LOCUS       sequence-445738-        5311 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 5311)
  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 5311)
  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: HCT131_rAAV-eHGT_1498m-minBglobin-SYFP2-WPRE3-...
FEATURES             Location/Qualifiers
     source          1..5311
                     /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      1323..1332
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1329..1331,1332..1334,1335..2045)
                     /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(1374..1395)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1635..1654)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1982..2003
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2138..2158)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2345..2456
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2574..2714
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2574..2703
                     /label=AAV2 ITR
     rep_origin      2789..3244
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2876..2895)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3086..3107
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3261..3280)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3380..3402
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3440..3458)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3526..3630
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3631..3699,3700..4491)
                     /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(3849..3868)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4662..5250
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5151..5170
                     /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 gcgtactcat aaatgcctac gacttctttc ttccagcctg
      181 ggctctcaca ctttactgtc ccatctctca actttattaa ttatcttaca taatatccag
      241 aagcagtcaa acgcctgcag aaagcttcga gtagtaagaa agtcctcctc tatcttcctg
      301 gcctatggct atgctaagtg gtgacctaac agggtcggcc tgcctgccaa cagccagccc
      361 aggtctgagt ttactattcc cagaccttcc tactgtatga aacccctttc tcacactttc
      421 caattactct gcctctagtc ttttcaatcc atcccctaca tatcgaccag gtcagtctcc
      481 tttgtgctcc tattcaaatt attctgttca aagattgtca gcagttctat attgtacaca
      541 tcaggaaaat ctaagcttct cagtctaatt ttcaacgatg ttgaaagagg tggtacatcg
      601 catgcctaga agatgcttgc aggtgctcac taaggatgct gttatggttc aaatgagaac
      661 tgcctcccac gggctcttag atatgaactc ttgtcctcag ttggggatgc tgtttaagga
      721 ggctgtggaa agcttagggc acggaatctt gctggagaaa gtacatctct gggagtgttt
      781 ttttttcaga atttatagtt tcttggcatt tgcagtttgt tctctcttag cttccaatgt
      841 gctgatgaag atgtgttctc ttcttcctgt ccttgttgct gttcttttct tgccattaag
      901 aacatctaga tccctgagac tgtaagcctg agcccgaata ggctcttctg aagttgactt
      961 tgttcctgct aattcatcag agtatcagaa aattcagtaa cacagatgcc aaatgaaaag
     1021 cttaaggttc cactgtccct gagtcatttg tacatcttag ccacataggc agctaagtgt
     1081 gtcacggagg tttcactctt gtgcagagcc tctagcctga attcctctct gacaacacag
     1141 cccagaatat gccattgtca ttagtttcca tgtctttgcg tctttggaac aagaggacct
     1201 tgagacaaat gagccagtag aaccatgctg agctcgggct gggcataaaa gtcagggcag
     1261 agccatctat tgcttacatt tgcttctggg atccagatct ttcgaagcta gcgctaccgg
     1321 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1381 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1441 ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct
     1501 ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc
     1561 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1621 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1681 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1741 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc
     1801 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
     1861 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1921 acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     1981 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     2041 acaagtaagt cgacggcgcg ccgcggccgc gaattcgata tcataatcaa cctctggatt
     2101 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2161 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2221 cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg
     2281 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct cgagagatct
     2341 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2401 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2461 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2521 gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc
     2581 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     2641 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     2701 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2761 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     2821 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     2881 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2941 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3001 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3061 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3121 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     3181 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     3241 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     3301 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3361 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3421 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3481 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3541 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3601 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3661 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3721 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3781 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3841 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3901 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3961 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4021 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4081 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4141 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4201 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4261 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4321 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4381 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4441 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4501 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4561 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4621 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4681 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4741 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4801 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4861 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4921 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4981 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5041 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5101 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5161 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5221 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5281 ctggcctttt gctggccttt tgctcacatg t
//
