LOCUS       sequence-445043-        7699 bp    DNA     circular SYN 15-APR-2025
DEFINITION  MS2 mRNA reporter with intact FMR1 5' UTR and exon 1 sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7699)
  AUTHORS   Sirois CL, Guo Y, Li M, Wolkoff NE, Korabelnikov T, Sandoval S, Lee 
            J, Shen M, Contractor A, Sousa AMM, Bhattacharyya A, Zhao X
  TITLE     CGG repeats in the human FMR1 gene regulate mRNA localization and 
            cellular stress in developing neurons.
  JOURNAL   Cell Rep. 2024 Jun 11;43(6):114330. doi: 
            10.1016/j.celrep.2024.114330.
  PUBMED    38865241
REFERENCE   2  (bases 1 to 7699)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.";
            date: "2024-06-11"; volume: "43(6):114330. doi"; pages: " 
            10.1016/j.celrep.2024.114330"
COMMENT     Sequence Label: p40651-EFSNS-FMR1_e1-24xms2
FEATURES             Location/Qualifiers
     source          1..7699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..307
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_RNA        619..637
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        658..676
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        728..746
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        767..785
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        837..855
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        876..894
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        946..964
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        985..1003
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1055..1073
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1094..1112
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1164..1182
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1203..1221
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1273..1291
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1312..1330
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1382..1400
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1421..1439
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1491..1509
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1530..1548
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1600..1618
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1639..1657
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1709..1727
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1748..1766
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1818..1836
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1857..1875
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_feature    1924..2512
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1977..1997)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2395..2406)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2587..2820
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(2939..2957)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3025..3129
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3130..3198,3199..3990)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3348..3367)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4161..4749
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4650..4669
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4903..4920
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5160..5181
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5196..5201,5202..5219,5220..5226)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5234..5250
                     /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     5239..5261
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5258..5274
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5258..5274
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5324..5343
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             5535..6168
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    6215..6340
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6837..7070
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7255..7299
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    join(7592..7699,1..10)
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggacta gttaggtctt
       61 gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
      121 ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg
      181 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
      241 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
      301 aacacagggt ttcggtttca cttccggtgg agggccgcct ctgagcgggc ggcgggccga
      361 cggcgagcgc gggcggcggc ggtgacggag gcgccgctgc cagggggcgt gcggcagcgc
      421 gcggcggcgg cnggcggagg cggcggcggc ggcggcggcg gcggcggcgg ctgggcctcg
      481 agcgcccgca gcccacctct cgggggcggg ctcccggcgc tagcagggct gaagagaaga
      541 tggaggagct ggtggtggaa gtgcggggct ccaatggcgc tttctacaag gatcctaagg
      601 tacctaattg cctagaaaac atgaggatca cccatgtctg caggtcgact ctagaaaaca
      661 tgaggatcac ccatgtctgc agtattcccg ggttcattag atcctaaggt acctaattgc
      721 ctagaaaaca tgaggatcac ccatgtctgc aggtcgactc tagaaaacat gaggatcacc
      781 catgtctgca gtattcccgg gttcattaga tcctaaggta cctaattgcc tagaaaacat
      841 gaggatcacc catgtctgca ggtcgactcy agaaaacatg aggatcaccc atgtctgcag
      901 tattcccggg ttcattagat cctaaggtac ctaattgcct agaaaacatg aggatcaccc
      961 atgtctgcag gtcgactcta gaaaacatga ggatcaccca tgtctgcagt attcccgggt
     1021 tcattagatc ctaaggtacc taattgccta gaaaacatga ggatcaccca tgtctgcagg
     1081 tcgactctag aaaacatgag gatcacccat gtctgcagta ttcccgggtt cattagatcc
     1141 taaggtacct aattgcctag aaaacatgag gatcacccat gtctgcaggt cgactcyaga
     1201 aaacatgagg atcacccatg tctgcagtat tcccgggttc attagatcct aaggtaccta
     1261 attgcctaga aaacatgagg atcacccatg tctgcaggtc gactctagaa aacatgagga
     1321 tcacccatgt ctgcagtatt cccgggttca ttagatccta aggtacctaa ttgcctagaa
     1381 aacatgagga tcacccatgt ctgcaggtcg actctagaaa acatgaggat cacccatgtc
     1441 tgcagtattc ccgggttcat tagatcctaa ggtacctaat tgcctagaaa acatgaggat
     1501 cacccatgtc tgcaggtcga ctctagaaaa catgaggatc acccatgtct gcagtattcc
     1561 cgggttcatt agatcctaag gtacctaatt gcctagaaaa catgaggatc acccatgtct
     1621 gcaggtcgac tctagaaaac atgaggatca cccatgtctg cagtattccc gggttcatta
     1681 gatcctaagg tacctaattg cctagaaaac atgaggatca cccatgtctg caggtcgact
     1741 ctagaaaaca tgaggatcac ccatgtctgc agtattcccg ggttcattag atcctaaggt
     1801 acctaattgc ctagaaaaca tgaggatcac ccatgtctgc aggtcgactc cagaaaacat
     1861 gaggatcacc catgtctgca gtattcccgg gttcattaga tctgcgcgcg atcgatagat
     1921 cctaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     1981 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2041 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     2101 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     2161 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     2221 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     2281 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     2341 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     2401 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     2461 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctgagatc
     2521 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     2581 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     2641 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     2701 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     2761 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     2821 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     2881 agagtgagag gcccgggtta attaaggaaa gggctagatc attcttgaag acgaaagggc
     2941 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     3001 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3061 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3121 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3181 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3241 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3301 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3361 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3421 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3481 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3541 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3601 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3661 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3721 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3781 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3841 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3901 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3961 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4021 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4081 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4141 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4201 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4261 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4321 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4381 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4441 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4501 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4561 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4621 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4681 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4741 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4801 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4861 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4921 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     4981 tgcagcaagc tcatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     5041 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5101 agctccccgt ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     5161 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     5221 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgacatgatt
     5281 acgaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5341 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
     5401 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
     5461 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
     5521 acaaacttag tagttggaag ggctaattca ctcccaaaga agacaagata tccttgatct
     5581 gtggatctac cacacacaag gctacttccc tgattagcag aactacacac cagggccagg
     5641 ggtcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg agccagataa
     5701 ggtagaagag gccaataaag gagagaacac cagcttgtta caccctgtga gcctgcatgg
     5761 gatggatgac ccggagagag aagtgttaga gtggaggttt gacagccgcc tagcatttca
     5821 tcacgtggcc cgagagctgc atccggagta cttcaagaac tgctgatatc gagcttgcta
     5881 caagggactt tccgctgggg actttccagg gaggcgtggc ctgggcggga ctggggagtg
     5941 gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg tctctctggt
     6001 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     6061 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     6121 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
     6181 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
     6241 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
     6301 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
     6361 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
     6421 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
     6481 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
     6541 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
     6601 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
     6661 gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg agatatgagg
     6721 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     6781 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     6841 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     6901 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     6961 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     7021 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     7081 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     7141 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     7201 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     7261 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     7321 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     7381 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     7441 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
     7501 ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg acggtatcgc
     7561 cgaattcaca aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt
     7621 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
     7681 aaaaacaaat tacaaaaat
//
