LOCUS       sequence-411036-        5394 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 5394)
  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 5394)
  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: AiP13972 -
            pAAV-GfaABC1D-SYFP2-P2A-3XFLAG-10aa...
FEATURES             Location/Qualifiers
     source          1..5394
                     /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      860..869
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(866..868,869..871,872..1582)
                     /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(1172..1191)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1519..1540
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1592..1648
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1661..1726
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1757..2134
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2221..2241)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2428..2539
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2657..2797
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2657..2786
                     /label=AAV2 ITR
     rep_origin      2872..3327
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2959..2978)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3169..3190
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3344..3363)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3463..3485
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3523..3541)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3609..3713
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3714..3782,3783..4574)
                     /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"
     rep_origin      4745..5333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5234..5253
                     /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 gcgtaacata tcctggtgtg gagtagggga cgctgctctg
      181 acagaggctc gggggcctga gctggctctg tgagctgggg aggaggcaga cagccaggcc
      241 ttgtctgcaa gcagacctgg cagcattggg ctggccgccc cccagggcct cctcttcatg
      301 cccagtgaat gactcacctt ggcacagaca caatgttcgg ggtgggcaca gtgcctgctt
      361 cccgccgcac cccagccccc ctcaaatgcc ttccgagaag cccattgagc agggggcttg
      421 cattgcaccc cagcctgaca gcctggcatc ttgggataaa agcagcacag ccccctaggg
      481 gctgcccttg ctgtgtggcg ccaccggcgg tggagaacaa ggctctattc agcctgtgcc
      541 caggaaaggg gatcagggga tgcccaggca tggacagtgg gtggcagggg gggagaggag
      601 ggctgtctgc ttcccagaag tccaaggaca caaatgggtg aggggagagc tctccccata
      661 gctgggctgc ggcccaaccc caccccctca ggctatgcca gggggtgttg ccaggggcac
      721 ccgggcatcg ccagtctagc ccactccttc ataaagccct cgcatcccag gagcgagcag
      781 agccagagca ggttggagag gagacgcatc acctccgctg ctcgcggatc cagatctttc
      841 gaagctagcg ctaccggtcg ccaccatggt ctccaagggc gaggagctgt tcaccggggt
      901 ggtgcccatc ctggtcgagc tggacggcga tgtcaacggc cacaagttca gcgtgtccgg
      961 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg
     1021 caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt
     1081 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1141 ctacgtccag gagcgcacca tcttcttcaa agacgacggc aactacaaga cccgcgccga
     1201 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1261 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     1321 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
     1381 cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1441 ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc
     1501 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1561 cggcatggac gagctgtaca agggctctgg tgctaccaac ttctcactgt tgaaacaggc
     1621 aggggatgta gaggagaatc cagggcctgg tgctagtgga gactacaaag accatgacgg
     1681 agattataaa gatcatgaca tcgattacaa ggatgacgat gacaagtccg gactcagatc
     1741 tggaggctcc ggaggcccag agccagcgaa gtctgctccc gccccgaaaa agggctccaa
     1801 gaaggcggtg actaaggcgc agaagaaagg cggcaagaag cgcaagcgca gccgcaagga
     1861 gagctattcc atctatgtgt ataaggttct gaagcaggtc caccctgaca ccggcatttc
     1921 gtccaaggcc atgggcatca tgaactcgtt tgtgaacgac attttcgagc gcatcgcagg
     1981 tgaggcttcc cgcctggcgc attacaacaa gcgctcgacc atcacctcca gggagatcca
     2041 gacggccgtg cgcctgctgc tgcctgggga gttggccaag cacgccgtgt ccgagggtac
     2101 taaggccatc accaagtaca ccagcgctaa gtaatgaggc gcgccgcggc cgcgaattcg
     2161 atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2221 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2281 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc
     2341 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2401 caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt
     2461 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2521 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2581 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg
     2641 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     2701 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     2761 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     2821 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     2881 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2941 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3001 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3061 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     3121 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3181 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     3241 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3301 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     3361 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3421 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3481 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3541 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4861 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
