LOCUS       sequence-410552-        5410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing SYFP2 in astrocytes AAV Virus. Alias: AiP1347 - 
            pAAV-AiE2118m-minBG-SYFP2-WPRE-BGHpA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5410)
  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 5410)
  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: AiP1347 -
            pAAV-AiE2E118m-minBGpromoter-SYFP2-W...
FEATURES             Location/Qualifiers
     source          1..5410
                     /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      1422..1431
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1428..1430,1431..1433,1434..2144)
                     /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(1473..1494)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1734..1753)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2081..2102
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2237..2257)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2444..2555
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2673..2813
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2673..2802
                     /label=AAV2 ITR
     rep_origin      2888..3343
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2975..2994)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3185..3206
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3360..3379)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3479..3501
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3539..3557)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3625..3729
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3730..3798,3799..4590)
                     /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(3948..3967)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4761..5349
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5250..5269
                     /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 gcgtccaatg tcccaggtga aaatgaacac ttgcaggttt
      181 cttcttccag tcccagccct ctggcccaca ccccactgct gaccacagga gcctctcaca
      241 gtgctcactc agggcttcaa aactctctcc ctggcagctt ctttctattc tgaagtctcc
      301 cagtccagca caaattagtc ccctctcctg gactcctact gcgccgcctt ggtacatcac
      361 agagcactgg ctgagaaaat cgccctttag tccccgcaaa tcactctctt acctgctcca
      421 ctgactaggc ctgtgccatt atttattagg gaatgagcgc tagctgttga gggtcaatta
      481 ttccattgac caggcattac aataattcct ggtaattaaa agaggtactt atgtgaatct
      541 ggatgcgtat tgaaagaaac attagtcctt ttgtcagctt ggcaagtcta ttgttctgag
      601 ccaggcccaa ccaattaaca tcttttgcca atccctgtca gcaggggctt gcaaggagga
      661 gagaaagggg ggccggtcag atgctaattt agatacaatt gtgtcagggt gcatgggagg
      721 ggtaactctg aagaagagac tccagtttaa tgaagcaaac agcttgggtc ccagtgaccc
      781 tgcaggggta attttaaaag gcccctctgt gttcctgtgc caaattgcag agggccagca
      841 acaagaagcc cttcaaaaaa aaaaaaagag ggagcaggat aatcatccac ttaatctgct
      901 cattaaacaa atctctcagg cggacttaac gtgaattagc acttctctga aaggggcagc
      961 cttgatgagc agcagacagc tcagatcatt tggaaggtca cagttttttt tttattatta
     1021 tataagccca gttgttatta atgtattcat atttgaagaa actgagcaga gacctgcttt
     1081 cattcaatct agtgagcaaa acagtacgtt ttcctaactg gaaaacaaaa cataaatgct
     1141 aatcaggtcc cagcagagat caaatcaagc taaatatagc aggcaccctt tgtggttttt
     1201 tttgttttgt tttgttttgt tttttgtttt tttttaatac agaaattaag gaaagaaaaa
     1261 aaaccattga attaattgtc ttcctctgtg cttcgcacct gacaccagcg aggctcacac
     1321 aagggccgga gctcgggctg ggcataaaag tcagggcaga gccatctatt gcttacattt
     1381 gcttctggga tccagatctt tcgaagctag cgctaccggt cgccaccatg gtgagcaagg
     1441 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1501 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1561 tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     1621 tgggctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     1681 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1741 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1801 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1861 actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca
     1921 acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc
     1981 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     2041 agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     2101 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagtc gacggcgcgc
     2161 cgcggccgcg aattcgatat cataatcaac ctctggatta caaaatttgt gaaagattga
     2221 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     2281 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     2341 tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     2401 ggctgttggg cactgacaat tccgtggctc gagagatctt cgactgtgcc ttctagttgc
     2461 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2521 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2581 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2641 catgagatct cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     2701 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     2761 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     2821 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     2881 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2941 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     3001 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3061 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     3121 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3181 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     3241 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     3301 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     3361 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     3421 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3481 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     3541 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     3601 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3661 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3721 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3781 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3841 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3901 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3961 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4021 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4081 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4141 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4201 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4261 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4321 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4381 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4441 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4501 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4561 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4621 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4681 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4741 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4801 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4861 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4921 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4981 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5041 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5101 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5161 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5221 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5281 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5341 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5401 gctcacatgt
//
