LOCUS       sequence-410469-        5173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0361h is an enhancer sequence designed to drive AAV-mediated 
            transgene expression in brain astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5173)
  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 5173)
  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: AiP12979 - pAAV-AiE0361h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..5173
                     /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      1185..1194
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1191..1193,1194..1196,1197..1907)
                     /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(1236..1257)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1497..1516)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1844..1865
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2000..2020)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2207..2318
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2436..2576
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2436..2565
                     /label=AAV2 ITR
     rep_origin      2651..3106
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2738..2757)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2948..2969
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3123..3142)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3242..3264
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3302..3320)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3388..3492
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3493..3561,3562..4353)
                     /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(3711..3730)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4524..5112
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5013..5032
                     /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 gcgtgtccag atgctgttgc cgtggtaata ctaaatcgtg
      181 tccaaagagg aggctgagat tcctttttag ttaaccttgt ccacatgatt tgactatcca
      241 gcaggcttgg attcccaggc cccagagact tcgggactgt tttccagcac ttgaatgtgg
      301 tatataagtg ctgatatcaa aagggatatg taggaaagac aagattatag gcagggcatt
      361 gtctgaccag gacgaggccc gaagcaaaag tctttgtttg tctctggcac ctcttgataa
      421 ttgttgtttt actcctagaa cccacccact gatccccatc catccctagt accagtcacg
      481 gctgctctag catttcggcc ttgtcaacta ctgcaggccc tgatcaagga ggcctctgtt
      541 catggggtaa ggttaagggg tgggttctgg gaggagggta agactgtagt gcagattcga
      601 gacctaggga ggtggggcca gtttctcact caatgagcct gcctgttgtt tagctctatt
      661 gtatagggtt cttctatctc tccctttgtc agctggattc aagggacagg ctgggaattg
      721 tgcagggtac aattggtctc tttctaccat tcctttcgat gaaagccctt cctctccttc
      781 ctttgatgag ttcccagatt cacactgaat ttccatcatg gagctggtcc ttgaggcggg
      841 cccagggcat gggaaaggag ggagggaggg ttgggagctc ttggtgcctg cttaaagaga
      901 cagcatcagg atcaacacag tcccttgatg aacctggatc tgtgtaagtc acattggttc
      961 tgttgatctc tgtcagaccg ctgtgaagag cgctggtgag aagggcttaa agttggagta
     1021 tcaactatca cagattgatt agacttgttt tgggagtttg tatacaaatg tagccaacca
     1081 tgacatgatc tgagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca
     1141 tttgcttctg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc atggtgagca
     1201 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1261 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1321 ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1381 ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1441 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1501 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1561 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1621 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1681 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
     1741 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     1801 accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1861 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa gtcgacggcg
     1921 cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat
     1981 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2041 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2101 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     2161 ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt
     2221 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2281 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2341 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2401 aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     2461 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     2521 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     2581 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     2641 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2701 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     2761 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2821 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     2881 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2941 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     3001 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     3061 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     3121 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     3181 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     3241 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     3301 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     3361 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3421 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3481 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3541 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3601 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3661 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3721 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3781 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3841 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3901 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3961 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4021 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4081 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4141 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4201 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4261 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4321 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4381 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4441 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4501 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4561 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4621 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4681 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4741 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4801 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4861 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4921 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4981 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5041 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5101 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5161 tttgctcaca tgt
//
