LOCUS       sequence-470706-        5912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0986h 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 5912)
  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 5912)
  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: AiP13495 -
            pAAV-AiE0986h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      472..1060
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     961..980
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1122..1251
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2499..2508
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2505..2507,2508..2510,2511..3221)
                     /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(2811..2830)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3158..3179
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3231..3287
                     /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             3300..3365
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3396..3773
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(3860..3880)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4067..4178
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4296..4436
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4296..4425
                     /label=AAV2 ITR
     rep_origin      4511..4966
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4598..4617)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4808..4829
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4983..5002)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5102..5124
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5162..5180)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5248..5352
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5353..5421,5422..5912,1..301)
                     /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(5571..5590)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
       61 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
      121 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
      181 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
      241 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
      301 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
      361 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
      421 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
      481 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
      541 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
      601 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
      661 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
      721 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
      781 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
      841 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
      901 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
      961 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     1021 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     1081 tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc
     1141 tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     1201 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca
     1261 cgcgtagact gtcacaagct agaactacct atggaacaat gtctcaaata ggtgaatgtg
     1321 tgtaggcttt gcgtctggta accatggtta ccatgggcac agggagagaa gcttcacact
     1381 aaagagtgag cagtaaataa cggtggaaga gcctcctccc gttgtctaca gggagtctgg
     1441 gcagggggct gctgggatac aatgcaggga ttgtgtctca agactgtaaa tgaaagacac
     1501 aattccttgc ttgcggcctt ggtgttcttt gctactgcgg gcctcttgta tggctaacca
     1561 gttgatcatt gtcaagaaat ggaatggctc atagacacgc actgcctcct cagatctcct
     1621 ggtggtccct gctgcaggag agaagacctc aggactgtga gaaagcggga ccacttctct
     1681 tagcatgagg accttaaaga actggtgtag gttgttagca gttgtggcag cctgctgcct
     1741 tggttatgac atctggccct tgctgaaatg gagaaggttg agcccatccc caggagttag
     1801 gatgcacagg tggggaagct cccagaactc cttgcctggg agggaaggca ggtgcataga
     1861 gtcttggctt agcccccgag tctctctgtg tctggcagct ctcactgagg accacacaca
     1921 ggccccagat gctccacctg ggccttgatc ctgtgctcag ctccacctcc ctttacagca
     1981 gatgccacct gctcctgctt ttatgatcta gctaccaatt tcccatttcc cttctgcgtg
     2041 caagctgtgg aaaatctctc aaagaagata ctagaccttt tgttctagga aagaatttat
     2101 atatgcttaa aataaaattc cttttactct gatggtaaaa gttaacctgg gccgctgtcc
     2161 cagactacct tgagtccttt tagtttttaa attttttttt tcccattcat tcaggaagca
     2221 tttaatgaat gtctcctgga taccgagcac tgtgacgtgg aatcaacaag aaaggtccgt
     2281 gacttcagga agctgagata gctcacaatc tagccagaga attggagata aacagacaat
     2341 tttaatataa tatgaaagaa ctaggatacg tgtgtgttca gggtggtcta ggagcacaga
     2401 ggaaagagct cgggctgggc ataaaagtca gggcagagcc atctattgct tacatttgct
     2461 tctgggatcc agatctttcg aagctagcgc taccggtcgc caccatggtc tccaagggcg
     2521 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgat gtcaacggcc
     2581 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2641 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
     2701 gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     2761 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaaa gacgacggca
     2821 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2881 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2941 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     3001 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     3061 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     3121 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     3181 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggctctggt gctaccaact
     3241 tctcactgtt gaaacaggca ggggatgtag aggagaatcc agggcctggt gctagtggag
     3301 actacaaaga ccatgacgga gattataaag atcatgacat cgattacaag gatgacgatg
     3361 acaagtccgg actcagatct ggaggctccg gaggcccaga gccagcgaag tctgctcccg
     3421 ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca gaagaaaggc ggcaagaagc
     3481 gcaagcgcag ccgcaaggag agctattcca tctatgtgta caaggttctg aagcaggtcc
     3541 accctgacac cggcatttcg tccaaggcca tgggcatcat gaattcgttt gtgaacgaca
     3601 ttttcgagcg catcgcagga gaggcttccc gcctggcgca ttacaacaag cgctcgacca
     3661 tcacctcccg ggagatccag acggccgtgc gcctgctgct gcctggggag ttggccaagc
     3721 acgccgtgtc cgagggtact aaggccatca ccaagtacac cagcgctaag taatgaggcg
     3781 cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat
     3841 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3901 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3961 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     4021 ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt
     4081 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     4141 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     4201 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     4261 aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     4321 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     4381 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     4441 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     4501 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4561 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     4621 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4681 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     4741 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4801 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     4861 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     4921 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     4981 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5041 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5101 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5161 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5221 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5281 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5341 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5401 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5461 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5521 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5581 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5641 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5701 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5761 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5821 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5881 gacaccacga tgcctgtagc aatggcaaca ac
//
