LOCUS       sequence-446444-        4925 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 4925)
  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 4925)
  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: HCT4_rAAV-eHGT_1140m-minBglobin-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4925
                     /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      937..946
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(943..945,946..948,949..1659)
                     /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(988..1009)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1249..1268)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1596..1617
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1752..1772)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1959..2070
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2188..2328
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2188..2317
                     /label=AAV2 ITR
     rep_origin      2403..2858
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2490..2509)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2700..2721
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2875..2894)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2994..3016
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3054..3072)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3140..3244
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3245..3313,3314..4105)
                     /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(3463..3482)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4276..4864
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4765..4784
                     /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 gcgtgaatgc catcttcaaa ctgtgatttt tttttttttt
      181 gagcaacaag ccagaataaa gtccaagaaa tatctcatcc aattctctct atttttaatg
      241 cagttctatt atagacagga caaggagtcc tcaaaataaa gtgagcagca cagattgacc
      301 atctaccaat ttatagacag gtgctctttc cttatatcag ttctgcctct gaaatctcta
      361 agtaatttgc tagttgtaga cagacaaatg gaggctttct actgctactg ttcttaggca
      421 gggcagactc acccagtaat gttatcaaca acatccccca cagatatatt tattaccacg
      481 attcattttg ctctgcatca gcaggtccat ctggcagtaa aggattgacc aggtgttctg
      541 gggaaagttg gcacacaaat gccttgcata ttgtgacact aacaggagga agagattttt
      601 aaaatgagag gaaattaaat tattcttggt atttgtgagg ttacaagaca tttttagatg
      661 gtaaaaatta tggaacatct ttttgccttg tttgttgctg atttttggct tggtctgaat
      721 tatcagaatg aggaataaat atccaaaggt aattgagtgt gtacgcatct gtgaagtgaa
      781 taaacaaaac agtgtataaa cctcctaaga attttgcagt gaagcaaacc aaaagaccaa
      841 cccgagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc
      901 tgggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtgag caagggcgag
      961 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1021 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1081 ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc
     1141 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     1201 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1261 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1321 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1381 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     1441 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
     1501 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     1561 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1621 gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg
     1681 ccgcgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     1741 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     1801 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt
     1861 cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     1921 ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc
     1981 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2041 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2101 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga
     2161 gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     2221 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     2281 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     2341 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     2401 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2461 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2521 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2581 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     2641 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2701 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     2761 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     2821 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     2881 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     2941 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3001 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     3061 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3121 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3181 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3241 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3301 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3361 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3421 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3481 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3541 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3601 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3661 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3721 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3781 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3841 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3901 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3961 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4021 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4081 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4141 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4201 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4261 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4321 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4381 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4441 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4501 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4561 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4621 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4681 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4741 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4801 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4861 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4921 catgt
//
