LOCUS       sequence-410550-        5442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing SYFP2 in astrocytes AAV Virus. Alias: AiP1346 - 
            pAAV-AiE2117m-minBG-SYFP2-WPRE-BGHpA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5442)
  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 5442)
  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: AiP1346 -
            pAAV-AiE2E117m-minBGpromoter-SYFP2-W...
FEATURES             Location/Qualifiers
     source          1..5442
                     /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      1454..1463
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1460..1462,1463..1465,1466..2176)
                     /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(1505..1526)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1766..1785)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2113..2134
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2269..2289)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2476..2587
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2705..2845
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2705..2834
                     /label=AAV2 ITR
     rep_origin      2920..3375
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3007..3026)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3217..3238
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3392..3411)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3511..3533
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3571..3589)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3657..3761
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3762..3830,3831..4622)
                     /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(3980..3999)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4793..5381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5282..5301
                     /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 gcgttctaca agtgtctctt cgaaccaaag gacatgcctg
      181 ctaaagcata ttttataatc tcttaagagc tggagtaagc cttagtgtgg caggtctgcg
      241 tgtcaaataa agggtttgag atgacagaac agcagcgtcc tctcagaagg tccagctgag
      301 cccccagggt gagggggtga catgctgtgc acagtggccc tcatttgagc aggaatgccg
      361 ttcaggtgcc tgctgtgcct ccctggtgcc aggcccagct ggctctctac cattccagcc
      421 ctcctttgcc acatcagttt gttcttaatt ccttctcgga aactctgctc ctttgccgca
      481 caccacgtgt tcagccaggc cccatctgac agctggggcc cctggcgcaa ccccacagac
      541 acccagtgtc cctttcacag gctccggtga gccaagtcta cttcctttac aaccctttta
      601 taggccctcc cattctggag ccttgtgtgc catgccaatg gcttacacac ttgatgaaag
      661 gactcagggg ctgagaactt ggatcaatga ggtctttatt tttgaacatc agtaaacagc
      721 acaaatcagt tgaatgggtg cagccctgtt cgtaattcca atgcttgctg tggtttccag
      781 ccagcattag acacacagag aaagtgctcg ttaatgctag ctaatgaaga atgtggccct
      841 ttttccctaa cttgaacaca ctccaagacg ctggcctgaa agatcagtgt gacactggag
      901 agggtgaatg ggaaaatctt taaccttcct tttgaaacag cccattgacg tggctctaac
      961 cactttcttt cctcttcttt ttggtctgct ctctggcatt gcgggatcag cttgacgcaa
     1021 aatacaatct tttttaagac ccagaaaatc gtgccaggac cgtttcacta actgaactac
     1081 aaatcgtgta attagagatg aaaagtccca ggaaccttca acttgtgcct gtgcctttgt
     1141 caaagtggac ccctgtcact gtgttttggg ggttgttctt gtcactcttg ctgtttattt
     1201 gtgaaaccac tcacagctca ttcagtgcca ccagcaaggg acatggcctc cttgtttcaa
     1261 ctaaaaatag tcattggtgt ggttaaaacc acaggaaagt aatctttaaa aagatttaag
     1321 atctggagtt gtagctcaag ttgtagctag ctttgcctag gagctcgggc tgggcataaa
     1381 agtcagggca gagccatcta ttgcttacat ttgcttctgg gatccagatc tttcgaagct
     1441 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1501 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1561 cgagggcgat gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct
     1621 gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggcgtgcagt gcttcgcccg
     1681 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1741 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1801 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1861 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcac
     1921 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
     1981 cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2041 gctgctgccc gacaaccact acctgagcta ccagtccaag ctgagcaaag accccaacga
     2101 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2161 ggacgagctg tacaagtaag tcgacggcgc gccgcggccg cgaattcgat atcataatca
     2221 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2281 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2341 tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc
     2401 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggc
     2461 tcgagagatc ttcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2521 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2581 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2641 gcaaggggga ggattgggaa gacaatagca ggcatgagat ctcacgtgcg gaccgagcgg
     2701 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     2761 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     2821 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     2881 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     2941 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     3001 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3061 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3121 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3181 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3241 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     3301 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3361 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3421 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3481 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3541 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     3601 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     3661 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3721 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3781 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3841 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3901 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3961 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4021 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4081 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4141 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4201 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4261 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4321 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4381 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4441 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4501 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4561 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4621 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4681 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4741 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4801 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4861 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4921 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4981 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5041 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5101 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5161 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5221 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5281 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5341 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5401 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
