LOCUS       sequence-445760-        5018 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 5018)
  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 5018)
  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: HCT128_rAAV-eHGT_1495m-minBglobin-SYFP2-WPRE3-...
FEATURES             Location/Qualifiers
     source          1..5018
                     /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      1030..1039
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1036..1038,1039..1041,1042..1752)
                     /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(1081..1102)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1342..1361)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1689..1710
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1845..1865)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2052..2163
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2281..2421
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2281..2410
                     /label=AAV2 ITR
     rep_origin      2496..2951
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2583..2602)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2793..2814
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2968..2987)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3087..3109
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3147..3165)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3233..3337
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3338..3406,3407..4198)
                     /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(3556..3575)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4369..4957
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4858..4877
                     /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 gcgtaaaaca ccttaaccag gggcagtgtg atgggatcag
      181 gttactcccg agactttaac ggatgcagtt gggctggggg gtaggcattc caggaagaat
      241 tcaacatggt gttggaacgg ataaaaacaa gcagctgaga gtcatagcag ccaccccgag
      301 gttttaagct gaggagatag gaaataccca gagagggctg cacatctcca ttcttcggac
      361 tgtccctaga tgcatttggt cccagaaagc agctgctatg actgactgct ggagaaaaag
      421 gtcttgatgg tatggaattc aggcgactgt aattacagat aataggtctc aatttgcccg
      481 tcaggaattt gcacagttgg ctctatcact gggaatttaa atatttttac ttcccaattg
      541 tgcattgaag tcagacagca gatgaagcag aagtggtcca agattctcac cagctcagta
      601 catttgcctt ggaaatgtcc agaacatcct tgtggccttc ccccatcaag ggagttttcc
      661 ctgagcaagc tcaagccact gttgggtcct gggtctggaa agttgaagaa gcagggcttg
      721 ctggcatggt aacaatagag cattttcttg gagcctgtga tgaattatga gaactgtgtc
      781 cactggatgc tagtctttga ttgaggatga catagtgttc catcacactg tctggaacca
      841 gttccaaaag aataaaaaag tgacttgtaa atgaatgttg gaagcatttt cttccttcct
      901 tccttccttc cttccttcct tccttccttc ctccttgagc tcgggctggg cataaaagtc
      961 agggcagagc catctattgc ttacatttgc ttctgggatc cagatctttc gaagctagcg
     1021 ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1081 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1141 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
     1201 gtgccctggc ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt cgcccgctac
     1261 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1321 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1381 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1441 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
     1501 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     1561 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1621 ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag
     1681 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1741 gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa ttcgatatca taatcaacct
     1801 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     1861 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     1921 attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc
     1981 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggctcga
     2041 gagatcttcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2101 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2161 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2221 gggggaggat tgggaagaca atagcaggca tgagatctca cgtgcggacc gagcggccgc
     2281 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     2341 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     2401 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     2461 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     2521 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     2581 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2641 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2701 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2761 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2821 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     2881 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     2941 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3001 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3061 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3121 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     3181 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3241 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3301 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3361 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3421 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3481 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3541 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3601 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3661 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3721 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3781 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3841 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     3901 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     3961 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4021 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4081 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4141 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4201 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4261 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4321 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4381 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4441 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4501 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4561 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4621 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4681 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4741 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4801 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4861 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4921 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4981 acggttcctg gccttttgct ggccttttgc tcacatgt
//
