LOCUS       sequence-470730-        5157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0588h 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 5157)
  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 5157)
  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: AiP13656 -
            pAAV-AiE0588h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      16..604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     505..524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   666..795
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1288..1297
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1294..1296,1297..1299,1300..2010)
                     /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(1600..1619)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1947..1968
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2020..2076
                     /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             2089..2154
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2185..2562
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2649..2669)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2856..2967
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3085..3225
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3085..3214
                     /label=AAV2 ITR
     rep_origin      3300..3755
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3387..3406)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3597..3618
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3772..3791)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3891..3913
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3951..3969)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4037..4141
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4142..4210,4211..5002)
                     /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(4360..4379)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
       61 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
      121 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
      181 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
      241 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
      301 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
      361 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      421 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
      481 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
      541 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
      601 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
      661 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
      721 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      781 tcactagggg ttcctgcggc cgcacgcgtc catatagggg cttagatctt ttttagacct
      841 aaattgctgt gttttcccac catggttcat ttttcttagt gatagaaaca aaatcttttt
      901 cttgggcaca atttaaactt gtaattttgg aactacagct atggatatga agctcttaat
      961 catcaaggca tttttcctaa acacttaagc tgatgcaatt ttcagaaatt cagaaaacaa
     1021 attttgcaat cacttttccc agtgattaaa aagaaggtaa aaggacagaa atgagaggca
     1081 taattatatg gtacatttag tcaacaactg tacaggcaaa atgtcaaaaa aataaggcac
     1141 ccatcaagca tttcaaccgt acctgactaa ccatgttatg gatttgcctc ctgggagctc
     1201 gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt ctgggatcca
     1261 gatctttcga agctagcgct accggtcgcc accatggtct ccaagggcga ggagctgttc
     1321 accggggtgg tgcccatcct ggtcgagctg gacggcgatg tcaacggcca caagttcagc
     1381 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc
     1441 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcgtg
     1501 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1561 cccgaaggct acgtccagga gcgcaccatc ttcttcaaag acgacggcaa ctacaagacc
     1621 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1681 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1741 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     1801 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc
     1861 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc
     1921 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1981 atcactctcg gcatggacga gctgtacaag ggctctggtg ctaccaactt ctcactgttg
     2041 aaacaggcag gggatgtaga ggagaatcca gggcctggtg ctagtggaga ctacaaagac
     2101 catgacggag attataaaga tcatgacatc gattacaagg atgacgatga caagtccgga
     2161 ctcagatctg gaggctccgg aggcccagag ccagcgaagt ctgctcccgc cccgaaaaag
     2221 ggctccaaga aggcggtgac taaggcgcag aagaaaggcg gcaagaagcg caagcgcagc
     2281 cgcaaggaga gctattccat ctatgtgtac aaggttctga agcaggtcca ccctgacacc
     2341 ggcatttcgt ccaaggccat gggcatcatg aattcgtttg tgaacgacat tttcgagcgc
     2401 atcgcaggag aggcttcccg cctggcgcat tacaacaagc gctcgaccat cacctcccgg
     2461 gagatccaga cggccgtgcg cctgctgctg cctggggagt tggccaagca cgccgtgtcc
     2521 gagggtacta aggccatcac caagtacacc agcgctaagt aatgaggcgc gccgcggccg
     2581 cgaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2641 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2701 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt
     2761 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     2821 ggcactgaca attccgtggc tcgagagatc ttcgactgtg ccttctagtt gccagccatc
     2881 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2941 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3001 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat
     3061 ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     3121 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     3181 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     3241 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     3301 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3361 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3421 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3481 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     3541 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3601 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     3661 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3721 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     3781 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     3841 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     3901 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     3961 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4021 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4081 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4141 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4201 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4261 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4321 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4381 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4441 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4501 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4561 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4621 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4681 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     4741 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4801 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4861 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4921 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4981 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5041 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5101 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaa
//
