LOCUS       sequence-325817-        4769 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 4769)
  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 4769)
  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: AiP11244 - pAAV-hDLXI56i-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(228..247)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     575..596
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(731..751)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    938..1049
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1167..1307
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1167..1296
                     /label=AAV2 ITR
     rep_origin      1382..1837
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1469..1488)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1679..1700
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1854..1873)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1973..1995
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2033..2051)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2119..2223
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2224..2292,2293..3084)
                     /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(2442..2461)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3255..3843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3744..3763
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3905..4034
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      4685..4694
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4691..4693,4694..4696,4697..4769,1..638)
                     /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(4736..4757)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc
       61 tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct
      121 acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
      181 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
      241 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
      301 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
      361 acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
      421 agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca
      481 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca
      541 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
      601 ccgccgggat cactctcggc atggacgagc tgtacaagta agtcgacggc gcgccgcggc
      661 cgcgaattcg atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
      721 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
      781 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc
      841 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
      901 tgggcactga caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca
      961 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     1021 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     1081 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag
     1141 atctcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
     1201 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     1261 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     1321 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     1381 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     1441 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     1501 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     1561 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     1621 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     1681 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     1741 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     1801 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     1861 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     1921 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     1981 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     2041 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     2101 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2161 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2221 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2281 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2341 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2401 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2461 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2521 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2581 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2641 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2701 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2761 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2821 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2881 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2941 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3001 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3061 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3121 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3181 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3241 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3301 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3361 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     3421 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3481 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3541 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3601 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3661 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3721 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3781 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3841 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3901 atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac
     3961 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
     4021 cactaggggt tcctgcggcc gcacgcgtat aggtaccgag ctctatgcac tcacagtggt
     4081 ttggcatgca tctggtgaat tttttttaac gaaaaattag tgttggtttc gatgtatggt
     4141 agcattctcc ctaacgtaat ttgaataatt cagcaaagcc ccactaccag ctgtacttct
     4201 gcagcctctt ccattctttt cagcattata attttggtta attttcaatt ttaggtccta
     4261 cgtctctgca atttgtgtat gaataacaga ataatttccc tcttttgttt cgcctttcct
     4321 gttcctgaat ctaaataaag atggcttttt agtattaaaa gtggaagaaa attacaggta
     4381 attatctttg acggtaaaaa cgctgtaatc agcgggctac atgaaaaatt actctaatta
     4441 tggctgcatt taagagaatg gaaaaaaacc ttcttgtgga taaaaacctt aaattgtccc
     4501 caatgtctgc ttcaaattgg atggcactgc agctggaggc tttgttcaga attgatcctg
     4561 gggagctacg aacccaaagt ttcacagtag ggagctcggg ctgggcataa aagtcagggc
     4621 agagccatct attgcttaca tttgcttctg ggatccagat ctttcgaagc tagcgctacc
     4681 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     4741 cgagctggac ggcgacgtaa acggccaca
//
