LOCUS       sequence-444325-        5374 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 5374)
  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 5374)
  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: AiP1351 -
            pAAV-AiE2E122m-minBGpromoter-SYFP2-W...
FEATURES             Location/Qualifiers
     source          1..5374
                     /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      1386..1395
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1392..1394,1395..1397,1398..2108)
                     /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(1437..1458)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1698..1717)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2045..2066
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2201..2221)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2408..2519
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2637..2777
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2637..2766
                     /label=AAV2 ITR
     rep_origin      2852..3307
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2939..2958)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3149..3170
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3324..3343)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3443..3465
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3503..3521)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3589..3693
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3694..3762,3763..4554)
                     /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(3912..3931)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4725..5313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5214..5233
                     /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 gcgttaaggc tatagtcctt ggtgttgaaa gactcattca
      181 tttctctctt atcttcaatg ttagggactg attcaaaaca gcataatcaa aacaaagcag
      241 agcacagcat aattaaaatc atgtcttctg aataggtaat tacaaaacta attggatatt
      301 ggtaattatt ttctttttac ctgttcctgg ctcctttctc agttaatatg tagtgctatg
      361 tacagatgaa gttcagatgc aattatgtac agggtctatc tcaccaccag gaagcaggac
      421 ttttctgtgt gatcttccat ttcctgcaag gactacagaa ttgaggtaca agtgcctttt
      481 gttttcctca agttgagtcc ttctgttttc tttgtcactg gctttcatcc tctgtcccac
      541 tggctgtggc tcagtgttta tttattggct gtttagtatg gagaaacatc cagtctttca
      601 acattgacaa caaatccaaa taatgagcct gcccatcacg ctacactggg tatcatcttt
      661 gcctgttagg ctcttggctg gttttttgct gtatttgact atcctcaaag tttgggagtt
      721 ctgactgcgg gttgttattt tccaattctt attaacactt gccaaatctt ttgtcatgca
      781 tctttatact catgacagct tctaggtcaa attttagttt tgatcactgg atctttagaa
      841 ggaaagctaa ggaactgaca gacatagcca gctccttttt ttatgctggc aatggatact
      901 gattagctgt gtgctagttg ttgaagtcat agcaacaagt cctacttact atcttttagg
      961 catctcaatt aagtttcttt ggtctttgga aatataacta caattaagtt tcatctcctg
     1021 gagaaaatga gtatgttcag gagaacaact tcatagagtt aattaagaac tcttttctag
     1081 agttcatttt taaactggta agttatttgt actccttcat tttccttttg tttccaggtt
     1141 ttataatagg cttttgatgg atttaatctt ttaatttcaa actgactatt tgccaagatt
     1201 ctggatgcct ttgctgaggc taccaagtta agtatatgca tcagcaacca gaggaaaaac
     1261 actaacgttg ggcagagaag caatgaatac cggagctcgg gctgggcata aaagtcaggg
     1321 cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac
     1381 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1441 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1501 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
     1561 cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg
     1621 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1681 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1741 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1801 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
     1861 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
     1921 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1981 ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg
     2041 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2101 tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg atatcataat caacctctgg
     2161 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     2221 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     2281 tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc gccgcctgcc
     2341 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gctcgagaga
     2401 tcttcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2461 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2521 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2581 gaggattggg aagacaatag caggcatgag atctcacgtg cggaccgagc ggccgcagga
     2641 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     2701 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     2761 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2821 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     2881 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     2941 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3001 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3061 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3121 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3181 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     3241 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     3301 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     3361 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3421 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     3481 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     3541 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3601 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3661 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     3721 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     3781 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     3841 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     3901 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     3961 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4021 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4081 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4141 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4201 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     4261 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     4321 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     4381 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     4441 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     4501 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     4561 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4621 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4681 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4741 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4801 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4861 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4921 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4981 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5041 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5101 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5161 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5221 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5281 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5341 ttcctggcct tttgctggcc ttttgctcac atgt
//
