LOCUS       sequence-445264-        5296 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 5296)
  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 5296)
  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: HCT43_rAAV-eHGT_1185m-minBglobin-SYFP2-WPRE3-B...
FEATURES             Location/Qualifiers
     source          1..5296
                     /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      1308..1317
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1314..1316,1317..1319,1320..2030)
                     /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(1359..1380)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1620..1639)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1967..1988
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2123..2143)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2330..2441
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2559..2699
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2559..2688
                     /label=AAV2 ITR
     rep_origin      2774..3229
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2861..2880)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3071..3092
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3246..3265)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3365..3387
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3425..3443)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3511..3615
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3616..3684,3685..4476)
                     /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(3834..3853)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4647..5235
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5136..5155
                     /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 gcgttgtaag agtaataatc gccggcgggt gtcaagtctc
      181 agttggtact tttcatatat ttctgtgctc atctcttcag aatacaatca tacaattagg
      241 tttggagtca aactcttaga actcccctgt acttgctgta tacaccctgt tagacagcag
      301 tcaccagatc agaaactatt tctgtagtgt agatttccaa acagaatgat ttaaccactg
      361 ccagatcact aagcagagtc ctcctcaggt gctgtgtcta gattggaagg ctccccagag
      421 caggtgtccc taaagcaggt gctcttgtgc tctctggtgg cctggtgttc agcagcttag
      481 ccatcttcct ttccctttgt tcctttctta tacagtggcc ctgtgataaa aagcctttgg
      541 agctctcttt aaaaataata atctccatgc agtttggaat acttcagcta attaggaatt
      601 ttcaaatcct gccaagattg agaaagagct cagctcaaaa agtattgtct aactcactca
      661 tggattgatc caatagcagt attttaacct gggctctgaa atattttttc caactattat
      721 tttagtgcaa atgaaaattt tgaaaaatat gttttggcta aatgatgtat cactgtatct
      781 cagaactttg ttactaagct cctagcaatg aatctaataa tagccaagaa cagttctgtg
      841 aatttaaaat aaaatctaaa atatctattt catctctgta aagaatttgt gtgcatgtga
      901 gtgtggtatt caatttcaaa gaactacatt ttgttaaaat gtatgtaatt aattctctgt
      961 gtttatttta gaatataagt ataatggagt tttattataa ataagttttg aaaatttgcc
     1021 tattaccaac aaataaaagg aaaagacaca agatagctat agaggaaata cctttgctat
     1081 tataaaaata tttaaggagt tcaatattta ttaccaatat ttttatttaa ttctttatat
     1141 aatacacctt ctcaaaatcc aataaataat tatacaataa ataattatac aaaggccata
     1201 aggaaccgcc ggcggagctc gggctgggca taaaagtcag ggcagagcca tctattgctt
     1261 acatttgctt ctgggatcca gatctttcga agctagcgct accggtcgcc accatggtga
     1321 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     1381 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     1441 tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     1501 ccaccctggg ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
     1561 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     1621 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1681 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1741 agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca
     1801 aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact
     1861 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1921 gctaccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1981 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taagtcgacg
     2041 gcgcgccgcg gccgcgaatt cgatatcata atcaacctct ggattacaaa atttgtgaaa
     2101 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2161 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2221 cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     2281 gggctcggct gttgggcact gacaattccg tggctcgaga gatcttcgac tgtgccttct
     2341 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2401 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2461 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2521 agcaggcatg agatctcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt
     2581 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     2641 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     2701 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     2761 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2821 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     2881 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2941 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     3001 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3061 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     3121 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     3181 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     3241 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     3301 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3361 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     3421 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     3481 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3541 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3601 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3661 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3721 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3781 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3841 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3901 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3961 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4021 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4081 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4141 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4201 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4261 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4321 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4381 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4441 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4501 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4561 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4621 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4681 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4741 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4801 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4861 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4921 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4981 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5041 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5101 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5161 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5221 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5281 ccttttgctc acatgt
//
