LOCUS       sequence-470676-        5397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0981h 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 5397)
  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 5397)
  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: AiP13464 -
            pAAV-AiE0981h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5397
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      47..56
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(53..55,56..58,59..769)
                     /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(359..378)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     706..727
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             779..835
                     /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             848..913
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             944..1321
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(1408..1428)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1615..1726
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1844..1984
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1844..1973
                     /label=AAV2 ITR
     rep_origin      2059..2514
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2146..2165)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2356..2377
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2531..2550)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2650..2672
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2710..2728)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2796..2900
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2901..2969,2970..3761)
                     /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(3119..3138)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3932..4520
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4421..4440
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4582..4711
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 catttgcttc tgggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtctc
       61 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgatgt
      121 caacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      181 gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
      241 caccctgggc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
      301 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaaaga
      361 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
      421 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
      481 gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
      541 ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta
      601 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
      661 ctaccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
      721 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gctctggtgc
      781 taccaacttc tcactgttga aacaggcagg ggatgtagag gagaatccag ggcctggtgc
      841 tagtggagac tacaaagacc atgacggaga ttataaagat catgacatcg attacaagga
      901 tgacgatgac aagtccggac tcagatctgg aggctccgga ggcccagagc cagcgaagtc
      961 tgctcccgcc ccgaaaaagg gctccaagaa ggcggtgact aaggcgcaga agaaaggcgg
     1021 caagaagcgc aagcgcagcc gcaaggagag ctattccatc tatgtgtaca aggttctgaa
     1081 gcaggtccac cctgacaccg gcatttcgtc caaggccatg ggcatcatga attcgtttgt
     1141 gaacgacatt ttcgagcgca tcgcaggaga ggcttcccgc ctggcgcatt acaacaagcg
     1201 ctcgaccatc acctcccggg agatccagac ggccgtgcgc ctgctgctgc ctggggagtt
     1261 ggccaagcac gccgtgtccg agggtactaa ggccatcacc aagtacacca gcgctaagta
     1321 atgaggcgcg ccgcggccgc gaattcgata tcataatcaa cctctggatt acaaaatttg
     1381 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1441 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     1501 taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     1561 gacaggggct cggctgttgg gcactgacaa ttccgtggct cgagagatct tcgactgtgc
     1621 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     1681 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     1741 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     1801 acaatagcag gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     1861 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     1921 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     1981 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     2041 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2101 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     2161 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2221 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     2281 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2341 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     2401 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     2461 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     2521 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     2581 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     2641 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     2701 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     2761 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2821 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2881 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2941 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3001 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3061 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3121 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3181 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3241 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3301 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3361 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3421 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     3481 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3541 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3601 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3661 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3721 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3781 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3841 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3901 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3961 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4021 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4081 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4141 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4201 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4261 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4321 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4381 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4441 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4501 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4561 gctggccttt tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg
     4621 cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag
     4681 ggagtggcca actccatcac taggggttcc tgcggccgca cgcgtcgagt gaacggatga
     4741 gtggaattta ctgcacccat aagtagaatc ttgagccctg tgcctgacct agactaggcc
     4801 ctcagacatg gttcagggct ggcaggatga gtaagtgcct actgaatgca aaggctgcag
     4861 aggcgagagg caggtggtgg ggatgcagct ccttgtgcac catctggcaa gggaagagaa
     4921 ggtgactctt taccaccaac ccctcccctg tgccaggctc ctgcggtaca caagacagga
     4981 gcatggagga aaatcaaacc aagcctccgc cagtgggatg gaaaaggttt ccaaataaac
     5041 cagcggccaa ggctcagaat ggctgaccag aggccttgcc cagaggacag agaaatccac
     5101 acggctgcct gatgaaatcg agaggacacg ccaactagat tagcacagat catcagccta
     5161 caaccaaagt acctgctcca ggagcacacg gagctttggc gcccaccttc aggctctggc
     5221 catgccagac atcatgaagg gcagcctcag gctccctcag aaaagcaaag aacaagaact
     5281 gcaagagttt cgtttaaaat ccatgttaag tctgaacagc ttcacaattg caattagcca
     5341 gacagagctc gagctcgggc tgggcataaa agtcagggca gagccatcta ttgctta
//
