LOCUS       sequence-461882-        5359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0984h 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 5359)
  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 5359)
  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: AiP13493 -
            pAAV-AiE0984h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      825..834
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(831..833,834..836,837..1547)
                     /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(1137..1156)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1484..1505
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1557..1613
                     /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             1626..1691
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1722..2099
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2186..2206)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2393..2504
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2622..2762
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2622..2751
                     /label=AAV2 ITR
     rep_origin      2837..3292
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2924..2943)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3134..3155
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3309..3328)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3428..3450
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3488..3506)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3574..3678
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3679..3747,3748..4539)
                     /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(3897..3916)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4710..5298
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5199..5218
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcaggac ggtgctctgc atagcaggat gaccagcatc
      181 cctgaaccca cctattaaat gccagcaaca cttccaatca ttgtgaaagt aaaatcactg
      241 cctcgccaaa actctcaaaa tgcttccctg ggggcagtag cactccctct gagtgagtac
      301 caccgtcaga gtgcctcgtc caaagcgctg tcttgattca tttcctgagg catatggcag
      361 tgcacctgcc gcacctcggc ttggacatgt gcactggaaa actggcaagg ggaggtgttc
      421 agacaaattg tcagacttgc tgctggcttc aggttagatg aggcgctgag cctgagtgtc
      481 tcacgctctc aaggaaagca ttttttgcat cactgaactg atgcaagaaa agaggtggaa
      541 agaggaggaa agcattctca tggtttgggc ctcctcagtc agccttacct gctcagggcc
      601 ccacagcact gggtagaaga ggtaacgacc aagtggattt tagccacagt gatagtgagc
      661 attagggaca caggccatca cccccaaaag ctcttttctt tcattgccta aaggagagca
      721 aggagtgtgg tgagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca
      781 tttgcttctg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc atggtctcca
      841 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgatgtca
      901 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      961 ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1021 ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1081 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaagacg
     1141 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1201 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1261 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1321 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
     1381 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     1441 accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1501 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc tctggtgcta
     1561 ccaacttctc actgttgaaa caggcagggg atgtagagga gaatccaggg cctggtgcta
     1621 gtggagacta caaagaccat gacggagatt ataaagatca tgacatcgat tacaaggatg
     1681 acgatgacaa gtccggactc agatctggag gctccggagg cccagagcca gcgaagtctg
     1741 ctcccgcccc gaaaaagggc tccaagaagg cggtgactaa ggcgcagaag aaaggcggca
     1801 agaagcgcaa gcgcagccgc aaggagagct attccatcta tgtgtacaag gttctgaagc
     1861 aggtccaccc tgacaccggc atttcgtcca aggccatggg catcatgaat tcgtttgtga
     1921 acgacatttt cgagcgcatc gcaggagagg cttcccgcct ggcgcattac aacaagcgct
     1981 cgaccatcac ctcccgggag atccagacgg ccgtgcgcct gctgctgcct ggggagttgg
     2041 ccaagcacgc cgtgtccgag ggtactaagg ccatcaccaa gtacaccagc gctaagtaat
     2101 gaggcgcgcc gcggccgcga attcgatatc ataatcaacc tctggattac aaaatttgtg
     2161 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     2221 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     2281 aatcctggtt agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     2341 caggggctcg gctgttgggc actgacaatt ccgtggctcg agagatcttc gactgtgcct
     2401 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2461 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2521 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2581 aatagcaggc atgagatctc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     2641 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     2701 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     2761 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     2821 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2881 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     2941 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3001 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     3061 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3121 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     3181 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     3241 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     3301 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     3361 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3421 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     3481 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     3541 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3601 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3661 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3721 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3781 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3841 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3901 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3961 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4021 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4081 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4141 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4201 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4261 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4321 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4381 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4441 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4501 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4561 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4621 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4681 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4741 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4801 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4861 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4921 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4981 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5041 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5101 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5161 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5221 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5281 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5341 tggccttttg ctcacatgt
//
