LOCUS       sequence-442621-        4941 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 4941)
  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 4941)
  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: HCT79_rAAV-eHGT_1306m-minBglobin-SYFP2-WPRE3-B...
FEATURES             Location/Qualifiers
     source          1..4941
                     /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      953..962
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(959..961,962..964,965..1675)
                     /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(1004..1025)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1265..1284)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1612..1633
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1768..1788)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1975..2086
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2204..2344
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2204..2333
                     /label=AAV2 ITR
     rep_origin      2419..2874
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2506..2525)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2716..2737
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2891..2910)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3010..3032
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3070..3088)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3156..3260
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3261..3329,3330..4121)
                     /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(3479..3498)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4292..4880
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4781..4800
                     /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 gcgtgcatta agatctgctt gcccagccgt ctagtgcaca
      181 cctttcatcc cagcactgag gaggcagagg caggtggatc tttgagttga ggtgagttcc
      241 aggacagcca ggatcacaca aagaaacact gtctcgaaaa accatccaac caacaacaaa
      301 gtcctcctcc caccgccaag aaaaagatgt gtttgattac agcagagact gacaaggtgg
      361 cggcatgaca agagctgcgg gttgctgcac tttctgtctc agtgttggcc tgtggcgcac
      421 ttcccaggac tgcaggagat aaagtgtact ttcaaatctt tatataagac aaaagtcacc
      481 aaggccactg aattatgact gtagaaaaaa gtcaccactt ccaatgtgct gctctaagga
      541 aaaaggaagc aataaaattt gatcgattca acttttaagt acacatgagg ctcaggaaat
      601 ataattccta agccataatg ttgaattaac atatctcatt acacctaatt tcttacagtg
      661 gtgcaatgtg tgatgttcca gacacaagga caagaaagcc acatgaagta agaatacgct
      721 aacaaagcca actgtctttc cagattccta gtattaaatg ttgccttcct tagtcacaaa
      781 accaatctaa catttcaata cgaaggaaaa tcaagccaaa gtcacaggag ggacaagtgc
      841 tgtctcaggt aagcacacgg agctcgggct gggcataaaa gtcagggcag agccatctat
      901 tgcttacatt tgcttctggg atccagatct ttcgaagcta gcgctaccgg tcgccaccat
      961 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1021 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1081 caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1141 cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca
     1201 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1261 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1321 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1381 gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg
     1441 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga
     1501 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1561 cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1621 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagt
     1681 cgacggcgcg ccgcggccgc gaattcgata tcataatcaa cctctggatt acaaaatttg
     1741 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1801 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     1861 taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     1921 gacaggggct cggctgttgg gcactgacaa ttccgtggct cgagagatct tcgactgtgc
     1981 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     2041 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     2101 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     2161 acaatagcag gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     2221 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     2281 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     2341 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     2401 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2461 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     2521 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2581 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     2641 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2701 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     2761 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     2821 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     2881 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     2941 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3001 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3061 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     3121 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3181 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3241 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3301 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3361 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3421 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3481 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3541 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3601 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3661 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3721 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3781 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     3841 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3901 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3961 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4021 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4081 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4141 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4201 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4261 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4321 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4381 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4441 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4501 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4561 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4621 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4681 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4741 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4801 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4861 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4921 gctggccttt tgctcacatg t
//
