LOCUS       sequence-447420-        5455 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 5455)
  AUTHORS   Kussick E, Johansen N, Taskin N, Chowdhury A, Quinlan MA, Fraser A, 
            Clark AG, Wynalda B, Martinez R, Groce EL, Reding M, Liang E, Shulga
            L, Huang C, Casper T, Clark M, Ho W, Gao Y, van Velthoven CTJ, 
            Sobieski C, Ferrer R, Berg MR, Curtis BC, English C, Day JC, Fortuna
            MG, Donadio N, Newman D, Yao S, Chakka AB, Goldy J, Torkelson A, 
            Guzman JB, Chakrabarty R, Nguy B, Guilford N, Pham TH, Wright V, 
            Ronellenfitch K, Naidoo R, Kenney J, Williford A, Ramakrishnan C, 
            Drinnenberg A, Gudsnuk K, Thyagarajan B, Smith KA, Dee N, Deisseroth
            K, Zeng H, Yao Z, Tasic B, Levi BP, Hodge R, Bakken TE, Lein ES, 
            Ting JT, Daigle TL
  TITLE     Enhancer AAVs for targeting spinal motor neurons and descending 
            motor pathways in rodents and macaque.
  JOURNAL   Cell Rep. 2025 Jun 24;44(6):115730. doi: 
            10.1016/j.celrep.2025.115730. Epub 2025 May 21.
  PUBMED    40403722
REFERENCE   2  (bases 1 to 5455)
  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 Rep.";
            date: "2025-06-24"; volume: "44(6):115730. doi"; pages: " 
            10.1016/j.celrep.2025.115730. Epub 2025 May 21"
COMMENT     Sequence Label: 
            HCT49_rAAV-3Xcore-eHGT_1139m-minBglobin-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5455
                     /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      1467..1476
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1473..1475,1476..1478,1479..2189)
                     /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(1518..1539)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1779..1798)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2126..2147
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2282..2302)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2489..2600
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2718..2858
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2718..2847
                     /label=AAV2 ITR
     rep_origin      2933..3388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3020..3039)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3230..3251
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3405..3424)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3524..3546
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3584..3602)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3670..3774
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3775..3843,3844..4635)
                     /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(3993..4012)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4806..5394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5295..5314
                     /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 gcgtaaatat tacaatacta aaaaaggaaa gacaagaaat
      181 gaagccagta gatatatttt caatagcata gtatatttcc agttaccaca tttttttgct
      241 gatgtttgta cttaaatgta tattaaaaag tttcttatcc caagtaaaga gcattactaa
      301 aattactgtg tgccactctg aaagatgaga gaaacccata ttcagtttcc ataatgaaat
      361 ccatatttgc ttggttgtga tccaaattgt ttttgaaaag tatatattta attgaggcag
      421 atgaagtggt ccaaaattta ataagtatta aaaaaaagga aaatttaaag agaaaatttc
      481 caattaaaga tgagtacctg atgattgctc aggcccaatt attaatattg attaatgttt
      541 tgtactttaa atattacaat actaaaaaag gaaagacaag aaatgaagcc agtagatata
      601 ttttcaatag catagtatat ttccagttac cacatttttt tgctgatgtt tgtacttaaa
      661 tgtatattaa aaagtttctt atcccaagta aagagcatta ctaaaattac tgtgtgccac
      721 tctgaaagat gagagaaacc catattcagt ttccataatg aaatccatat ttgcttggtt
      781 gtgatccaaa ttgtttttga aaagtatata tttaattgag gcagatgaag tggtccaaaa
      841 tttaataagt attaaaaaaa aggaaaattt aaagagaaaa tttccaatta aagatgagta
      901 cctgatgatt gctcaggccc aattattaat attgattaat gttttgtact ttaaatatta
      961 caatactaaa aaaggaaaga caagaaatga agccagtaga tatattttca atagcatagt
     1021 atatttccag ttaccacatt tttttgctga tgtttgtact taaatgtata ttaaaaagtt
     1081 tcttatccca agtaaagagc attactaaaa ttactgtgtg ccactctgaa agatgagaga
     1141 aacccatatt cagtttccat aatgaaatcc atatttgctt ggttgtgatc caaattgttt
     1201 ttgaaaagta tatatttaat tgaggcagat gaagtggtcc aaaatttaat aagtattaaa
     1261 aaaaaggaaa atttaaagag aaaatttcca attaaagatg agtacctgat gattgctcag
     1321 gcccaattat taatattgat taatgttttg tactttacta tttcactgag tttgagctcg
     1381 ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc tgggatccag
     1441 atctttcgaa gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca
     1501 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1561 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ctgatctgca
     1621 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc tacggcgtgc
     1681 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1741 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1801 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1861 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1921 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     1981 acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     2041 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc aagctgagca
     2101 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     2161 tcactctcgg catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg ccgcgaattc
     2221 gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2281 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2341 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt cttgccacgg
     2401 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     2461 acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc atctgttgtt
     2521 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2581 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2641 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga gatctcacgt
     2701 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     2761 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     2821 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     2881 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     2941 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3001 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3061 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3121 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     3181 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3241 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     3301 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3361 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     3421 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3481 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3541 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     3601 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3661 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3721 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3781 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3841 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3901 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3961 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4021 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4081 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4141 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4201 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4261 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4321 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4381 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4441 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4501 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4561 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4621 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4681 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4741 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4801 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4861 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4921 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4981 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5041 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5101 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5161 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5221 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5281 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5341 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5401 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
