LOCUS       sequence-466325-        4744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2541m 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 4744)
  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 4744)
  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: AiP1906 - pAAV-AiE2541m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     69..88
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(105..560)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(242..263)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     454..473
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   635..775
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   646..775
                     /label=AAV2 ITR
     polyA_signal    893..1004
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1191..1211
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1304..2014,2015..2017,2018..2020))
                     /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(1346..1367)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1695..1714
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1954..1975
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2017..2026
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2652..2781
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2843..3431)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2923..2942)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3602..4393,4394..4462))
                     /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     4225..4244
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4463..4567)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4635..4653
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4691..4713)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
       61 taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg taaacgttaa
      121 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaatagac
      181 cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagagt tgagtgttgt
      241 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
      301 aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg
      361 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
      421 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
      481 taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
      541 tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac
      601 agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg cgctcgctcg
      661 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
      721 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc
      781 cgctcggtcc gcacgtgaga tctcatgcct gctattgtct tcccaatcct cccccttgct
      841 gtcctgcccc accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat
      901 ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac
      961 gggggagggg caaacaacag atggctggca actagaaggc acagtcgaag atctctcgag
     1021 ccacggaatt gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg
     1081 cgatgagttc cgccgtggca agaactaacc aggatttata caaggaggag aaaatgaaag
     1141 ccatacggga agcaatagca tgatacaaag gcattaaagc agcgtatcca catagcgtaa
     1201 aaggagcaac atagttaaga ataccagtca atctttcaca aattttgtaa tccagaggtt
     1261 gattatgata tcgaattcgc ggccgcggcg cgccgtcgac ttacttgtac agctcgtcca
     1321 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     1381 cgttggggtc tttgctcagc ttggactggt agctcaggta gtggttgtcg ggcagcagca
     1441 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt
     1501 cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg
     1561 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     1621 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     1681 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     1741 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     1801 gggcgaagca ctgcacgccg tagcccaggg tggtcacgag ggtgggccag ggcacgggca
     1861 gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     1921 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     1981 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtagcgcta
     2041 gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat agatggctct gccctgactt
     2101 ttatgcccag cccgagctcc ggcctgccaa ggaggaagca gttagaccct tatcaagcat
     2161 cgctgagtga gatcatgtgt caaaactgag acggttagga ccaagaaagt gtggctagca
     2221 agaatgcatc tgaagggcac tgatgggaga gagaaatgaa agaggtaagg tttgcccaga
     2281 ttctcaccgt gagcaagaag ccaagcccac accacaaagt tgcctctgta tccagccagc
     2341 tgatgaatcc ttctctgaag acatggcggg ggtggggggg acactcctgg aaatgagtca
     2401 ggacaataat aagttggggc tttaaaaata actagacaag cagcaaaaag tgagacggca
     2461 cagtcccagg ctggaagcag cggatccgtc gctgtcaggg aggccgcaca ttgctcatct
     2521 ttgcgcaggt tcccttgact tcggagtgac attatttcct cagctcactt caggctccct
     2581 ctgaaatatt ccatgactaa catccagtcc agagattaca cgattattac tcttacaacg
     2641 cgtgcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     2701 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc
     2761 gagcgcgcag ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2821 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2881 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2941 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3001 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3061 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3121 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3181 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3241 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3301 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3361 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3421 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3481 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3541 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3601 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3661 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3721 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3781 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3841 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3901 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3961 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4021 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4081 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4141 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4201 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4261 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4321 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4381 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4441 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4501 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4561 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     4621 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     4681 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     4741 atgc
//
