 tttttgtcgc tcatggaact aaattatgtt agagtgactg tgacaccaga gactgtttta
      301 ctcagctttt caccgttcct agcgtgggcc aatttaagca aaccgtgaaa taagcctttg
      361 acggagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc
      421 tgggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtgag caagggcgag
      481 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      541 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      601 ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc
      661 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
      721 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
      781 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
      841 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
      901 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
      961 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
     1021 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     1081 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1141 gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg
     1201 ccgcgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     1261 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     1321 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt
     1381 cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     1441 ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc
     1501 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     1561 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1621 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga
     1681 gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     1741 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     1801 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     1861 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     1921 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1981 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2041 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2101 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     2161 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2221 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     2281 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     2341 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     2401 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     2461 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     2521 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     2581 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     2641 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2701 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2761 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2821 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2881 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2941 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3001 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3061 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3121 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3181 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3241 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3301 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3361 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3421 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3481 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3541 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3601 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3661 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3721 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3781 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3841 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3901 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3961 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4021 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4081 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4141 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4201 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4261 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4321 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4381 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4441 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
     4501 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
     4561 tcactagggg ttcctgcggc cgcagccctg ggccttcctc agtcatgagg agtggacagt
     4621 gcttgctcag ttagagaaga tctctccact tcaattgacc tcgtagcagt ctcacaaagg
     4681 caaacctaga agctaactaa tgggcatgtg cacgcctttg tggagcaata gtgtctctcc
     4741 aggcaattac taatgcttgt caggttggca atcaaagcta accatcacaa atgtcaacgt
     4801 caaggtattt tggtttgttc caacagttta tttttggagt aggttgtgaa tcctgttcag
     4861 atggttaag
//
