LOCUS       sequence-482722-        4999 bp    DNA     circular SYN 08-AUG-2025
DEFINITION  Transient expression of 15xGly-uFMR1-3xHA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4999)
  AUTHORS   Yang J, Xu Y, Ziehr DR, Taylor MS, Valenstein ML, Frenkel EM, Bush 
            JR, Rutter K, Stevanovski I, Shi CY, Kesavan M, Pinto RM, Deveson I,
            Bartel DP, Sabatini DM, Chivukula RR
  TITLE     Polyglycine-mediated aggregation of FAM98B disrupts tRNA processing 
            in GGC repeat disorders.
  JOURNAL   Science. 2025 Jul 17;389(6757):eado2403. doi: 
            10.1126/science.ado2403. Epub 2025 Jul 17.
  PUBMED    40674500
REFERENCE   2  (bases 1 to 4999)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science."; 
            date: "2025-07-17"; volume: "389(6757):eado2403. doi"; pages: " 
            10.1126/science.ado2403. Epub 2025 Jul 17"
COMMENT     Sequence Label: pRK5R 15xGly-uFMR1-3xHA
FEATURES             Location/Qualifiers
     source          1..4999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(117..133)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(117..133)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(130..152)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    141..157
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(165..171,172..189,190..195))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    210..231
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(348..365)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(519..1107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(599..618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1278..2069,2070..2138))
                     /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     1901..1920
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2139..2243)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2311..2329
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2367..2389)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2489..2508
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2525..2980)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2662..2683)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2874..2893
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3178..3200
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3192..3209
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3193..3209
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3229..3608
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3609..3812
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3763..3783
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3809..3833
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        4043..4061
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4043..4060
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     4074..4093
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      4160..4169
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4388..4414
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4427..4453
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4466..4492
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      4513..4522
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4526..4660
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4563..4582)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4617..4636
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        join(4729..4999,1..87)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4729..4920
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4752..4772)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(4937..4999,1..73)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
       61 ttttttggag gcctaggctt ttgcaaaaag ctgttaacag cttggctcgt aatcatgtca
      121 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
      181 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
      241 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
      301 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
      361 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
      421 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
      481 aaggccagga accgtaaaaa ggccgcgttg cttgcgtttt tccataggct ccgcccccct
      541 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      601 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
      661 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
      721 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
      781 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      841 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
      901 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
      961 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1021 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1081 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1141 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     1201 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     1261 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     1321 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     1381 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     1441 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     1501 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     1561 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     1621 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     1681 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     1741 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     1801 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     1861 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     1921 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     1981 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     2041 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     2101 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     2161 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     2221 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     2281 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     2341 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     2401 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     2461 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     2521 cataaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     2581 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagccc
     2641 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     2701 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     2761 cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     2821 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     2881 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     2941 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgta
     3001 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     3061 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     3121 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     3181 agtcacgacg ttgtaaaacg acggccagta attaattcga gctcgcccga cattgattat
     3241 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     3301 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     3361 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     3421 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     3481 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     3541 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     3601 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     3661 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     3721 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     3781 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga
     3841 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgcg
     3901 gccgggaacg gtgcattgga acgcggattc cccgtgccaa gagtgacgta agtaccgcct
     3961 atagagtcta taggcccacc cccttggctt cgttagaacg cggctacaat taatacataa
     4021 ccttatgtat catacacata cgatttaggt gacactatag aataacatcc actttgcctt
     4081 tctctccaca ggtgtccact cccaggtcca actgcacctc ggttctagct agcgctaccg
     4141 gtttaattaa ttcgaattcg ccaccatgga ggcgccgctg ccagggggcg tgcggcagcg
     4201 cggcggcggc ggcggcggcg gcggcggcgg cggcggcggc ggaggctggg cctcgagcgc
     4261 ccgcagccca cctctcgggg gcgggctccc ggcgctagca gggctgaaga gaagatggag
     4321 gagctggtgg tggaagtgcg gggctccaat ggcgctttct acaaggcacc tagcggccgc
     4381 tagcggctat ccctatgacg tgcctgatta cgccggcaca ggatcatacc cctatgatgt
     4441 gcctgactac gctggcagcg ccggataccc ttatgatgtg cctgattatg cttagtgagg
     4501 atccccagct tggccgccat ggcccaactt gtttattgca gcttataatg gttacaaata
     4561 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4621 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gggaattaat tcggcgcagc
     4681 accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa
     4741 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4801 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4861 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4921 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4981 tggctgacta atttttttt
//
