LOCUS       sequence-325766-        5212 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing SYFP2 AAV Virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5212)
  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 5212)
  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: AiP11525 -
            pAAV-hsA2-eHGT_079h-minRho-SYFP2-WP...
FEATURES             Location/Qualifiers
     source          1..5212
                     /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      1249..1258
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1255..1257,1258..1260,1261..1971)
                     /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(1300..1321)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1561..1580)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1908..1929
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2039..2059)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2246..2357
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2475..2615
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2475..2604
                     /label=AAV2 ITR
     rep_origin      2690..3145
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2777..2796)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2987..3008
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3162..3181)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3281..3303
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3341..3359)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3427..3531
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3532..3600,3601..4392)
                     /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(3750..3769)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4563..5151
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5052..5071
                     /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 gcggccgcaa cgcgtttaga acaatggctg gcccatagta aatgccgtgt
      181 tagtgtgtta gttgctgttc ttccacgtca gaagaggcac agacaaatta ccaccaggtg
      241 gcgctcagag tctgcggagg catcacaaca gccctgaatt tgaatcctgc tctgccactg
      301 cctagttgag accttttact acctgactag ctgtttgtgt attttaggtg tttgtttctg
      361 cagtggggtc aggagtaagt gaggtgaata agatacccag gatatgaaat ttaaagaggc
      421 attcactttc agagtcctgc aaatgcactt acataagtga gtgcctcttt taattttgta
      481 ctctaattga ctcacttcat ctcattctcg tccactgcta gctttgaagg cagagaaaac
      541 gagccatgag ccaaagattg tgggtggcct ccgaaagcta tatatcctga gctgacagcc
      601 agcaaggaaa tgaggacctc agtagaataa ccacatggaa ctgaattcta ccaatagcaa
      661 gagtgatatt ggaagtggat tcattcttag aactcccaga aaggaataca gcctgcctgg
      721 cacttctatt tcaatactgt ggcactctaa gcagaggact ccactgaacc acaccgtacc
      781 cataaacagg aagtctttta tgcagctggt atttatggga attggaaatt ttgacctaca
      841 cagctgtgag ataataaatg ggtgctgttt taagccacta aaattttttt attttttaaa
      901 tttattttta atttccatag gtttttgggg aacaggggta tttggttaca tgagtaagtt
      961 ctttagtggt gattggtgag attttggtgc acctatcacc caagcagtat acactgaacc
     1021 caatttgtag tcttttatcc ctcactcccc tcctacgaat tcgatatcat aatcaaccat
     1081 aggtaccgag ctcgggattc agccgggagc ttagggaggg gaggtcactt cataagggcc
     1141 tgggggggga gttggagcca cgagtcgtcc agccggagcc ccgtgtggct gagctccggc
     1201 ctcagaagca tccccgggtt ggatccttcg aagctagcgc taccggtcgc caccatggtg
     1261 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1321 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1381 ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1441 accaccctgg gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     1501 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1561 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1621 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1681 gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc
     1741 aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac
     1801 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1861 agctaccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1921 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagtcgac
     1981 atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2041 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2101 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg
     2161 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     2221 attccgtggc tcgagagatc ttcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2281 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2341 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2401 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat ctcacgtgcg
     2461 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2521 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     2581 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     2641 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     2701 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2761 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2821 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2881 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     2941 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3001 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     3061 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgggaatttt
     3121 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     3181 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3241 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3301 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3361 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3421 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3481 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3541 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3601 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3661 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3721 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3781 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3841 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3901 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3961 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4021 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4081 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4141 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4201 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4261 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4321 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4381 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4441 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4501 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4561 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4621 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4681 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4741 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4801 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4861 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4921 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4981 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5041 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5101 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5161 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
