LOCUS       sequence-445759-        5587 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 5587)
  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 5587)
  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: HCT135_rAAV-eHGT_1502m-minBglobin-SYFP2-WPRE3-...
FEATURES             Location/Qualifiers
     source          1..5587
                     /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      1599..1608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1605..1607,1608..1610,1611..2321)
                     /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(1650..1671)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1911..1930)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2258..2279
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2414..2434)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2621..2732
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2850..2990
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2850..2979
                     /label=AAV2 ITR
     rep_origin      3065..3520
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3152..3171)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3362..3383
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3537..3556)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3656..3678
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3716..3734)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3802..3906
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3907..3975,3976..4767)
                     /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(4125..4144)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4938..5526
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5427..5446
                     /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 gcgtttaggg tttgggcatt aatatttttt tctaagaaaa
      181 aaaaaagtct ttgctatggt ctatggtcta gatatatgtt cctcaaaggt tcctgttccc
      241 ttatacctcc agggcagtgg tgtaagcttt aagaggtagg gtctaacaga agttaattaa
      301 atcccaaacg tccacactga taaaaagagt cccatctgtc tttgaagagt gggtagttat
      361 aacgggaaac taccccaaga gtatggccat ccctttctgc ttcttcacac aggcagggct
      421 gtgtgctggc gccaagcttc ttggagcccc caccaccaga atcttgaact tcagtacaca
      481 gcacacaggt attttgttaa agcaaccatg agtgggcaga ggcagtgtcc cagaaggctc
      541 ctgtgtcaga ctggctgagg ccttctgctt ggttagtcta acccgctgac agccagccct
      601 gagaaagaat tgtgtacaga atccattaca agtggtttca tttcctctgt gattcacaaa
      661 aaagctgaaa gttatagaat ttcaaacgta tgttgcatct cctaaactac ccattttgtg
      721 agaaatcaag cccaaagctt tgcatgtctt ggaagctata cctgaggtga agttagtcta
      781 agtaacaagt ctcagtcatt gtgaagtatt ccgatgcacg gtttgtcagg tcattgttct
      841 ttcctcttgt aaatactacc cccaagcccc caaaagtcac ctatctgaaa ggtaaaatgt
      901 atttctaaag ctcaaggcat ttattctgca aggggtgggg gtggagtggg gttgaaaaaa
      961 aaagagaagc catgtgttta taaaaggttg gggcagggga tgagggaagt tctgcggtaa
     1021 tgacagggga gttccagagg aagttatggg ctctctctcc tctctcatga gaaaaaaaag
     1081 atagagcggt aaacagctta gctgttaaca catctccaaa tctacccagc cagttatctt
     1141 ccttgcacac tcctctttcc ccttcaccac ctctctcctt gttttccgtt tgggcatctg
     1201 aaatttaaaa ggggaaacag actttagtag tagtaaattc ctaagctaaa tgctcagaaa
     1261 atagcttagt atatctttga tgagggatga ttatgcttcc cgccccctag ggagccgcag
     1321 agtggatttg aaacaaagta tcttttagtt acaagtgagt gaagggttta gagatgattt
     1381 ctgatttagt cacttttctc cctggctggt acctaagtaa cagcctggca gaagcgacct
     1441 tagggagaag ggttcatttt gtttcaccat tggagaggac acaggagtcc acagtggttg
     1501 ggaaggagct cgggctgggc ataaaagtca gggcagagcc atctattgct tacatttgct
     1561 tctgggatcc agatctttcg aagctagcgc taccggtcgc caccatggtg agcaagggcg
     1621 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1681 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     1741 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
     1801 gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     1861 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     1921 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1981 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2041 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     2101 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     2161 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     2221 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     2281 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagtcgac ggcgcgccgc
     2341 ggccgcgaat tcgatatcat aatcaacctc tggattacaa aatttgtgaa agattgactg
     2401 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2461 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag
     2521 ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     2581 tgttgggcac tgacaattcc gtggctcgag agatcttcga ctgtgccttc tagttgccag
     2641 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2701 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2761 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2821 gagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc
     2881 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     2941 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     3001 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     3061 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3121 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     3181 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3241 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     3301 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3361 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     3421 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     3481 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     3541 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     3601 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     3661 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     3721 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     3781 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3841 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3901 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3961 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4021 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4081 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4141 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4201 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4261 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4321 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4381 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     4441 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     4501 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     4561 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     4621 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     4681 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4741 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4801 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4861 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4921 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4981 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5041 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5101 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5161 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5221 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5281 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5341 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5401 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5461 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5521 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5581 cacatgt
//
