LOCUS       sequence-470427-        4731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2154m 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 4731)
  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 4731)
  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: AiP1383 - pAAV-AiE2154m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4731
                     /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      743..752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(749..751,752..754,755..1465)
                     /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(794..815)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1055..1074)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1402..1423
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1558..1578)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1765..1876
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1994..2134
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1994..2123
                     /label=AAV2 ITR
     rep_origin      2209..2664
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2296..2315)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2506..2527
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2681..2700)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2800..2822
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2860..2878)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2946..3050
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3051..3119,3120..3911)
                     /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(3269..3288)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4082..4670
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4571..4590
                     /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 gcgttgtaag agtaataatc gccggcgcca gggcggtccc
      181 aggcctcaga gctccaaggg acgggaagta cccagtgaag caagggacag ccctggtgga
      241 ataactcctt cacagctcca tcccagaagt cagcggccac ccagggcagc tggtacacca
      301 cagtgacgaa cctgattgcc agtcagcctg gcagccacaa gaatggggta attctacaag
      361 gaccagggcc agagctgtct gtaaggggtt aaaaaaaaaa aaaaaaaaaa ggacactcct
      421 cggtccccag gagttaaagg gaatcctggc aagccaccca ggagaactct ctggaggcat
      481 gtttgacttt taaattccaa actctagctg ccccagtgct tcagagagat gttgccaaac
      541 gagttcaaag agacatgcat gggtcaaaca gtaaccactc tacaagagaa gctgctttgt
      601 gtgttctgag gtagctgata acatggacac aggaatactc tcgccggcgg agctcgggct
      661 gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct
      721 ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
      781 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
      841 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac
      901 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg
      961 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1021 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1081 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1141 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1201 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     1261 catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1321 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga
     1381 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1441 tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc gaattcgata
     1501 tcataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     1561 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     1621 ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga
     1681 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     1741 ttccgtggct cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     1801 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     1861 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     1921 ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg
     1981 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     2041 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     2101 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     2161 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     2221 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2281 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2341 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2401 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     2461 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2521 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     2581 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     2641 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     2701 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     2761 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     2821 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     2881 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     2941 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3001 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3061 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3121 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3181 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3241 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3301 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3361 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3421 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3481 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3541 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3601 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3661 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3721 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3781 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3841 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3901 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3961 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4021 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4081 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4141 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4201 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4261 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4321 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4381 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4441 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4501 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4561 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4621 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4681 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
