LOCUS       sequence-445052-        7647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MS2 mRNA reporter with FMR1 5' UTR (CGG repeats removed) and exon 1 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7647)
  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 7647)
  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: "2024-06-11"; volume: "43(6):114330. doi"; pages: " 
            10.1016/j.celrep.2024.114330"
COMMENT     Sequence Label: p40651-EFSNS-FMR1_e1-Delta-CGG-24xms2
FEATURES             Location/Qualifiers
     source          1..7647
                     /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        567..585
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        606..624
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        676..694
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        715..733
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        785..803
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        824..842
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        894..912
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        933..951
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1003..1021
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1042..1060
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1112..1130
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1151..1169
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1221..1239
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1260..1278
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1330..1348
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1369..1387
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1439..1457
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1478..1496
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1548..1566
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1587..1605
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1657..1675
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1696..1714
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1766..1784
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1805..1823
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_feature    1872..2460
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1925..1945)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2343..2354)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2535..2768
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(2887..2905)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2973..3077
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3078..3146,3147..3938)
                     /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(3296..3315)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4109..4697
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4598..4617
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4851..4868
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5108..5129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5144..5149,5150..5167,5168..5174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5182..5198
                     /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     5187..5209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5206..5222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5206..5222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5272..5291
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             5483..6116
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    6163..6288
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6785..7018
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7203..7247
                     /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(7540..7647,1..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 tgggcctcga gcgcccgcag cccacctctc gggggcgggc tcccggcgct agcagggctg
      481 aagagaagat ggaggagctg gtggtggaag tgcggggctc caatggcgct ttctacaagg
      541 atctaaggta cctaattgcc tagaaaacat gaggatcacc catgtctgca ggtcgactct
      601 agaaaacatg aggatcaccc atgtctgcag tattcccggg ttcattagat cctaaggtac
      661 ctaattgcct agaaaacatg aggatcaccc atgtctgcag gtcgactcta gaaaacatga
      721 ggatcaccca tgtctgcagt attcccgggt tcattagatc ctaaggtacc taattgccta
      781 gaaaacatga ggatcaccca tgtctgcagg tcgactcyag aaaacatgag gatcacccat
      841 gtctgcagta ttcccgggtt cattagatcc taaggtacct aattgcctag aaaacatgag
      901 gatcacccat gtctgcaggt cgactctaga aaacatgagg atcacccatg tctgcagtat
      961 tcccgggttc attagatcct aaggtaccta attgcctaga aaacatgagg atcacccatg
     1021 tctgcaggtc gactctagaa aacatgagga tcacccatgt ctgcagtatt cccgggttca
     1081 ttagatccta aggtacctaa ttgcctagaa aacatgagga tcacccatgt ctgcaggtcg
     1141 actcyagaaa acatgaggat cacccatgtc tgcagtattc ccgggttcat tagatcctaa
     1201 ggtacctaat tgcctagaaa acatgaggat cacccatgtc tgcaggtcga ctctagaaaa
     1261 catgaggatc acccatgtct gcagtattcc cgggttcatt agatcctaag gtacctaatt
     1321 gcctagaaaa catgaggatc acccatgtct gcaggtcgac tctagaaaac atgaggatca
     1381 cccatgtctg cagtattccc gggttcatta gatcctaagg tacctaattg cctagaaaac
     1441 atgaggatca cccatgtctg caggtcgact ctagaaaaca tgaggatcac ccatgtctgc
     1501 agtattcccg ggttcattag atcctaaggt acctaattgc ctagaaaaca tgaggatcac
     1561 ccatgtctgc aggtcgactc tagaaaacat gaggatcacc catgtctgca gtattcccgg
     1621 gttcattaga tcctaaggta cctaattgcc tagaaaacat gaggatcacc catgtctgca
     1681 ggtcgactct agaaaacatg aggatcaccc atgtctgcag tattcccggg ttcattagat
     1741 cctaaggtac ctaattgcct agaaaacatg aggatcaccc atgtctgcag gtcgactcca
     1801 gaaaacatga ggatcaccca tgtctgcagt attcccgggt tcattagatc tgcgcgcgat
     1861 cgatagatcc taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     1921 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     1981 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     2041 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     2101 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     2161 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     2221 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     2281 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     2341 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     2401 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     2461 ctgagatcct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     2521 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
     2581 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     2641 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     2701 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     2761 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     2821 aatatcagag agtgagaggc ccgggttaat taaggaaagg gctagatcat tcttgaagac
     2881 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     2941 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3001 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3061 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3121 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3181 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3241 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3301 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     3361 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3421 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3481 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3541 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3601 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3661 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3721 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3781 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3841 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3901 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3961 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4021 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4081 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4141 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4201 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4261 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4321 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4381 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4441 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4501 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4561 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4621 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4681 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4741 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4801 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4861 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     4921 ttcattaatg cagcaagctc atggctgact aatttttttt atttatgcag aggccgaggc
     4981 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     5041 ttgcaaaaag ctccccgtgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     5101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     5161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     5221 acatgattac gaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5281 ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa
     5341 atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt
     5401 tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa
     5461 tatgtactac aaacttagta gttggaaggg ctaattcact cccaaagaag acaagatatc
     5521 cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca
     5581 gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
     5641 ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc
     5701 ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta
     5761 gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga
     5821 gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact
     5881 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc
     5941 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     6001 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     6061 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
     6121 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
     6181 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     6241 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     6301 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     6361 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
     6421 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     6481 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     6541 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     6601 aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat cttcagacct ggaggaggag
     6661 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     6721 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     6781 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     6841 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     6901 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     6961 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     7021 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     7081 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     7141 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     7201 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     7261 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     7321 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     7381 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     7441 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac
     7501 ggtatcgccg aattcacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     7561 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     7621 agaattacaa aaacaaatta caaaaat
//
