LOCUS       sequence-445736-        6310 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 6310)
  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 6310)
  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: HCT80_rAAV-eHGT_1307m-minBglobin-SYFP2-WPRE3-B...
FEATURES             Location/Qualifiers
     source          1..6310
                     /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      2322..2331
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2328..2330,2331..2333,2334..3044)
                     /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(2373..2394)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2634..2653)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2981..3002
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3137..3157)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3344..3455
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3573..3713
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3573..3702
                     /label=AAV2 ITR
     rep_origin      3788..4243
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3875..3894)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4085..4106
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4260..4279)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4379..4401
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4439..4457)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4525..4629
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4630..4698,4699..5490)
                     /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(4848..4867)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5661..6249
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6150..6169
                     /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 gcgtatcact gactagttac atgtctttaa taacagtaaa
      181 cccactgctc agttaatcca atatactctg gatgctcaag agatgagaat ggttcactgt
      241 aactggttaa aataacttag aatgtcctgt aaaaagtaga gtttaatctc ttagccattc
      301 ttcctgcaat tccagacact accagcaggg gtcagtcgga gattctttag aacatcatct
      361 gtaagtttaa gtactaatta actctacatg ttatctgtac ttaaaacacc ttatcagaat
      421 aataagataa aaaagtaaca gttacacaat attcaagctc cattcgctac aatcaacatg
      481 agattctttc aatacgaccc tttaaataag tttagcaagc aaatgctagc tgaatgtcta
      541 ttcatgcctt atacttctgt aggagctaca gagcaaagat tcccttcaag gatgtcaata
      601 aatacttcct ttagatctag tcgtcccttc gactagagga gactttaaga agctctcatc
      661 gctaactggt ttctggatcc cctgtgccca accatcagaa agcctttagt tctttccttc
      721 taaatattag gacagagatg gattctttaa aatctctact tttcccccct ctctctgtaa
      781 tcttgtccaa atcttgcaat agaaaagatt cacagagaaa ggattttaaa aatcgggaaa
      841 gcaccgagga gggtggcaag agtggtttag gaggctctga ggaacgatgt gttgtgcata
      901 ggctaccctt tagctgatcc actagatctg cctgactttg cagtcatggg taaataggct
      961 tcacctattg ccagtccctt ctctgtgaac aaagagtacc ttccctgagg gcagcatgta
     1021 ttctggctcc aaagccacac tctgagacat gtttctctct gtcttcaaaa cacatcccca
     1081 ctaatttttt tttcttttag aaaatggcac atttttgcac agactctaac tttactagag
     1141 gtcaggataa aagccttcct atttttggga atctctcatg attgacccaa aggaccaatc
     1201 tcaattcttt ctatgccctc acacctttat tttaacccaa agaggttcta aactgtctgt
     1261 ttatcaacag attgagttca ttgctcagtg ggaatgtggt agttaaccat ttaatggcaa
     1321 atgaggttgt ggtttaatgc tatcattttt cccgtagaat catgcaattt tgaatatttt
     1381 tgagccaagt ggatgagtga ttttctgtat tttacttaca tctttcctta ttgttctgcc
     1441 ttttatttgg aaacagtgat tgctttctgc catttgctta atgagtttga cgtcatcccg
     1501 ttgggcatca gattagagtc ataaaaggaa caaaaagtca cattgctcta tttctagtgc
     1561 tataaacaaa attgcatgaa aacgaaggcc aaaaaaataa gtatttgaaa atgcaaatgc
     1621 ttttcagaat gaaaaccaaa ttccagagat gctgctctgt tctttccaat ttgtgtcatt
     1681 aaattttgat tctgtaggaa aagagtagac tgaagtagac aacaacagac aactgcttta
     1741 tagcccgtta gtgtcaacgt tccctgcctg catgcacgtc ctcccaaata tgggaattaa
     1801 ttatcacaga ctgtgggcca agcagatggg caactctgat aaaggaataa aactttgggg
     1861 tgaacaaata gatgtgacaa accactttac tgtaagacat cccctgcaaa taatgccgtt
     1921 caatagactc ctcataggat ggttctgtgg ctattggacg aataaacaga gactgagcca
     1981 tcccatgatc aaggctctgg cataccacac catagtgatg tgcctgccat tatgttgcaa
     2041 ttcttctacg ccctaaactt ctagcattac agctggcctc tcgggtctgt gcttcccacc
     2101 tctctgaatt ctgcatctgt ggattcaatc aactgctaac gaaaaatgtt ggggagggga
     2161 aaaaaatcta tctatactga aaaagtgcat cgttcctgtt actattgcct gtatattact
     2221 tccatggcga gctcgggctg ggcataaaag tcagggcaga gccatctatt gcttacattt
     2281 gcttctggga tccagatctt tcgaagctag cgctaccggt cgccaccatg gtgagcaagg
     2341 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     2401 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     2461 tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     2521 tgggctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     2581 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     2641 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     2701 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2761 actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca
     2821 acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc
     2881 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     2941 agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     3001 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagtc gacggcgcgc
     3061 cgcggccgcg aattcgatat cataatcaac ctctggatta caaaatttgt gaaagattga
     3121 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     3181 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     3241 tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3301 ggctgttggg cactgacaat tccgtggctc gagagatctt cgactgtgcc ttctagttgc
     3361 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3421 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3481 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3541 catgagatct cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     3601 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     3661 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     3721 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     3781 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3841 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     3901 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3961 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     4021 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4081 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     4141 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     4201 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     4261 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     4321 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4381 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4441 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4501 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4561 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4621 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4681 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4741 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4801 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4861 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4921 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4981 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5041 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5101 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5161 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5221 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5281 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5341 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5401 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5461 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5521 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5581 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5641 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5701 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5761 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5821 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5881 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5941 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6001 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6061 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6121 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6181 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6241 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6301 gctcacatgt
//
