LOCUS       sequence-411197-        5115 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of FMRP RNA for gene silencing in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5115)
  AUTHORS   Ferron L, Nieto-Rostro M, Cassidy JS, Dolphin AC
  TITLE     Fragile X mental retardation protein controls synaptic vesicle 
            exocytosis by modulating N-type calcium channel density.
  JOURNAL   Nat Commun. 2014 Apr 7;5:3628. doi: 10.1038/ncomms4628.
  PUBMED    24709664
REFERENCE   2  (bases 1 to 5115)
  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: "Nat 
            Commun."; date: "2014-04-7"; volume: "5:3628. doi"; pages: " 
            10.1038/ncomms4628"
COMMENT     Sequence Label: FMRP shRNA
FEATURES             Location/Qualifiers
     source          1..5115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..77
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             139..942
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(202..221)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     812..831
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    956..1417
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     promoter        complement(1483..1795)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     complement(1544..1564)
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     complement(1804..1820)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1805..1821)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1845..1864)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1846..1864)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1871..1888)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1871..1887)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1880..1902)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2029..2484)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2166..2187)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2378..2397
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2511..2615
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2616..2684,2685..3476)
                     /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(2834..2853)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3647..4235
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4136..4155
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4389..4406
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4523..4544
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4559..4564,4565..4582,4583..4589)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4597..4613
                     /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)."
     primer_bind     4602..4624
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4621..4637
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4621..4637
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4658..4676
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        join(4741..5115,1..125)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4776..4799)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat
       61 tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc
      121 tttcgacctg cagccaatat gggatcggcc attgaacaag atggattgca cgcaggttct
      181 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
      241 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
      301 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
      361 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
      421 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
      481 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
      541 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
      601 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
      661 gccaggctca aggcgcgcat gcccgacggc gatgatctcg tcgtgaccca tggcgatgcc
      721 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
      781 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
      841 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
      901 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gaggggatcg atccgctgta
      961 agtctgcaga aattgatgat ctattaaaca ataaagatgt ccactaaaat ggaagttttt
     1021 cctgtcatac tttgttaaga agggtgagaa cagagtacct acattttgaa tggaaggatt
     1081 ggagctacgg gggtgggggt ggggtgggat tagataaatg cctgctcttt actgaaggct
     1141 ctttactatt gctttatgat aatgtttcat agttggatat cataatttaa acaagcaaaa
     1201 ccaaattaag ggccagctca ttcctcccac tcatgatcta tagatctata gatctctcgt
     1261 gggatcattg tttttctctt gattcccact ttgtggttct aagtactgtg gtttccaaat
     1321 gtgtcagttt catagcctga agaacgagat cagcagcctc tgttccacat acacttcatt
     1381 ctcagtattg ttttgccaag ttctaattcc atcagaagct gactctagaa aaagtgatga
     1441 agttgaggtt tatctcttga ataaacctca acttcatcac caaacaaggc ttttctccaa
     1501 gggatattta tagtctcaaa acacacaatt actttacagt tagggtgagt ttccttttgt
     1561 gctgtttttt aaaataataa tttagtattt gtatctctta tagaaatcca agcctatcat
     1621 gtaaaatgta gctagtatta aaaagaacag attatctgtc ttttatcgca cattaagcct
     1681 ctatagttac taggaaatat tatatgcaaa ttaaccgggg caggggagta gccgagcttc
     1741 tcccacaagt ctgtgcgagg gggccggcgc gggcctagag atggcggcgt cggataagct
     1801 tatcgatacc gtcgacctcg agggggggcc cggtacccaa ttcgccctat agtgagtcgt
     1861 attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     1921 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     1981 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcaa attgtaagcg
     2041 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2101 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2161 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2221 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2281 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2341 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2401 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2461 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2521 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2581 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     2641 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2701 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     2761 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     2821 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     2881 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     2941 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3001 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3061 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3121 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3181 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3241 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3301 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3361 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3421 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3481 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3541 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3601 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3661 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3721 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3781 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3841 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3901 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3961 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4021 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4081 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4141 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4201 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4261 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4321 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4381 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     4441 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     4501 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     4561 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     4621 caggaaacag ctatgaccat gattacgcca agctcgaaat taaccctcac taaagggaac
     4681 aaaagctgga gctccaccgc ggtggcggcc gctcgagggc ccctgcaggt caattctacc
     4741 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
     4801 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
     4861 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
     4921 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
     4981 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
     5041 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg
     5101 gtgggtccgg gggcg
//
