LOCUS       sequence-445763-        5094 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 5094)
  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 5094)
  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: HCT134_rAAV-eHGT_1501m-minBglobin-SYFP2-WPRE3-...
FEATURES             Location/Qualifiers
     source          1..5094
                     /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      1106..1115
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1112..1114,1115..1117,1118..1828)
                     /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(1157..1178)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1418..1437)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1765..1786
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1921..1941)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2128..2239
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2357..2497
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2357..2486
                     /label=AAV2 ITR
     rep_origin      2572..3027
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2659..2678)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2869..2890
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3044..3063)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3163..3185
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3223..3241)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3309..3413
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3414..3482,3483..4274)
                     /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(3632..3651)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4445..5033
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4934..4953
                     /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 gcgtggtcct tgtaagtcct gtaatattaa tttgtgttga
      181 tcctagtcac aatgaatgtt gtcagtcagt agactgacag tctagttagt tcctatccct
      241 ctgtagtatg agggctgggc tgtctaaaca taaccagcca tcctggccaa cttctggctg
      301 ggagtgatga actcccgggg gcccttcctg tggcataaag tccgattccc caggttttcc
      361 ctgtttccca aaggccagaa ctttttttta aagaagaatt ttaggaagcc tcagtttcac
      421 aaccctaatt tttacaatgg aaactttttt gtttcctggg gcccagtgaa accttaattt
      481 taataggtca cagtgcagga gacagtccat ttggcaacag tgtctgtggc ttctgctttc
      541 tcgacctgga tgtactgaat ccatggtgta atgccagttg tcttcacagc cataggagta
      601 gaaagaatca ctgtgtaggg tcctgtccaa gttgttttca gtgcccttta gggcagccac
      661 ttgacttgaa caaatagaaa ctggtggtga acttggaggt tggctgggtc tctcagctag
      721 tccaagactg agggtctata gtgactggac agaggaatag tcagagagtt ctgtcaactg
      781 ctggtctctg agctcgagaa acagtggggg aggtcttcca aatataatct catagggagt
      841 aagtccctcc ccatatgggg tgaaacaggt ctatgcaaag caaaggaagg aggcctatcc
      901 acccctctca ttggacacta atgagagttt tgttagggtt tctttttatg tcctggttat
      961 tcttttgacc tggccacaac tctgaggtct gtatgcacag tgaagttttc aggagctcgg
     1021 gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga
     1081 tctttcgaag ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac
     1141 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1201 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac
     1261 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca
     1321 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1381 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1441 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1501 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     1561 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
     1621 caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1681 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa
     1741 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1801 cactctcggc atggacgagc tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg
     1861 atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     1921 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     1981 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc
     2041 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2101 caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt
     2161 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2221 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2281 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg
     2341 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     2401 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     2461 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     2521 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     2581 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2641 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2701 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2761 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     2821 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2881 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     2941 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3001 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     3061 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3121 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3181 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3241 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3301 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3361 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3421 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3481 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3541 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3601 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3661 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3721 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3781 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3841 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3901 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3961 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4021 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4081 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4141 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4201 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4261 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4321 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4381 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4441 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4501 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4561 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4621 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4681 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4741 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4801 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4861 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4921 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4981 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5041 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
