LOCUS       sequence-466770-        5295 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 5295)
  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 5295)
  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: 
            HCT47_pAAV-eHGT_3Xcore_1137m-minBGprom-iCre297T-WPRE3-bGHpA
FEATURES             Location/Qualifiers
     source          1..5295
                     /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      1307..1316
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1313..1315,1316..1318,1319..2029)
                     /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(1358..1379)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1619..1638)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1966..1987
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2122..2142)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2329..2440
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2558..2698
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2558..2687
                     /label=AAV2 ITR
     rep_origin      2773..3228
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2860..2879)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3070..3091
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3245..3264)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3364..3386
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3424..3442)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3510..3614
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3615..3683,3684..4475)
                     /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(3833..3852)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4646..5234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5135..5154
                     /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 gcgtgatttc tttgtctctg tggaagcact taagtgcagg
      181 ctttagttcc aatgacactc aggagcctct ggattccagc actggggatg ggggtggggt
      241 agaacgttct caggcctcac caacccctcc cctgtgtgct gcctttggga gagtcccaag
      301 gcttcagcat tacttaatta attaggcctc tactgctaca taggctcaga ttcaaaagaa
      361 cagagtggcc cacgtcagcc attcccggaa aagtctgatg gctggaagcc agaggactat
      421 gtgtctgcct tgctgccctt ggccagccca tcctgaatgc ccagactcgg acaatggagt
      481 aggtacagaa gggtagattt ctttgtctct gtggaagcac ttaagtgcag gctttagttc
      541 caatgacact caggagcctc tggattccag cactggggat gggggtgggg tagaacgttc
      601 tcaggcctca ccaacccctc ccctgtgtgc tgcctttggg agagtcccaa ggcttcagca
      661 ttacttaatt aattaggcct ctactgctac ataggctcag attcaaaaga acagagtggc
      721 ccacgtcagc cattcccgga aaagtctgat ggctggaagc cagaggacta tgtgtctgcc
      781 ttgctgccct tggccagccc atcctgaatg cccagactcg gacaatggag taggtacaga
      841 agggtagatt tctttgtctc tgtggaagca cttaagtgca ggctttagtt ccaatgacac
      901 tcaggagcct ctggattcca gcactgggga tgggggtggg gtagaacgtt ctcaggcctc
      961 accaacccct cccctgtgtg ctgcctttgg gagagtccca aggcttcagc attacttaat
     1021 taattaggcc tctactgcta cataggctca gattcaaaag aacagagtgg cccacgtcag
     1081 ccattcccgg aaaagtctga tggctggaag ccagaggact atgtgtctgc cttgctgccc
     1141 ttggccagcc catcctgaat gcccagactc ggacaatgga gtaggtacag aagggtacta
     1201 tttcactgag tttgagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta
     1261 catttgcttc tgggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtgag
     1321 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1381 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1441 gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1501 caccctgggc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     1561 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1621 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1681 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1741 gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     1801 ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta
     1861 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1921 ctaccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1981 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacgg
     2041 cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag
     2101 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2161 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2221 ctggttagtt cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2281 ggctcggctg ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta
     2341 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2401 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2461 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2521 gcaggcatga gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     2581 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     2641 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     2701 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     2761 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2821 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     2881 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2941 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     3001 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3061 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     3121 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     3181 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     3241 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     3301 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3361 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     3421 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3481 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3541 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3601 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3661 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3721 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3781 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3841 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3901 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3961 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4021 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4081 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4141 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4201 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4261 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4321 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4381 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4441 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4501 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4561 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4621 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4681 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4741 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4801 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4861 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4921 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4981 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5041 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5101 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5161 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5221 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5281 cttttgctca catgt
//
