LOCUS       sequence-445273-        5010 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 5010)
  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 5010)
  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: HCT75_rAAV-eHGT_1302m-minBglobin-SYFP2-WPRE3-B...
FEATURES             Location/Qualifiers
     source          1..5010
                     /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      1022..1031
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1028..1030,1031..1033,1034..1744)
                     /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(1073..1094)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1334..1353)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1681..1702
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1837..1857)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2044..2155
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2273..2413
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2273..2402
                     /label=AAV2 ITR
     rep_origin      2488..2943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2575..2594)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2785..2806
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2960..2979)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3079..3101
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3139..3157)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3225..3329
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3330..3398,3399..4190)
                     /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(3548..3567)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4361..4949
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4850..4869
                     /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 gcgtggtccc cagattagca ttagaacaca aagagcatta
      181 ggccaaccaa ctacaagcaa gtacctaaaa tgtggaagca gatttaaaat ttagtaatta
      241 aatggactga agagtttgaa taagtaaatg agaaaaagta tagttttctg tgaatggatt
      301 acaaagtgat gtggcatctg gtggaattaa aacttagtag tctttaaact atgtgtgtgt
      361 gtatgtgtgt ccactgcttt gcatttttaa ttcttggtgg agagtcattt ttttaaaggg
      421 ctcatttaaa aatagaagat gcattgattt gctttgatat ccagtttttg ttattggagt
      481 gaatgcaatt atgatggact aagtgcccaa gtaatgagca atttggcaga gacagactgt
      541 cctaaagcta tggaaacaag ctatgcttgt gtctgcaggt cagatcagaa catcagcgga
      601 gcgatgtgga tgggagaaaa ttacgcaaca aataagattt ttttccatga aaggaaaaac
      661 aaggtagaag agatgaagaa tcttttagag actcttatca tatacagtct tgctatatgg
      721 taatcgaagg gccaatatag gcaagtagta gaaacagcag agcctagaca gatattgttt
      781 caaaatacat tctgtcttag gctcaaagta aaagaaacct gtctagaaag agattcaagg
      841 ggaagttccg tgaaagtaaa aatgtgggtg agattttaaa gaagtctaat ttgtcctcaa
      901 atactaagta gatctgttgc tagtcaagga gctcgggctg ggcataaaag tcagggcaga
      961 gccatctatt gcttacattt gcttctggga tccagatctt tcgaagctag cgctaccggt
     1021 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1081 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1141 cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg
     1201 gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct accccgacca
     1261 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1321 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1381 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1441 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca
     1501 gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca
     1561 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1621 caaccactac ctgagctacc agtccaagct gagcaaagac cccaacgaga agcgcgatca
     1681 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1741 caagtaagtc gacggcgcgc cgcggccgcg aattcgatat cataatcaac ctctggatta
     1801 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     1861 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     1921 ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc
     1981 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc gagagatctt
     2041 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2101 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2161 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2221 attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc gcaggaaccc
     2281 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     2341 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     2401 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     2461 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     2521 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     2581 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2641 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2701 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2761 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     2821 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     2881 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     2941 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     3001 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3061 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3121 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     3181 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3241 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3301 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3361 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3421 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3481 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3541 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3601 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3661 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3721 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3781 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3841 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     3901 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     3961 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4021 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4081 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4141 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4201 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4261 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4321 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4381 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4441 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4501 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4561 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4621 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4681 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4741 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4801 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4861 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4921 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4981 tggccttttg ctggcctttt gctcacatgt
//
