LOCUS       sequence-446469-        5350 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 5350)
  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 5350)
  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: HCT39_rAAV-eHGT_1181m-minBglobin-SYFP2-WPRE3-B...
FEATURES             Location/Qualifiers
     source          1..5350
                     /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      1362..1371
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1368..1370,1371..1373,1374..2084)
                     /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(1413..1434)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1674..1693)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2021..2042
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2177..2197)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2384..2495
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2613..2753
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2613..2742
                     /label=AAV2 ITR
     rep_origin      2828..3283
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2915..2934)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3125..3146
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3300..3319)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3419..3441
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3479..3497)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3565..3669
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3670..3738,3739..4530)
                     /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(3888..3907)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4701..5289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5190..5209
                     /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 gcgtgagact ctgaaactct tggtactttt tctcatggtc
      181 cagttttcat cacttttaca caattcttta ttataagcca ctagcgagca cctctgacat
      241 tcactgaaaa ctaaacacta aacacaaagc aaaagcaaaa caagtgctcc aacacaaaat
      301 caagatggct gaccagcacc aacgatatat gatgggcctt catctatagc ctgtgttcct
      361 tcagtattgt ataatggcca cctctgacct tttaattttt ggcaatcaat gggagaatgt
      421 tactcataaa cctgacattg tttcctacac ttgggtgccc ccgatgcctg agttgcacat
      481 tccttctacc ctcagtgaga acctctgccc aggagtccag actccaggtc cctgctgtcc
      541 aggtcccacc caggccaacc tcagcttcac atatgcaccc acccacccac ccaccaggcc
      601 caccccactc catgtaagag ctaattatct gtaagtgctt tcagcagcca tttccttagg
      661 aatgtgtaag tgctttgatt agagtactct cttggcacca actttatggc tagcaatggg
      721 aagcgggcaa ttaggggctg gagacagtcc tgaaattgga ctagtgtttg aaggagctgg
      781 cctcaggcag gactgctcct cagcctttca taggagctga gctgagtgac ctcctttgac
      841 ctggccattt attccagctc ctcccatttt ttctcaaagg ctccatagag ggaggcatag
      901 gcctggagat cagaaacaag gaccagagtt agcctgctgt cctgtgtggg cggaccaatc
      961 accttgattc tctgtttcct catcgttcct cctacactgg agtctgtctg ttgaattctc
     1021 tgctgggcat caaggacctg gagagaattc agtgcagggt ttaccctgag ggcaatatga
     1081 atgtaaccta gacagaccat accaaggatg cacgctgcaa ctgtgatcag atcacagtgg
     1141 tatggcagct gcaaggacag aaatggcctc tgtctgggga agtaggtgac cttacctggg
     1201 tctgaaacag gaggtgagag tgaaatgagg acaacacttc agctttaagt aaattaggtg
     1261 tgaggcgcga gctcgggctg ggcataaaag tcagggcaga gccatctatt gcttacattt
     1321 gcttctggga tccagatctt tcgaagctag cgctaccggt cgccaccatg gtgagcaagg
     1381 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1441 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1501 tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     1561 tgggctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     1621 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1681 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1741 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1801 actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca
     1861 acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc
     1921 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     1981 agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     2041 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagtc gacggcgcgc
     2101 cgcggccgcg aattcgatat cataatcaac ctctggatta caaaatttgt gaaagattga
     2161 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     2221 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     2281 tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     2341 ggctgttggg cactgacaat tccgtggctc gagagatctt cgactgtgcc ttctagttgc
     2401 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2461 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2521 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2581 catgagatct cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     2641 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     2701 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     2761 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     2821 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2881 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     2941 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3001 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     3061 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3121 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     3181 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     3241 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     3301 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     3361 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3421 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     3481 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     3541 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3601 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3661 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3721 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3781 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3841 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3901 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3961 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4021 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4081 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4141 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4201 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4261 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4321 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4381 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4441 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4501 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4561 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4621 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4681 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4741 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4801 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4861 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4921 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4981 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5041 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5101 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5161 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5221 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5281 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5341 gctcacatgt
//
