LOCUS       sequence-325759-        4700 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 4700)
  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 4700)
  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: AiP11253 - pAAV-eHGT_017h-minBG-SYFP2-WPRE3-BG...
FEATURES             Location/Qualifiers
     source          1..4700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(296..312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(296..312)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(309..331)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    320..336
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(344..350,351..368,369..374))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    389..410
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(698..1286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(778..797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1457..2248,2249..2317))
                     /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     2080..2099
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2318..2422)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2490..2508
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2546..2568)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2668..2687
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2881..2903
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2895..2912
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2896..2912
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3151..3262
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3442..3462
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3555..4265,4266..4268,4269..4271))
                     /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(3597..3618)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3946..3965
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4205..4226
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4268..4277
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gtgccgcccc gcccagggtc cctgccaccc tgcacctcca tgctgcccac cccttggccc
       61 tgtgggtgtg acctgcagcc ctcaggctgt taccacgcgt cctaggtcac gactccaccc
      121 ctccaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
      181 gaggccgccc gggcaaagcc cgggcggcct cagtgagcga gcgagcgcgc agagagggag
      241 tggcccagat ccccccggga agctttctag atctcgagag cttggcgtaa tcatggtcat
      301 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
      361 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
      421 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
      481 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
      541 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
      601 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
      661 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
      721 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
      781 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
      841 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
      901 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
      961 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1021 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1081 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     1141 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1201 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1261 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1321 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     1381 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1441 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1501 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     1561 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     1621 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     1681 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     1741 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     1801 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     1861 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     1921 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     1981 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     2041 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     2101 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     2161 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     2221 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     2281 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     2341 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     2401 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     2461 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     2521 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     2581 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     2641 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     2701 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     2761 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     2821 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     2881 cccagtcacg acgttgtaaa acgacggcca gtgaattgtc gacgcatgcg ggcccggatc
     2941 tgtccctctc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg
     3001 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggactag
     3061 tcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
     3121 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
     3181 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
     3241 ggctggcaac tagaaggcac agtcgctcga gccacggaat tgtcagtgcc caacagccga
     3301 gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc aagaactaac
     3361 caggatttat acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa
     3421 ggcattaaag cagcgtatcc acatagcgta aaaggagcaa catagttaag aataccagtc
     3481 aatctttcac aaattttgta atccagaggt tgattatgat atcgaattcg cggccgcggc
     3541 gcgccgtcga cttacttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg
     3601 aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag cttggactgg
     3661 tagctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc
     3721 tggtagtggt cggcgagctg cacgccgccg tcctcgatgt tgtggcggat cttgaagttg
     3781 gccttgatgc cgttcttctg cttgtcggcg gtgatataga cgttgtggct gttgtagttg
     3841 tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg
     3901 atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg
     3961 tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag
     4021 aagtcgtgct gcttcatgtg gtcggggtag cgggcgaagc actgcacgcc gtagcccagg
     4081 gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat cagcttcagg
     4141 gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg
     4201 tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc
     4261 ttgctcacca tggtggcgac cggtagcgct agcttcgaaa gatctggatc ccagaagcaa
     4321 atgtaagcaa tagatggctc tgccctgact tttatgccca gcccgagctc actgtgtgcc
     4381 gtcggaaggg gggcccagca gtgccctcgg gcaggcagtg gcggctgcca cgctggactc
     4441 actcccctgg caccctggcg gttgggggga gctcctcctc ctccttccag gatgaagaac
     4501 aactcatcaa cttgcaaatg tccgggtgag agggttggaa aaatgaaggc gcccaaactt
     4561 gacatttgca tttcaaacac ttatatcagc aaaacaaata acagtaatta taccatttgc
     4621 atattttgtg caatgattgt ttagcgaggt ggggaggagg ggcgcctgct gatgccagca
     4681 gcccagcctc ccgcacagct
//
