LOCUS       sequence-445761-        5338 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 5338)
  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 5338)
  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: HCT77_rAAV-eHGT_1304m-minBglobin-SYFP2-WPRE3-B...
FEATURES             Location/Qualifiers
     source          1..5338
                     /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      1350..1359
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1356..1358,1359..1361,1362..2072)
                     /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(1401..1422)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1662..1681)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2009..2030
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2165..2185)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2372..2483
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2601..2741
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2601..2730
                     /label=AAV2 ITR
     rep_origin      2816..3271
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2903..2922)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3113..3134
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3288..3307)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3407..3429
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3467..3485)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3553..3657
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3658..3726,3727..4518)
                     /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(3876..3895)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4689..5277
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5178..5197
                     /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 gcgtaggtta atcatgttca attaatttga aatacaagcc
      181 acaaaatgtt tatgaatccc caagactggt tgtttgttac cccaacaaag gattgtacga
      241 tctaacgctt gggttctctt gttcccaagc acaaggcagt gccagcatct ctgatggaag
      301 ccagtgaaga aataccaagg tttccatggt ttgtccagtg gagtggtcac aaaaacacaa
      361 gtcaggacac tgacactcac ttatgctagc agtggataaa gggcatcagc caagcaggga
      421 caccctgtcc aagatgctga cgcattcttt tgggtacatt ttgaaaggaa agaagagcag
      481 cttattattg aactaaatgt ttttattgat cgcccttgag ccagtgccca ctgaccccgc
      541 aaaccataaa tggctgtagc acagaatagt catttccagg caactcttgg agatactgaa
      601 ttctcccacc gctctctcca cagtccctcg gatcctctgc ctcctgggtg tgtagaaaca
      661 aggaaaaagg cggaggttgg gtttccccaa ggttctatcc aggagacctg gaggccatgt
      721 gcactcacag gcacatggca cagctgccca gtgtgaggac cagatcagcc cagagctgga
      781 ggacatcccc acccagcgct ggtggcataa agcagacagg gcgcgggcgc gcaaagacaa
      841 atgaatgttt tgcagcattc tccattaaac aataaagcaa gctgtaaagc ttcaggtgct
      901 gtgtttgctg tgcggtctgc gcagcgcggt ggctccggcc ccgggagccg agtgcagtgg
      961 gccggggagc gcctcttcgg ctgatcccag ggaaagataa ggagccgccg agccaccagc
     1021 tctgtctaga ctcgcgacct cacaagtcaa gcgccttgtt tggattagcc ctccagatct
     1081 gccaacctga tcaactcctg aaagagaagg gcccagaaac accccctttt cctcttctgg
     1141 gctacgacga ttgtcgctct tttaagggat caagggccga cttgcccgcg ggagtggtgg
     1201 ggctggttag accagtgtga ggacccccgt cccaccccca attcccagca ctcctcgagc
     1261 tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc ttctgggatc
     1321 cagatctttc gaagctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt
     1381 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     1441 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct
     1501 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcg
     1561 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     1621 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     1681 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     1741 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     1801 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
     1861 gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
     1921 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga
     1981 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     2041 ggatcactct cggcatggac gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa
     2101 ttcgatatca taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2161 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2221 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca
     2281 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2341 ctgacaattc cgtggctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt
     2401 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2461 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2521 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca
     2581 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     2641 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     2701 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     2761 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     2821 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2881 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2941 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3001 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     3061 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3121 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     3181 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3241 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     3301 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3361 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3421 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     3481 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     3541 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3601 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3661 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3721 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3781 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3841 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3901 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3961 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4021 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4081 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4141 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4201 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4261 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4321 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4381 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4441 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4501 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4561 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4621 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4681 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4741 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4801 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4861 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4921 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4981 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5041 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5101 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5161 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5221 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5281 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
