LOCUS       sequence-470526-        4886 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0561m 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 4886)
  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 4886)
  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: AiP12832 - pAAV-AiE0561m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4886
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(60..78)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        146..250
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(251..319,320..1111)
                     /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(469..488)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1282..1870
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1771..1790
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1932..2061
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2829..2838
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2835..2837,2838..2840,2841..3551)
                     /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(2880..2901)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3141..3160)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3488..3509
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3644..3664)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3851..3962
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4080..4220
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4080..4209
                     /label=AAV2 ITR
     rep_origin      4295..4750
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4382..4401)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4592..4613
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4767..4786)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4886,1..22)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
       61 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
      121 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
      181 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      241 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      301 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      361 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      421 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      481 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      541 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      601 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      661 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
      721 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
      781 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
      841 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
      901 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
      961 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     1021 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     1081 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1141 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1201 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1261 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1321 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1381 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1441 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1501 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1561 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1621 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1681 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1741 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1801 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1861 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1921 tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc
     1981 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
     2041 actccatcac taggggttcc tgcggccgca cgcgtgcaat tgacactttg tagaaacgag
     2101 gaattatact ctaagcacaa cacaagtttt ctgcatttgt tcctgaagaa agtccttatt
     2161 ttctaaggta ctcctaggcc ctgtctataa cattataatc atcttctagt tcatcacagc
     2221 taagtaaaga ttaaagcatt gcacatccag ctaagtactt ttcttttttt aaaaaagagt
     2281 tcatttcctc tctcttgaac ttgtttgcta taaatgtata taaatcaggc atgatgacaa
     2341 tttaataaga acttcaagac ctgttcatcc catgggatca ctactgtgac gtctttcggg
     2401 gagccttctg attgctcagc ccgaagcaca agccctataa aatgagagac tttgagatgc
     2461 tggtcagctt ggacagcaga gcactagcca gctacagcca accgttcccc aggaaccggg
     2521 aacagactgg tggagccttc cctagagcag aacttcagca ccctagacag ctgccacgaa
     2581 gccggaaagg cagccctgcc tgaaggaaac agccaagggt aagcaaaatc ttccacttct
     2641 tcctctcctc tctgtcactg ggactttgaa taggagtttg tgaaaatttc tctcttaaaa
     2701 agaattttac agtgttcgtc tttttgcaat agccagagct cgggctgggc ataaaagtca
     2761 gggcagagcc atctattgct tacatttgct tctgggatcc agatctttcg aagctagcgc
     2821 taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2881 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     2941 gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg
     3001 tgccctggcc caccctcgtg accaccctgg gctacggcgt gcagtgcttc gcccgctacc
     3061 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     3121 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     3181 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     3241 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg
     3301 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg
     3361 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     3421 tgcccgacaa ccactacctg agctaccagt ccaagctgag caaagacccc aacgagaagc
     3481 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     3541 agctgtacaa gtaagtcgac ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc
     3601 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     3661 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     3721 ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct
     3781 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggctcgag
     3841 agatcttcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3901 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3961 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     4021 ggggaggatt gggaagacaa tagcaggcat gagatctcac gtgcggaccg agcggccgca
     4081 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     4141 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     4201 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4261 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     4321 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     4381 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4441 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4501 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4561 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4621 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     4681 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4741 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4801 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4861 gcttacagac aagctgtgac cgtctc
//
