LOCUS       sequence-411062-        4853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0380h_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 4853)
  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 4853)
  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: AiP14253 -
            pAAV-AiE0380h_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4853
                     /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      865..874
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(871..873,874..876,877..1587)
                     /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(916..937)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1177..1196)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1524..1545
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1680..1700)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1887..1998
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2116..2256
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2116..2245
                     /label=AAV2 ITR
     rep_origin      2331..2786
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2418..2437)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2628..2649
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2803..2822)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2922..2944
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2982..3000)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3068..3172
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3173..3241,3242..4033)
                     /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(3391..3410)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4204..4792
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4693..4712
                     /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 gcgtagccca agagtagggg gtggtgctgt gggccactgt
      181 gcagaggcca ggactgtgaa tgaagtgggc actgctcctg ccccctccca ggcctgcctc
      241 tggccaggcc agcctccatc ccggcattgt ctcaggcatc ttgggcaggg ttcccagagc
      301 tggctgaacc aggggcctgg ccaaacagcc ctcccctgcc ctcccattcc tctagcccaa
      361 gagtaggggg tggtgctgtg ggccactgtg cagaggccag gactgtgaat gaagtgggca
      421 ctgctcctgc cccctcccag gcctgcctct ggccaggcca gcctccatcc cggcattgtc
      481 tcaggcatct tgggcagggt tcccagagct ggctgaacca ggggcctggc caaacagccc
      541 tcccctgccc tcccattcct ctagcccaag agtagggggt ggtgctgtgg gccactgtgc
      601 agaggccagg actgtgaatg aagtgggcac tgctcctgcc ccctcccagg cctgcctctg
      661 gccaggccag cctccatccc ggcattgtct caggcatctt gggcagggtt cccagagctg
      721 gctgaaccag gggcctggcc aaacagccct cccctgccct cccattcctc tgagctcggg
      781 ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat
      841 ctttcgaagc tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
      901 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
      961 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
     1021 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag
     1081 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1141 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1201 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1261 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1321 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     1381 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1441 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa
     1501 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1561 actctcggca tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc gcgaattcga
     1621 tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     1681 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     1741 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg
     1801 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     1861 aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat ctgttgtttg
     1921 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1981 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2041 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga tctcacgtgc
     2101 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     2161 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     2221 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     2281 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     2341 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2401 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2461 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2521 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     2581 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2641 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     2701 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2761 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     2821 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     2881 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     2941 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     3001 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3061 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3121 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3181 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3241 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3301 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3361 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3421 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3481 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3541 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3601 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3661 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3721 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3781 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3841 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3901 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3961 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4021 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4081 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4141 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4201 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4261 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4321 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4381 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4441 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4501 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4561 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4621 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4681 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4741 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4801 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
