LOCUS       sequence-410527-        4567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing SYFP2 in astrocytes AAV Virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4567)
  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 4567)
  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: AiP12084 -
            pAAV-AiE0373m-minBglobin-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4567
                     /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      579..588
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(585..587,588..590,591..1301)
                     /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(630..651)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(891..910)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1238..1259
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1394..1414)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1601..1712
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1830..1970
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1830..1959
                     /label=AAV2 ITR
     rep_origin      2045..2500
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2132..2151)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2342..2363
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2517..2536)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2636..2658
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2696..2714)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2782..2886
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2887..2955,2956..3747)
                     /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(3105..3124)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3918..4506
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4407..4426
                     /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 gcgtcattcc ttcagagaaa tgtttggggt aagggatgga
      181 agtggaanaa aaaaaaaaaa aaaggaaaga aaaagcaaac aaaagccaac cagccttcca
      241 gctgctgaag gagctggggc acaacccttg aagagtgccc ctgccgggaa cagagaggac
      301 tccagctgcc tgcccactgg gccagagcgc tgccttaaaa gaacttttgc ataaaaggaa
      361 aagaggccaa ggctcccctt ttgctgcagg acacagggcc agtgtccaca agagttgagt
      421 gtcctggcca aagccaaagc tagggatcat tgaaaatgcc tgggcagctg gcatggctga
      481 tcagtgagct cgggctgggc ataaaagtca gggcagagcc atctattgct tacatttgct
      541 tctgggatcc agatctttcg aagctagcgc taccggtcgc caccatggtg agcaagggcg
      601 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      661 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      721 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
      781 gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
      841 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
      901 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
      961 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1021 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     1081 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     1141 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     1201 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1261 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagtcgac ggcgcgccgc
     1321 ggccgcgaat tcgatatcat aatcaacctc tggattacaa aatttgtgaa agattgactg
     1381 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     1441 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag
     1501 ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     1561 tgttgggcac tgacaattcc gtggctcgag agatcttcga ctgtgccttc tagttgccag
     1621 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     1681 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     1741 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     1801 gagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc
     1861 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     1921 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     1981 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     2041 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2101 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     2161 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2221 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     2281 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2341 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     2401 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     2461 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     2521 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     2581 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     2641 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     2701 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     2761 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2821 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     2881 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     2941 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3001 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3061 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3121 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3181 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3241 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3301 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3361 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3421 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3481 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     3541 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3601 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3661 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3721 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3781 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3841 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3901 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3961 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4021 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4081 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4141 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4201 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4261 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4321 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4381 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4441 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4501 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4561 cacatgt
//
