LOCUS       sequence-446468-        5702 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 5702)
  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 5702)
  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: 
            HCT55_rAAV-eHGT453m_eHGT_1137m-minBGprom-SYFP2-WPRE3-bGHpA
FEATURES             Location/Qualifiers
     source          1..5702
                     /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      1714..1723
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1720..1722,1723..1725,1726..2436)
                     /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(1765..1786)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2026..2045)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2373..2394
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2529..2549)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2736..2847
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2965..3105
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2965..3094
                     /label=AAV2 ITR
     rep_origin      3180..3635
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3267..3286)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3477..3498
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3652..3671)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3771..3793
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3831..3849)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3917..4021
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4022..4090,4091..4882)
                     /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(4240..4259)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5053..5641
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5542..5561
                     /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 gcgtcgcgtt tcacaaatca gtaagaatta cactccaaaa
      181 ataaaacagg gaagggtatt agtatgtgat acgcatggat aaagtagaag aggccaaaaa
      241 tacaaagaac cgtaaactca ctagtgatgg aagaaacgct aattaaaata atgagctttc
      301 ctgacccttt atcacatggg cacattttca aaggacaaga ctctgctgag gagaacacag
      361 aacaggatcc ttcagacaag cttcagcaac caattttagg atgttcttaa atagatcgat
      421 tcttctactt tttttaaaaa taacatagat ctgaaaagct ttataacttt tggttttttt
      481 ttaaggacca tattaacagc atcttccagt ttttctagag taaatgaata tgtatccctt
      541 ttataattaa caatatacat atattcattt tgtaaaaaga ctataaaaat agaagagaaa
      601 atgaagagat gtttatttct tttacacgca tcagactaag gccttcagct cagcaggggg
      661 cagagtttat catctgtact ctctgaagtg tcacactgcc ctgcctttgt gtgcccttgc
      721 ctgtccctct gtgacggtca ccctttcggt tctggcttgg agcctggtca tcagtgcctg
      781 gcggcctgtg cctgggctgt gctgagcaca tttggtcgtt gtcctagtga tgtgacccta
      841 ccttagactt ccccatctct tgaggtggct aggtaccggg agtctggaga gagggtggga
      901 gcagccattc tgcagcagtg ccttcttggg gtcatgggtc tgtaggtgct gctgtggagg
      961 gagagatcag cctattctgg cttcatttct gagctgcaaa ctgcctgggt gtctggagaa
     1021 gcaggttggc gtggtggtta gcagtgcgtg ggcggggttg cccgctcttg atttatgatt
     1081 tctttgtctc tgtggaagca cttaagtgca ggctttagtt ccaatgacac tcaggagcct
     1141 ctggattcca gcactgggga tgggggtggg gtagaacgtt ctcaggcctc accaacccct
     1201 cccctgtgtg ctgcctttgg gagagtccca aggcttcagc attacttaat taattaggcc
     1261 tctactgcta cataggctca gattcaaaag aacagagtgg cccacgtcag ccattcccgg
     1321 aaaagtctga tggctggaag ccagaggact atgtgtctgc cttgctgccc ttggccagcc
     1381 catcctgaat gcccagactc ggacaatgga gtaggtacag aagggtaaag acagtgtctt
     1441 ctgtaccagt aagtgggccc tgatctgctc tctacagctt ccagagaaag ggcctggcca
     1501 atgagcggcc ttttgagtag cagatacctc acatgcattc tgatagaaag cctggcccca
     1561 gatcactgtg actttagccc tcaggtttct tttgcacttc ctatttcact gagtttgagc
     1621 gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg
     1681 gatccagatc tttcgaagct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag
     1741 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1801 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg
     1861 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac
     1921 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1981 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     2041 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2101 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     2161 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     2221 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     2281 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag
     2341 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2401 gccgggatca ctctcggcat ggacgagctg tacaagtaag tcgacggcgc gccgcggccg
     2461 cgaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2521 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2581 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt
     2641 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     2701 ggcactgaca attccgtggc tcgagagatc ttcgactgtg ccttctagtt gccagccatc
     2761 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2821 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2881 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat
     2941 ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     3001 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     3061 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     3121 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     3181 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3241 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3301 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3361 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     3421 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3481 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     3541 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3601 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     3661 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     3721 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     3781 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     3841 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     3901 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3961 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4021 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4081 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4141 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4201 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4261 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4321 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4381 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4441 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4501 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4561 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     4621 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4681 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4741 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4801 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4861 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     4921 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4981 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5041 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5101 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5161 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5221 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5281 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5341 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5401 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5461 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5521 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5581 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5641 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5701 gt
//
