LOCUS       sequence-445735-        5652 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 5652)
  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 5652)
  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: HCT46_rAAV-eHGT_1188m-minBglobin-SYFP2-WPRE3-B...
FEATURES             Location/Qualifiers
     source          1..5652
                     /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      1664..1673
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1670..1672,1673..1675,1676..2386)
                     /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(1715..1736)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1976..1995)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2323..2344
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2479..2499)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2686..2797
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2915..3055
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2915..3044
                     /label=AAV2 ITR
     rep_origin      3130..3585
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3217..3236)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3427..3448
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3602..3621)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3721..3743
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3781..3799)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3867..3971
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3972..4040,4041..4832)
                     /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(4190..4209)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5003..5591
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5492..5511
                     /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 gcgtcagaca tggtgctgga ggagctgaga gttctacatc
      181 ttggtccaca ggcaggcaga aggaggctgt aactgtgtgc agcttcagca taggcgacct
      241 cagagcatgc ctccaacaag gctatacctc ctaatagtgt cactccccat ccaagcattt
      301 aaacatgtaa gtctatgggg accataccta ttcaaaccac cacaagtacc agcctggtac
      361 ctgatcctta gtgtcattaa ctggggcagg gacctcagtc agacctgatc tttatccata
      421 tttgggttgt agggccagga gagtggctat aatgcctaaa ttaaggcctt gacatttccc
      481 gaagagccag tgtcgacaca gtgaagggga ctgaggggct tccctgtccg ttctgcatct
      541 gaaggagcta agactttcat gttccatctg tacatagacc tgcaacccgg catccatgct
      601 gtctttgtca acaggacctg aatcatccac acagccacag aaaggtggcc atggcatgca
      661 gcctgggctt tcccaaaaag gcttcagctc catggcccca aggccaaggc ttcagtccag
      721 ggcaacagcc tcaccagggc tgcagtacaa atccttctca gtggctagag gcatctggcc
      781 aagatgtctt ctttcatgac agatgaaaaa ctgtaaagaa aagtctcgac atctggaaaa
      841 cggtgccttt gttgtacagt taataagaga aagccatgtg ttaaaacaag atggctcttg
      901 tccgaacaat tttgaatgaa ctgctccaag acgtggatga gccaggagac aaactcagca
      961 gggagacaga gacttctgaa ttcagggaag tgctttctcc acttatatgt aagtaaaaac
     1021 atcatttcac ttagataatt aaggctcaga gcctttgtgc caggagtact cgcaggagtc
     1081 caggtgcctg ggtcccatcc agaccaatta aaccaaactc tggggaaggg gatgtaaggg
     1141 gagcaaagag catggggagc cttgcagggg ggaggggatg ggtggaagca tggaggtatg
     1201 ggagcaggca tctgtttata cttcggagat cactgatgct acactcacta aggttggaac
     1261 tttttccact gagcttacat tactataaca gtcttccttc tgtgactcca ggcacagggt
     1321 cctctccata cccagggcat ccaggaggct gaacctggcc atagagaccc agttgtaaat
     1381 gcatttctcc tctggatcct taagtccaag gctggcagca gaggtatgga tgctggtcac
     1441 aggatccctc catcccaggt ctggagattc tccaacctta ttactcagtg tacatgtgag
     1501 tgatgacctg ccaggcagct agatgccctg gccgagatgg tggcctggtc agtgtcaggc
     1561 aagagaactg gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat
     1621 ttgcttctgg gatccagatc tttcgaagct agcgctaccg gtcgccacca tggtgagcaa
     1681 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1741 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1801 cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1861 cctgggctac ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     1921 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1981 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2041 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2101 caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     2161 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
     2221 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     2281 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     2341 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag tcgacggcgc
     2401 gccgcggccg cgaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt
     2461 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2521 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2581 gttagttctt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2641 tcggctgttg ggcactgaca attccgtggc tcgagagatc ttcgactgtg ccttctagtt
     2701 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2761 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2821 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2881 ggcatgagat ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     2941 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     3001 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     3061 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     3121 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3181 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     3241 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3301 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     3361 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3421 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     3481 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     3541 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     3601 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     3661 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3721 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     3781 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     3841 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3901 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3961 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4021 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4081 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4141 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4201 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4261 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4321 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4381 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4441 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4501 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4561 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4621 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4681 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4741 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4801 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4861 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4921 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4981 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5041 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5101 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5161 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5221 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5281 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5341 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5401 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5461 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5521 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5581 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5641 ttgctcacat gt
//
