LOCUS       sequence-466287-        4928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2354m 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 4928)
  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 4928)
  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: AiP1675 - pAAV-AiE2354m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(512..1303,1304..1372))
                     /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     1135..1154
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1373..1477)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1545..1563
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1601..1623)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1723..1742
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1759..2214)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1896..1917)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2108..2127
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2289..2429
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2300..2429
                     /label=AAV2 ITR
     polyA_signal    2547..2658
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2845..2865
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2958..3668,3669..3671,3672..3674))
                     /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(3000..3021)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3349..3368
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3608..3629
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3671..3680
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4490..4619
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4681..4928,1..341))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4761..4780)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
       61 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
      121 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
      181 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
      241 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
      301 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
      361 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
      421 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
      481 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
      541 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
      601 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
      661 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
      721 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
      781 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
      841 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
      901 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
      961 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     1021 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     1081 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     1141 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     1201 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     1261 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     1321 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     1381 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     1441 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     1501 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     1561 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     1621 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     1681 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     1741 tactgagagt gcaccataaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt
     1801 tttgttaaat cagctcattt tttaaccaat agaccgaaat cggcaaaatc ccttataaat
     1861 caaaagaata gcccgagata gagttgagtg ttgttccagt ttggaacaag agtccactat
     1921 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     1981 tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2041 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     2101 gaaaggaagg gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca
     2161 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg
     2221 gttgctttga cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     2281 caggcgcccc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
     2341 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
     2401 agtggccaac tccatcacta ggggttcctg cggccgctcg gtccgcacgt gagatctcat
     2461 gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa
     2521 tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag
     2581 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
     2641 ggcaactaga aggcacagtc gaagatctct cgagccacgg aattgtcagt gcccaacagc
     2701 cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt ggcaagaact
     2761 aaccaggatt tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac
     2821 aaaggcatta aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca
     2881 gtcaatcttt cacaaatttt gtaatccaga ggttgattat gatatcgaat tcgcggccgc
     2941 ggcgcgccgt cgacttactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc
     3001 acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagcttggac
     3061 tggtagctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc
     3121 tgctggtagt ggtcggcgag ctgcacgccg ccgtcctcga tgttgtggcg gatcttgaag
     3181 ttggccttga tgccgttctt ctgcttgtcg gcggtgatat agacgttgtg gctgttgtag
     3241 ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc
     3301 tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg
     3361 ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg
     3421 aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga agcactgcac gccgtagccc
     3481 agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatcagcttc
     3541 agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg
     3601 ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg
     3661 cccttgctca ccatggtggc gaccggtagc gctagcttcg aaagatctgg atcccagaag
     3721 caaatgtaag caatagatgg ctctgccctg acttttatgc ccagcccgag ctccgggaag
     3781 gtggggaggt ggggagtggg ttggagggga agagaagaaa cactggtctg agtgagcagg
     3841 aggggagaga aggatgcctg gaggagatga gcttcaagca acaccatacc taaaggctta
     3901 aaatgtgagg gcctaagctc acagtgaata ggaaaagata aaagcatgtg gcagacacaa
     3961 tgcttatgtg caagctgcag gctctgttgg gacctggtgg ttgagctgtg ggttcctcct
     4021 tagtcccggt gagtatatgg ctggggagga agactgggct cccagggtcc gcggggaggc
     4081 tgcccctcac agtactccca tgcatttgcc tttgggtttt tctcttcctc ttccaggaaa
     4141 ccccccagcc attcatggag aaggatcagt tgctcctggc caggaagctg gaagaactga
     4201 aggcaaggct ggccctgggc accagagagg ctagcatacc accactgaat cccaaagccc
     4261 aggaagaagg taagtgccca ggaacaggca ggtagggcca taggacattg tagggctgta
     4321 gaacaccgtc ctttgaacca gaagcacagt agccagtgga gaagtaaaga cccaaggagg
     4381 ctgtctctaa gcaaaacctg ggctgagcct tctgacccat tcccttctga caccctcacc
     4441 ccagacacat tctctctccg attattactc ttacaacgcg tgcggccgca ggaaccccta
     4501 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     4561 aaggtcgccc gacgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcagga
     4621 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4681 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4741 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4801 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4861 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4921 caagctgg
//
