LOCUS       sequence-460615-        4516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0725m 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 4516)
  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 4516)
  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: AiP12864 - pAAV-AiE0725m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4516
                     /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      528..537
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(534..536,537..539,540..1250)
                     /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(579..600)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(840..859)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1187..1208
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1343..1363)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1550..1661
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1779..1919
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1779..1908
                     /label=AAV2 ITR
     rep_origin      1994..2449
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2081..2100)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2291..2312
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2466..2485)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2585..2607
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2645..2663)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2731..2835
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2836..2904,2905..3696)
                     /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(3054..3073)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3867..4455
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4356..4375
                     /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 gcgtttttgt gtgtctgtcc cccagtgccg ggctctgccc
      181 ctcctctgtc tttccctgcc tgctctgtgc atttactctg aactcgagca ggcctatttc
      241 caaacaccgt ctagcgcggg gctgtgccag ctccggctct gtcaggacca acttcccaaa
      301 gtgttcgcag attctcagag cttagctctt ctcagagttt gccatttttt tttaaaaaga
      361 gatttatttt ttattttaaa tgatagacgc tcgtgggtgt ctgtgtgggg gattgtacac
      421 atgtgtgggg gtgtgagctc gggctgggca taaaagtcag ggcagagcca tctattgctt
      481 acatttgctt ctgggatcca gatctttcga agctagcgct accggtcgcc accatggtga
      541 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      601 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      661 tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga
      721 ccaccctggg ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
      781 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
      841 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
      901 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
      961 agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca
     1021 aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact
     1081 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1141 gctaccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1201 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taagtcgacg
     1261 gcgcgccgcg gccgcgaatt cgatatcata atcaacctct ggattacaaa atttgtgaaa
     1321 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     1381 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     1441 cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     1501 gggctcggct gttgggcact gacaattccg tggctcgaga gatcttcgac tgtgccttct
     1561 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     1621 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     1681 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     1741 agcaggcatg agatctcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt
     1801 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     1861 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     1921 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     1981 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2041 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     2101 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2161 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     2221 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2281 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     2341 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     2401 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     2461 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     2521 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2581 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     2641 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     2701 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2761 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2821 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2881 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     2941 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3001 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3061 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3121 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3181 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3241 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3301 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3361 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3421 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3481 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3541 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3601 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3661 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3721 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3781 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3841 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3901 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3961 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4021 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4081 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4141 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4201 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4261 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4321 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4381 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4441 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4501 ccttttgctc acatgt
//
