LOCUS       sequence-470647-        5383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0891m 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 5383)
  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 5383)
  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: AiP13408 -
            pAAV-AiE0891m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(256..1047,1048..1116))
                     /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     879..898
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1117..1221)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1289..1307
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1345..1367)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1467..1486
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1503..1958)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1640..1661)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1852..1871
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2033..2173
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2044..2173
                     /label=AAV2 ITR
     polyA_signal    2291..2402
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2589..2609
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2696..3073)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(3104..3169)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(3182..3238)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(join(3248..3958,3959..3961,3962..3964))
                     /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(3290..3311)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3639..3658
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      3961..3970
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4689..4818
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4880..5383,1..85))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4960..4979)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
       61 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
      121 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
      181 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
      241 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
      301 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
      361 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
      421 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
      481 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
      541 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
      601 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
      661 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
      721 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
      781 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
      841 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
      901 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
      961 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     1021 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     1081 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     1141 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     1201 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     1261 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     1321 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     1381 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     1441 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     1501 taaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     1561 attttttaac caatagaccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga
     1621 gatagagttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     1681 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     1741 caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     1801 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     1861 agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     1921 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct ttgacgtatg
     1981 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg cccctgcagg
     2041 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
     2101 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     2161 actaggggtt cctgcggccg ctcggtccgc acgtgagatc tcatgcctgc tattgtcttc
     2221 ccaatcctcc cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact
     2281 cagacaatgc gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc
     2341 cagggtcaag gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac
     2401 agtcgaagat ctctcgagcc acggaattgt cagtgcccaa cagccgagcc cctgtccagc
     2461 agcgggcaag gcaggcggcg atgagttccg ccgtggcaag aactaaccag gatttataca
     2521 aggaggagaa aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag
     2581 cgtatccaca tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa
     2641 ttttgtaatc cagaggttga ttatgatatc gaattcgcgg ccgcggcgcg cctcattact
     2701 tagcgctggt gtacttggtg atggccttag taccctcgga cacggcgtgc ttggccaact
     2761 ccccaggcag cagcaggcgc acggccgtct ggatctcccg ggaggtgatg gtcgagcgct
     2821 tgttgtaatg cgccaggcgg gaagcctctc ctgcgatgcg ctcgaaaatg tcgttcacaa
     2881 acgaattcat gatgcccatg gccttggacg aaatgccggt gtcagggtgg acctgcttca
     2941 gaaccttgta cacatagatg gaatagctct ccttgcggct gcgcttgcgc ttcttgccgc
     3001 ctttcttctg cgccttagtc accgccttct tggagccctt tttcggggcg ggagcagact
     3061 tcgctggctc tgggcctccg gagcctccag atctgagtcc ggacttgtca tcgtcatcct
     3121 tgtaatcgat gtcatgatct ttataatctc cgtcatggtc tttgtagtct ccactagcac
     3181 caggccctgg attctcctct acatcccctg cctgtttcaa cagtgagaag ttggtagcac
     3241 cagagccctt gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca
     3301 gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagcttggac tggtagctca
     3361 ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt
     3421 ggtcggcgag ctgcacgccg ccgtcctcga tgttgtggcg gatcttgaag ttggccttga
     3481 tgccgttctt ctgcttgtcg gcggtgatat agacgttgtg gctgttgtag ttgtactcca
     3541 gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt
     3601 tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtctt
     3661 tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt
     3721 gctgcttcat gtggtcgggg tagcgggcga agcactgcac gccgtagccc agggtggtca
     3781 cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatcagcttc agggtcagct
     3841 tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgttgacat
     3901 cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttggaga
     3961 ccatggtggc gaccggtagc gctagcttcg aaagatctgg atcccagaag caaatgtaag
     4021 caatagatgg ctctgccctg acttttatgc ccagcccgag ctctgctctg ccattttggt
     4081 ccatggttac ctggccacta ggtataacat gacagttagt actgtgttgg tgcataacat
     4141 gacacagata ttcagcagag aagaggccta ccttctggca gctggaaaac aaggcgagga
     4201 gagaaagaga aaaggagaaa gagagaggga gagggagaga gagggagaga gagggagaga
     4261 gagaataacg caagcccaga atacccatat tcctttcaga gtacatgtca tttgacctca
     4321 tttccttcca ctaggctgtg ccccaaaagc ttctatctcc tccatttagc atcatgggcc
     4381 aagggccaag tgtttagtat atgtatgtct ggggtgatat ttcacattca tctaaatcat
     4441 aatatccagg gtatagctca ctggtaaatg acagcatacc agaccccaag ttttagcccc
     4501 agaattacaa aaactaccag taacaacagc aacagcaaaa tatggctcat agcaccaagt
     4561 cactactaac aaaagtaaat aaaaataaac tattaattat ctgtctttaa tgatagtaga
     4621 actcaaaatc ttaaaatata cttcaagaaa aagaacaagc tttactgcaa atgtacgcgt
     4681 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     4741 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag
     4801 cgcgcagctg cctgcaggac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4861 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4921 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4981 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5041 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5101 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5161 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5221 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5281 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5341 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctc
//
