LOCUS       sequence-447423-        5930 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 5930)
  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 5930)
  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: HCT10_rAAV-eHGT_1145m-minBglobin-SYFP2-WPRE3-B...
FEATURES             Location/Qualifiers
     source          1..5930
                     /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      1942..1951
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1948..1950,1951..1953,1954..2664)
                     /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(1993..2014)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2254..2273)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2601..2622
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2757..2777)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2964..3075
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3193..3333
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3193..3322
                     /label=AAV2 ITR
     rep_origin      3408..3863
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3495..3514)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3705..3726
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3880..3899)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3999..4021
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4059..4077)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4145..4249
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4250..4318,4319..5110)
                     /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(4468..4487)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5281..5869
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5770..5789
                     /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 gcgttgtaag agtaataatc gccggcgctg ccaagagccc
      181 aatgcctccc tgatccattc cctcaatgaa tacgagtgtg tttggagagg gggttagact
      241 gggaggggtc acggagaagg gattccagca aaggaattca gggcggggga cgggctaggg
      301 tactcattag aggaggtggg ccccaagcca aagttggcca gcgctcttga agaatgacca
      361 tcttgaccta aatggggagg gtaggtggcc tggaggagaa gcgggtctgc tagctgcggg
      421 agtagtagtt gtagtcgtcc tcacagccat caaaagggcc aggcgggcta caaggttcac
      481 cgccgtcctt gccctgcacc tcacgcaggc gcatcgcgtc gtacagaccc tgcggcggtt
      541 catcaagcag cacatcgcgc tcggaacgcg agcgtgtaag gaggcccaca ttacgcaaga
      601 gcaataggac cggtgcgtag agcaggttgg ccaggcccat gccaaggctg agctgctcaa
      661 agccaagtga gtgaacgata tggcctgcta ctatgggccc gagagcgtag gccacagaat
      721 aggagatgtc agctatggca tagacactgc catagactga tacgtggcgc acgtccacca
      781 gaaaggcgag cgtgggtagg agcgccgtgt ccaccaacgc gatgccgaag cagaggccgc
      841 agagcgagac cactaagggc gcgaaggatc ggcaggcagg tacgacgcaa gagctcactc
      901 cgattaccgc tagcccgaga gcgccataca gccactgcag gtgtggatag cgcgccgcta
      961 gacgcacggt gagatagacg cctaacacgt gtggcacgaa agccggcagc caaaccatgc
     1021 ccatctccca ctcggatgcc gccattgtgt gcttcatcca cgtggctatg gtgggctcga
     1081 gaaacgcaag gggaatgtta caagttgtga gcgctccggc tacaacagcg atgtaagggt
     1141 ctagcatgag gcgatggata ggtgtgccca cgggcaggtt ggcccgcgcc cgagccgcag
     1201 ctgagaaagg cttagccacc gccaggagta ggagtgcgtc gaagagcgac acggcggcga
     1261 gcactagaaa gggtacacgc ttgcccgcga actcgtagag gatgccccca aacggtggcg
     1321 ccactaggct tccaaagctg atgaaggcta gcgccacgcc cagagcacga ctgcgctcag
     1381 gctcctcggg atacttgtcg gcgatcatgg caatgccaga cgtgtccgcg aaggctgagc
     1441 ccaggccttg tagactgcgc gcagcgaaga gcgtggcata gtcttctgca aaggcaaaca
     1501 tgactgtgga ggcgaacatg acgcccaggc caataagcag cggcacgtcg tagctcatgc
     1561 gatcaatgaa aggcccgctt aaggggttca ccagaagctg caggatggcc ttggaggcaa
     1621 atagcacgcc tatcttcaca tcttcacttt ctgtggggta gcgaggccgc aggattgacc
     1681 gagccgaccc ggcagccgtc ggggacgccg aggtgttggc caagtaggcg ctggcattag
     1741 ccagagtggg cggcggcagg gtgggttccc acacctcaga gatcagggta gggctctcgc
     1801 tgcccccgcg catgtgagcg atatagtcgg gaacaatggg cgccggcgga gctcgggctg
     1861 ggcataaaag tcagggcaga gccatctatt gcttacattt gcttctggga tccagatctt
     1921 tcgaagctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     1981 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     2041 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
     2101 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc
     2161 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     2221 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     2281 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     2341 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     2401 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     2461 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     2521 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac
     2581 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     2641 ctcggcatgg acgagctgta caagtaagtc gacggcgcgc cgcggccgcg aattcgatat
     2701 cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2761 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2821 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     2881 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2941 tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3001 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3061 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3121 gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga
     3181 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     3241 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     3301 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     3361 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     3421 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3481 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3541 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3601 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     3661 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3721 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     3781 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3841 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     3901 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3961 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     4021 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     4081 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4141 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4201 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4261 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4321 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4381 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4441 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4501 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4561 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4621 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4681 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4741 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4801 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4861 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4921 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4981 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5041 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5101 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5161 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5221 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5281 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5341 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5401 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5461 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5521 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5581 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5641 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5701 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5761 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5821 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5881 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
