LOCUS       sequence-432546-        5017 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2351m 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 5017)
  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 5017)
  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: AiP1671 - pAAV-AiE2351m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5017
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      532..541
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(538..540,541..543,544..1254)
                     /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(583..604)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(844..863)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1191..1212
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1347..1367)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1554..1665
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1783..1923
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1783..1912
                     /label=AAV2 ITR
     rep_origin      1998..2453
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2085..2104)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2295..2316
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2470..2489)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2589..2611
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2649..2667)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2735..2839
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2840..2908,2909..3700)
                     /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(3058..3077)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3871..4459
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4360..4379
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4521..4650
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 aatacataga ccacaaaagc accttttttt aaaattttga tttaaggaaa atagctgccc
       61 caaaatatac caaatcacaa gttgacaccc caatatgcca agaatttctg caaagcaaag
      121 gaagtaagta gacagtttac caaaaggaaa aatggataaa atgtatttta aaagactttt
      181 taaaatagta ttccaagaaa tgcaaactga agttaagatt tcttgtaatt ttattactaa
      241 acattgtaag tgtgtgctcc aagtgccatg actagcactg tgaggtaggt aagcctgcag
      301 cctaagatgg aaagcgatct cacccccttt ctactctgtc ctgtagtcac ttactaaatt
      361 cctttgactc tgtagttcca cggctgaaga cattatcctc agggcacagg ctggcagata
      421 aaagagccac agggctcgga gctcgggctg ggcataaaag tcagggcaga gccatctatt
      481 gcttacattt gcttctggga tccagatctt tcgaagctag cgctaccggt cgccaccatg
      541 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      601 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      661 aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      721 gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
      781 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      841 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      901 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
      961 ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
     1021 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
     1081 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1141 ctgagctacc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1201 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagtc
     1261 gacggcgcgc cgcggccgcg aattcgatat cataatcaac ctctggatta caaaatttgt
     1321 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     1381 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     1441 aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     1501 acaggggctc ggctgttggg cactgacaat tccgtggctc gagagatctt cgactgtgcc
     1561 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1621 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     1681 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     1741 caatagcagg catgagatct cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     1801 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     1861 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     1921 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     1981 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2041 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     2101 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2161 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     2221 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2281 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     2341 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     2401 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     2461 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     2521 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     2581 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     2641 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     2701 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2761 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2821 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2881 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2941 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3001 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3061 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3121 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3181 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3241 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3301 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3361 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3421 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3481 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3541 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3601 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3661 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3721 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3781 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3841 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3901 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3961 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4021 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4081 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4141 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4201 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4261 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4321 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4381 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4441 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4501 ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
     4561 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     4621 gagtggccaa ctccatcact aggggttcct gcggccgcac gcgttgtaag agtaataatc
     4681 gagtaataaa tgtttctatg aaagttaccc actttggtgt ggtggcatat gactgtgatc
     4741 ccaccacttg ggaggcagag aaaaaagaat tgtgagtctg aagacagttt ggattatata
     4801 agttagactc tatcctcaaa atgcctaggg gaaaggaaag aacatacaga attagtcttt
     4861 gtctcaaaca atgtcgctgt gtacttacat aaactagagt ttaataaaaa cttagaagaa
     4921 aacagggtga agaagtgata taatttggtt tctgtcatat aaaagctaca gaaaatggtg
     4981 cagagaaaga gaccagcaaa cttagtttta aaaagta
//
