LOCUS       sequence-470703-        5353 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0985h 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 5353)
  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 5353)
  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: AiP13494 -
            pAAV-AiE0985h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5353
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(148..736)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(228..247)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(907..1698,1699..1767))
                     /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     1530..1549
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1768..1872)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1940..1958
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1996..2018)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2118..2137
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2154..2609)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2291..2312)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2503..2522
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2684..2824
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2695..2824
                     /label=AAV2 ITR
     polyA_signal    2942..3053
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3240..3260
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3347..3724)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(3755..3820)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(3833..3889)
                     /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(3899..4609,4610..4612,4613..4615))
                     /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(3941..3962)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4290..4309
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      4612..4621
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   join(5310..5353,1..86)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag
       61 cgagcgagcg cgcagctgcc tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa
      121 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      181 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
      241 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
      301 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
      361 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
      421 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
      481 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
      541 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
      601 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
      661 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
      721 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
      781 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
      841 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
      901 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
      961 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1021 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1081 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1141 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1201 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1261 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     1321 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     1381 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     1441 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     1501 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     1561 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     1621 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     1681 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     1741 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     1801 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     1861 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     1921 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     1981 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     2041 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     2101 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc ataaaattgt
     2161 aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2221 ccaatagacc gaaatcggca aaatccctta taaatcaaaa gaatagcccg agatagagtt
     2281 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2341 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac ccaaatcaag
     2401 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     2461 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     2521 agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2581 cgcgcttaat gcgccgctac agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa
     2641 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc
     2701 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc
     2761 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
     2821 tcctgcggcc gctcggtccg cacgtgagat ctcatgcctg ctattgtctt cccaatcctc
     2881 ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg
     2941 cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa
     3001 ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaaga
     3061 tctctcgagc cacggaattg tcagtgccca acagccgagc ccctgtccag cagcgggcaa
     3121 ggcaggcggc gatgagttcc gccgtggcaa gaactaacca ggatttatac aaggaggaga
     3181 aaatgaaagc catacgggaa gcaatagcat gatacaaagg cattaaagca gcgtatccac
     3241 atagcgtaaa aggagcaaca tagttaagaa taccagtcaa tctttcacaa attttgtaat
     3301 ccagaggttg attatgatat cgaattcgcg gccgcggcgc gcctcattac ttagcgctgg
     3361 tgtacttggt gatggcctta gtaccctcgg acacggcgtg cttggccaac tccccaggca
     3421 gcagcaggcg cacggccgtc tggatctccc gggaggtgat ggtcgagcgc ttgttgtaat
     3481 gcgccaggcg ggaagcctct cctgcgatgc gctcgaaaat gtcgttcaca aacgaattca
     3541 tgatgcccat ggccttggac gaaatgccgg tgtcagggtg gacctgcttc agaaccttgt
     3601 acacatagat ggaatagctc tccttgcggc tgcgcttgcg cttcttgccg cctttcttct
     3661 gcgccttagt caccgccttc ttggagccct ttttcggggc gggagcagac ttcgctggct
     3721 ctgggcctcc ggagcctcca gatctgagtc cggacttgtc atcgtcatcc ttgtaatcga
     3781 tgtcatgatc tttataatct ccgtcatggt ctttgtagtc tccactagca ccaggccctg
     3841 gattctcctc tacatcccct gcctgtttca acagtgagaa gttggtagca ccagagccct
     3901 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
     3961 tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga ctggtagctc aggtagtggt
     4021 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
     4081 gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa gttggccttg atgccgttct
     4141 tctgcttgtc ggcggtgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc
     4201 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
     4261 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtct ttgaagaaga
     4321 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     4381 tgtggtcggg gtagcgggcg aagcactgca cgccgtagcc cagggtggtc acgagggtgg
     4441 gccagggcac gggcagcttg ccggtggtgc agatcagctt cagggtcagc ttgccgtagg
     4501 tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgttgaca tcgccgtcca
     4561 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttggag accatggtgg
     4621 cgaccggtag cgctagcttc gaaagatctg gatcccagaa gcaaatgtaa gcaatagatg
     4681 gctctgccct gacttttatg cccagcccga gctctggact cacactcccg atttcagaat
     4741 gcactctccc atcaacttca tgttctccac caccggcctg ctctctgaga gctctgcaaa
     4801 gtaaggtttc tcaatttgca ctgctgacat tctgggttgg ataatttttg ttgtgggagc
     4861 tgccctgttc attatgggat gcaagcagca tccctggctc atacccacca gagcacaaac
     4921 tcctagctgt gaaaaccaaa gatgtctcag acactgccag atgtctccaa gaggaacaaa
     4981 gtcattccca gctgggaacc actgctctta aagggattgg ttctcaacct tctccagcag
     5041 actttgccat cagactgctg gtgtagttac caaccaggtg ctcccagaaa tgtatccttg
     5101 gtggagtgag gaaggcagga acctcatagc accgaggttt ggtcctgcct tactctgaat
     5161 aaggaaccag tgtgtcactg actcaaaaac tgactagctg caggacagag aaccagacaa
     5221 cctgagagaa gaagatgact acatgtttct tagattgctt ctatctcatt tatagctaga
     5281 gggaagactg gagctacgcg tgcggccgca ggaaccccta gtgatggagt tggccactcc
     5341 ctctctgcgc gct
//
