LOCUS       sequence-445279-        5084 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 5084)
  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 5084)
  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: HCT41_rAAV-eHGT_1183m-minBglobin-SYFP2-WPRE3-B...
FEATURES             Location/Qualifiers
     source          1..5084
                     /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      1096..1105
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1102..1104,1105..1107,1108..1818)
                     /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(1147..1168)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1408..1427)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1755..1776
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1911..1931)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2118..2229
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2347..2487
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2347..2476
                     /label=AAV2 ITR
     rep_origin      2562..3017
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2649..2668)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2859..2880
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3034..3053)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3153..3175
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3213..3231)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3299..3403
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3404..3472,3473..4264)
                     /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(3622..3641)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4435..5023
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4924..4943
                     /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 gcgtccatga aatgcagagg ttaccatctg tgggcgagag
      181 gaaggagaac tgagagactg ggaagagtct tagtccactg agaagggaaa ttagtagacg
      241 tcagtctctc ctagttgcgt actgggtgat gcaagactgt tctaagaatg cagatgcaga
      301 tatcccttgg gtgggtagga gaggagatga atccaggatg ttttagaaat tcagaaaaac
      361 taaacaaagc taggaattag aaacaacttc aatgccagag taagtcagta cgtgagtggt
      421 gttgacagca acagaaattg agattggaag gagccagatc cttcttgggt tgggcttctg
      481 aagttctttt aggaattcca attgagagag tccaaaggat ctagctggta agagaaaaat
      541 gtgtctggca gagtcccagc tccaggctct catttcagat gcgtaatttt atatttacat
      601 attgaaaagt tcatttattt tggtgtgtgt tttgggagca agtccttgag ccatggagtc
      661 agcacaatga ggacctaaag caattacatc acttgccaaa tcctccagtt tcccttagct
      721 ctcagctgct ccctcaccca cagtctcggc tgatctagat ctacgggctt gacgctgtca
      781 gagtgtctag tcagtgcaat cagtatgaag tcttgagtct ggcttctttt cctttacaga
      841 atgcatttga gagtttccta tgtcataata tttatcactg aattgccaag tcgtaaaaca
      901 attgtagaga tgaaccctaa tccatttctt ttcccactga atgacacttg ggatgctcct
      961 agcttctgag atttgtgaga agaggcatgg taaatgttca cggagctcgg gctgggcata
     1021 aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag
     1081 ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     1141 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1201 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
     1261 ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc
     1321 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1381 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1441 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1501 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1561 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     1621 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1681 gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac
     1741 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1801 atggacgagc tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg atatcataat
     1861 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     1921 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     1981 gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc
     2041 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     2101 gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2161 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2221 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2281 cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg cggaccgagc
     2341 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     2401 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     2461 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     2521 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     2581 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     2641 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2701 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     2761 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     2821 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     2881 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     2941 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3001 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3061 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3121 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3181 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     3241 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3301 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3361 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3421 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3481 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3541 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3601 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3661 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3721 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3781 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3841 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3901 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     3961 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4021 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4081 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4141 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4201 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4261 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4321 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4381 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4441 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4501 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4561 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4621 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4681 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4741 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4801 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4861 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4921 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4981 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5041 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
