LOCUS       sequence-470557-        4827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0586m 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 4827)
  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 4827)
  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: AiP12875 - pAAV-AiE0586m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   148..277
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(339..927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(419..438)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1098..1889,1890..1958))
                     /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     1721..1740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1959..2063)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2131..2149
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2187..2209)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2309..2328
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2345..2800)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2482..2503)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2694..2713
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2875..3015
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2886..3015
                     /label=AAV2 ITR
     polyA_signal    3133..3244
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3431..3451
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3544..4254,4255..4257,4258..4260))
                     /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(3586..3607)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3935..3954
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4194..4215
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4257..4266
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tcctagtgca cacactattt actatttgcc tctttctctt ccatctctcc accacttact
       61 tgttcatcta agtagtaccc agggcctctg ggtgtctggg gtcaccatag gagtgttatg
      121 ccctggctat gctacgcgtg cggccgcagg aacccctagt gatggagttg gccactccct
      181 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg
      241 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa
      301 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
      361 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      421 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
      481 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
      541 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
      601 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      661 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
      721 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
      781 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
      841 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
      901 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
      961 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     1021 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     1081 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     1141 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     1201 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     1261 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     1321 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     1381 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     1441 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     1501 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     1561 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     1621 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     1681 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     1741 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     1801 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     1861 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     1921 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     1981 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     2041 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     2101 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     2161 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     2221 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     2281 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     2341 cataaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     2401 tcatttttta accaatagac cgaaatcggc aaaatccctt ataaatcaaa agaatagccc
     2461 gagatagagt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     2521 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     2581 cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     2641 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     2701 aaagcgaaag gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     2761 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgta
     2821 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca
     2881 ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga
     2941 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
     3001 tcactagggg ttcctgcggc cgctcggtcc gcacgtgaga tctcatgcct gctattgtct
     3061 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
     3121 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
     3181 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
     3241 acagtcgaag atctctcgag ccacggaatt gtcagtgccc aacagccgag cccctgtcca
     3301 gcagcgggca aggcaggcgg cgatgagttc cgccgtggca agaactaacc aggatttata
     3361 caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc
     3421 agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca
     3481 aattttgtaa tccagaggtt gattatgata tcgaattcgc ggccgcggcg cgccgtcgac
     3541 ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag
     3601 gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggt agctcaggta
     3661 gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc
     3721 ggcgagctgc acgccgccgt cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc
     3781 gttcttctgc ttgtcggcgg tgatatagac gttgtggctg ttgtagttgt actccagctt
     3841 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     3901 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     3961 gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     4021 cttcatgtgg tcggggtagc gggcgaagca ctgcacgccg tagcccaggg tggtcacgag
     4081 ggtgggccag ggcacgggca gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc
     4141 gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     4201 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
     4261 ggtggcgacc ggtagcgcta gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat
     4321 agatggctct gccctgactt ttatgcccag cccgagctct cgcttctgat acctggggaa
     4381 aaaaaagggt tagtcatatg tcaaaaattt atctgaaaat gtctgccctc ttcacagact
     4441 ctagcagttc atagactgtg tagatgttgt ggtgggccaa ctcaaagccc tggacctggg
     4501 cctgggtggc agctgagcta gtttgcttgc cagcccgtac aatgatagca aactcttcag
     4561 cgctgccctg gcgaagctag ctcacccagt gctacagcca gcaaggggca gggccacatc
     4621 atttgtacaa tgccaggtgg tcaccaaggc caggagcatc cacctggcct ttggtggtga
     4681 gagacacccg ctcttttagt gccaccaaca cagaaatgga gctcagtaac agcacaggcc
     4741 aggacatcac tgtggcctca tgtggcattg ccagctattc atatgaggct gttttcacta
     4801 ccgtcaagtt tccagttcta tcccttt
//
