LOCUS       sequence-445285-        5151 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 5151)
  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 5151)
  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: HCT45_rAAV-eHGT_1187m-minBglobin-SYFP2-WPRE3-B...
FEATURES             Location/Qualifiers
     source          1..5151
                     /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      1163..1172
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1169..1171,1172..1174,1175..1885)
                     /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(1214..1235)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1475..1494)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1822..1843
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1978..1998)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2185..2296
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2414..2554
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2414..2543
                     /label=AAV2 ITR
     rep_origin      2629..3084
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2716..2735)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2926..2947
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3101..3120)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3220..3242
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3280..3298)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3366..3470
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3471..3539,3540..4331)
                     /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(3689..3708)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4502..5090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4991..5010
                     /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 gcgtgctatt gttcctggct gtgcacagct ctgtgctgcc
      181 cacaccccac actttttcag aggctctttg gctaaggaga aggtgggggt cggaacaagc
      241 cacttctcat tttctttaac cagcccaagg agaaacagag catggatttc tgagagtctc
      301 ataattaaat gaattaaaat gttttccttc gtggtgactt tgccaactcc catccacccc
      361 tctttggccc tgttcccaga gagtggcaga gctgggagga tgcagaatga catgttggct
      421 ccaaggggtg agcaggggtg cctccagagc aattagctta tgcatcatca ttctggggca
      481 ggggtgggga tttatggcta gccttcagag cattccagcc cagaccttct cagtccagat
      541 ctttttgctg gagacgttcc tggaagcaga agctgggtat tcattccaat cgcagtcaga
      601 tgggagccca ttcataagat agctgtccac aggctcctct cctgtcactc cagcctgacc
      661 ttgagccacc agcgctgtca aggaggaaaa cattgggctc aaccgcagcc ttcccggcag
      721 gcctaagtgg caacagagga agtcacgtag aagaatggga tgtgacggcc taatggctca
      781 gcatctctag tctgggtgtc atcctcaata tcctcttcac attcttctgc cagggaggtg
      841 acatgcaatg gattatgcct gtggtctgaa acctcccagg ccccagggcc accctttgtc
      901 ttagtttctg aagtggtctc tcttgttcta gagtttcagt ccaagtttgg gatgagccac
      961 tgaagctttc aaaggccagg ttaggtagga agccaaagcc caccacggtc cccttctgac
     1021 ctgaggccac aactcctggg actccctaat gagtttggct gccagtaacg agctcgggct
     1081 gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct
     1141 ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     1201 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1261 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac
     1321 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg
     1381 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1441 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1501 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1561 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1621 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     1681 catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1741 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga
     1801 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1861 tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc gaattcgata
     1921 tcataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     1981 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     2041 ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga
     2101 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2161 ttccgtggct cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2221 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2281 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2341 ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg
     2401 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     2461 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     2521 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     2581 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     2641 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2701 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2761 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2821 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     2881 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2941 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     3001 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3061 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     3121 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3181 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3241 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     3301 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     3361 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3421 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3481 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3541 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3601 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3661 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3721 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3781 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3841 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3901 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3961 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4021 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4081 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4141 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4201 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4261 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4321 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4381 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4441 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4501 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4561 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4621 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4681 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4741 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4801 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4861 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4921 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4981 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5041 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5101 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
