LOCUS       sequence-410589-        4961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0390h_3xC2 is an optimized 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 4961)
  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 4961)
  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: AiP13572 -
            pAAV-AiE0390h_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4961
                     /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      973..982
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(979..981,982..984,985..1695)
                     /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(1024..1045)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1285..1304)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1632..1653
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1788..1808)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1995..2106
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2224..2364
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2224..2353
                     /label=AAV2 ITR
     rep_origin      2439..2894
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2526..2545)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2736..2757
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2911..2930)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3030..3052
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3090..3108)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3176..3280
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3281..3349,3350..4141)
                     /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(3499..3518)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4312..4900
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4801..4820
                     /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 gcgtctagcc gctcagctca attcacccca cacaaaggct
      181 ggagcctaga cctcaatgga ccgagtgaaa catgttcaaa actaggctct ctattgtgac
      241 tgaatttctt aacatctttt caaaaagcgg agaatgcctt gaggctaaag gaagaaacag
      301 gctaatggtg aattgggaat tctgagcaaa tttcagagcc ctttcctcct agcttttgag
      361 gttgaaagca agctctttcc tttcaagttc tagccgctca gctcaattca ccccacacaa
      421 aggctggagc ctagacctca atggaccgag tgaaacatgt tcaaaactag gctctctatt
      481 gtgactgaat ttcttaacat cttttcaaaa agcggagaat gccttgaggc taaaggaaga
      541 aacaggctaa tggtgaattg ggaattctga gcaaatttca gagccctttc ctcctagctt
      601 ttgaggttga aagcaagctc tttcctttca agttctagcc gctcagctca attcacccca
      661 cacaaaggct ggagcctaga cctcaatgga ccgagtgaaa catgttcaaa actaggctct
      721 ctattgtgac tgaatttctt aacatctttt caaaaagcgg agaatgcctt gaggctaaag
      781 gaagaaacag gctaatggtg aattgggaat tctgagcaaa tttcagagcc ctttcctcct
      841 agcttttgag gttgaaagca agctctttcc tttcaagttg agctcgggct gggcataaaa
      901 gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct ttcgaagcta
      961 gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     1021 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     1081 gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg
     1141 cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc
     1201 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     1261 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     1321 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1381 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc
     1441 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac
     1501 ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1561 ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag
     1621 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1681 gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc gaattcgata tcataatcaa
     1741 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     1801 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     1861 ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc
     1921 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct
     1981 cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2041 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2101 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     2161 caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc
     2221 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     2281 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     2341 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     2401 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     2461 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     2521 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2581 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2641 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2701 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2761 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     2821 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     2881 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2941 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3001 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3061 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     3121 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     3181 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3241 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3301 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3361 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3421 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3481 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3541 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3601 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3661 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3721 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3781 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3841 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3901 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3961 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4021 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4081 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4141 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4201 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4261 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4321 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4381 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4441 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4501 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4561 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4621 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4681 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4741 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4801 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4861 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4921 tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
