LOCUS       sequence-445278-        4822 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 4822)
  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 4822)
  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: HCT138_rAAV-eHGT_1505m-minBglobin-SYFP2-WPRE3-...
FEATURES             Location/Qualifiers
     source          1..4822
                     /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      834..843
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(840..842,843..845,846..1556)
                     /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(885..906)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1146..1165)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1493..1514
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1649..1669)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1856..1967
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2085..2225
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2085..2214
                     /label=AAV2 ITR
     rep_origin      2300..2755
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2387..2406)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2597..2618
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2772..2791)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2891..2913
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2951..2969)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3037..3141
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3142..3210,3211..4002)
                     /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(3360..3379)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4173..4761
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4662..4681
                     /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 gcgttgttgc tgagatgcag gttcaagcag gttgactgta
      181 aaatgtaggt gctgagaacc tactgggagc cgagctctca ttagctatac agctcaccat
      241 gtattctcag tatcgtgaaa agcagtaaag ttctacaaac acaccatcac cgtacacagg
      301 taacatttga gcctgagaac aagtgtaggg tacatgtatg atttcctctg cttcacatgc
      361 taggaaacag tctcggagga aagacgcgac ttgcccaagg cagcagagca attggtgagt
      421 caactgttgg acccagggag atcttgttcc aaagctggtt catttccatg gggctgtcgt
      481 cacacggaca ggaatagaaa cagactgcct gcctccttgc ccattttcct tccattaagt
      541 tatgaaagac cttggttttc taacctgtga ctgtaataca tagactttgc tttccatagt
      601 cactatttta ctgagcaatg gaattgtgat taaaaactat tacaaaatta atagtcaaat
      661 aagtacataa aataataaat aataaaagct tttaaaattt attttgtaac tgataaacta
      721 tttcatgaga gaaaagggag gagctcgggc tgggcataaa agtcagggca gagccatcta
      781 ttgcttacat ttgcttctgg gatccagatc tttcgaagct agcgctaccg gtcgccacca
      841 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
      901 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
      961 gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1021 tcgtgaccac cctgggctac ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc
     1081 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     1141 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1201 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1261 agctggagta caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg
     1321 gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg
     1381 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     1441 acctgagcta ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     1501 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag
     1561 tcgacggcgc gccgcggccg cgaattcgat atcataatca acctctggat tacaaaattt
     1621 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     1681 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     1741 ataaatcctg gttagttctt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     1801 ggacaggggc tcggctgttg ggcactgaca attccgtggc tcgagagatc ttcgactgtg
     1861 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     1921 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     1981 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2041 gacaatagca ggcatgagat ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     2101 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     2161 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     2221 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     2281 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2341 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     2401 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2461 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     2521 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2581 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     2641 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     2701 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     2761 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     2821 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     2881 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     2941 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     3001 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3061 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3121 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3181 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3241 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3301 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3361 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3421 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3481 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3541 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3601 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3661 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3721 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3781 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3841 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     3901 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     3961 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4021 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4081 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4141 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4201 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4261 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4321 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4381 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4441 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4501 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4561 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4621 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4681 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4741 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4801 tgctggcctt ttgctcacat gt
//
