LOCUS       sequence-460622-        4618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0465m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific brain cell types.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4618)
  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 4618)
  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: AiP12425 - pAAV-AiE0465m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      630..639
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(636..638,639..641,642..1352)
                     /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(681..702)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(942..961)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1289..1310
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1445..1465)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1652..1763
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1881..2021
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1881..2010
                     /label=AAV2 ITR
     rep_origin      2096..2551
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2183..2202)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2393..2414
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2568..2587)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2687..2709
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2747..2765)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2833..2937
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2938..3006,3007..3798)
                     /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(3156..3175)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3969..4557
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4458..4477
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtactgac cgtttgcaca gtgcctatgt tcatttataa
      181 taaacctgcg tgtcacgaat tccaacccaa agtgtgtgcg aataatggaa aatataagaa
      241 aaatctactt aagcatctgg caggaaaata ccaataatta tggtgtttca agtcaatttg
      301 tttctttttc aagattttct ttgaaagtat acatgaaacc tgtaaaaagg aaactgattg
      361 cttcctcagg gtaaacagct ctcgatgata gaagtcattt acttttgatt tccttttagg
      421 tgagtattca gaacgatact caaatgtgaa acaaatttaa tcaaggggaa aaaggtgcaa
      481 aaatactatg gaaaatgtca atcactagtt ttatggagaa gctgaaaatt cgtcctgagc
      541 tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc ttctgggatc
      601 cagatctttc gaagctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt
      661 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
      721 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct
      781 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcg
      841 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
      901 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
      961 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     1021 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     1081 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
     1141 gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
     1201 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga
     1261 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     1321 ggatcactct cggcatggac gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa
     1381 ttcgatatca taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     1441 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     1501 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca
     1561 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     1621 ctgacaattc cgtggctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt
     1681 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     1741 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     1801 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca
     1861 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     1921 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     1981 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     2041 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     2101 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2161 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2221 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2281 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     2341 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2401 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     2461 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2521 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     2581 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     2641 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     2701 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     2761 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     2821 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2881 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2941 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3001 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3061 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3121 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3181 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3241 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3301 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3361 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3421 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3481 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3541 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3601 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3661 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3721 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3781 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3841 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3901 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3961 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4021 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4081 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4141 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4201 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4261 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4321 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4381 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4441 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4501 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4561 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
