LOCUS       sequence-447432-        4864 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 4864)
  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 4864)
  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: HCT15_rAAV-eHGT_1052m-minBglobin-SYFP2-WPRE3-B...
FEATURES             Location/Qualifiers
     source          1..4864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   6..135
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      881..890
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(887..889,890..892,893..1603)
                     /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(932..953)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1193..1212)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1540..1561
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1696..1716)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1903..2014
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2132..2272
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2132..2261
                     /label=AAV2 ITR
     rep_origin      2347..2802
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2434..2453)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2644..2665
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2819..2838)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2938..2960
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2998..3016)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3084..3188
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3189..3257,3258..4049)
                     /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(3407..3426)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4220..4808
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4709..4728
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
       61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      121 tcactagggg ttcctgcggc cgcacgcgta gctctttcac actgtgggtt tttcttttta
      181 tacctcctcc cattcctccc cttctcttct ttcgccttct cctccttttt tatactacag
      241 aaatatgctt ttgctaatga acacatactt tgtagcttct gatttagaat tattaataga
      301 taatctgttt tctgggtctg tcttggacac ttaaaaactg taaatgtcct tgttctattt
      361 gtcatgaaca gcctgtagca aatccctaat caaatgaaag cagcctggga tgtttcataa
      421 gtctacagct aatgtcttta tctctgtagc tttcaagcct ctgacttgga gttcagtgga
      481 agtagccttt ttggacttct tcccttggtg actcagagtc tctggaatca ataaggtctg
      541 cattacttta cagcacaagc taatttaatt catgagaact gtagaagcca aaaggcatct
      601 aatcagagtc actaaaatat taaagcagat aaattataac tctcaggttt acatagactg
      661 tgccatattc catcatcact tccacactgg gttcatgtgc attggagtat gggtcagctc
      721 tgtattcatg cttcttctaa atgtagttca tctcccctcc ccctccccca taggacacag
      781 tgacaaagag ctcgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg
      841 cttctgggat ccagatcttt cgaagctagc gctaccggtc gccaccatgg tgagcaaggg
      901 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      961 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1021 gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     1081 gggctacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     1141 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1201 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1261 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1321 ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     1381 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca
     1441 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
     1501 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1561 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc
     1621 gcggccgcga attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac
     1681 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     1741 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     1801 agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     1861 gctgttgggc actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc
     1921 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     1981 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2041 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2101 atgagatctc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     2161 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     2221 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     2281 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     2341 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2401 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     2461 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2521 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     2581 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2641 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     2701 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     2761 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     2821 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     2881 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     2941 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     3001 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     3061 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3121 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3181 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3241 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3301 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3361 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3421 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3481 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3541 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3601 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3661 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3721 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3781 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3841 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3901 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3961 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4021 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4081 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4141 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4201 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4261 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4321 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4381 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4441 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4501 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4561 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4621 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4681 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4741 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4801 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4861 ctca
//
