LOCUS       sequence-176829-        6592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse FMRP-EGFP fusion with phosphorylation mimic S499D..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6592)
  AUTHORS   Stetler A, Winograd C, Sayegh J, Cheever A, Patton E, Zhang X, 
            Clarke S, Ceman S
  TITLE     Identification and characterization of the methyl arginines in the 
            fragile X mental retardation protein Fmrp.
  JOURNAL   Hum Mol Genet. 2006 Jan 1;15(1):87-96. Epub 2005 Nov 30.
  PUBMED    16319129
REFERENCE   2  (bases 1 to 6592)
  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: "Hum Mol 
            Genet."; date: "2006-01-1"; volume: "15(1)"; pages: "87-96. Epub 
            2005 Nov 30"
COMMENT     Sequence Label: p-EGFP-C1-Flag-mFmr1(D)
FEATURES             Location/Qualifiers
     source          1..6592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1312..1335
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3320..3441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3357..3376)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3411..3430
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3448..3903)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3585..3606)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3797..3816
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3930..4034
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4036..4393
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4036..4227
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4059..4079)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4244..4379
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4428..5222
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4428..5222
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4482..4501)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5092..5111
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5410..5429)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5454..5501
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5830..6418
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6319..6338
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atctcgagct caagcttcga attcgatgga ggactacaag
     1321 gacgacgatg acaaggagct ggtggtggaa gtgcggggct ccaatggcgc tttctacaag
     1381 gcatttgtaa aagatgtcca tgaagattct ataacagttg cttttgaaaa caactggcaa
     1441 ccagagagac agattccatt ccatgatgtg agattcccac cacctgtagg ttataataaa
     1501 gatataaacg aaagtgatga agttgaggtt tattccagag caaatgaaaa agagccttgc
     1561 tgttggtggt tagctaaagt gaggatgata aagggtgagt tttatgtgat agaatatgca
     1621 gcatgtgatg ctacgtataa tgaaattgtc acaattgagc gtctacgatc tgttaatccc
     1681 aacaaacctg ctacaaaaga tactttccat aagatcaagc tggaggtgcc agaagattta
     1741 cgacaaatgt gtgccaaaga atcagcacat aaggatttta aaaaggcagt tggtgccttc
     1801 tctgtaactt atgatccaga aaattatcag ctggtaattt tgtccatcaa tgaagtcacc
     1861 tcaaagcgag cccacatgtt gattgacatg cactttcgaa gtctgcgcac caagttgtct
     1921 cttatactga gaaatgaaga agccagtaaa caactggaga gttcaaggca gcttgcctca
     1981 agatttcatg aacagtttat cgtacgagaa gatctgatgg gtttagctat tggtactcat
     2041 ggtgctaata ttcagcaagc tagaaaagtg cctggcgtca ctgctattga tttagatgag
     2101 gatacctgca catttcatat ttatggagag gatcaagatg cagtgaaaaa ggctagaagc
     2161 tttctggaat ttgctgaaga tgtcatacag gttccacgaa acttagtagg caaagtaata
     2221 ggaaaaaatg gaaagctgat tcaagagatc gtggacaagt caggagttgt gagggtgagg
     2281 attgaggctg aaaatgagaa aagtgtccca caagaagagg aaattatgcc accaagttcc
     2341 ctaccttcca ataattcaag ggttggacct aactcctctg aagaaaagaa acatttagat
     2401 acaaaggaaa acacccattt ttctcaacct aacagtacaa aagtccagag ggtgttagtg
     2461 gtttcatcaa ttgtagcagg gggaccccag aaacctgaac ccaaggcttg gcagggtatg
     2521 gtaccatttg tttttgtggg aacaaaagac agcatcgcta atgccactgt tcttttggat
     2581 tatcacctga actatttaaa ggaagtagac cagttgcgtt tggagagatt acaaattgat
     2641 gagcagttgc gacaaattgg agctagttct agaccaccac caaatcgtac agataaggaa
     2701 aaaggctatg tgactgatga tggtcaagga atgggtcgag gtagtagacc ttacagaaat
     2761 agggggcacg gcagacgcgg tcctggatat acttcaggaa ctaattctga agcatcaaat
     2821 gctgacgaaa cagaatctga ccacagagac gaactcagtg attggtcatt agctccaaca
     2881 gaggaagaga gggagagctt cctgcgcaga ggagacggac ggcggcgtgg aggaggagga
     2941 agaggacaag gaggaagagg aagaggagga ggcttcaaag gaaacgacga tcattcccga
     3001 acagataatc gtccacgtaa tccaagagag gctaaaggaa gaacagctga tggatccctg
     3061 cagagtgcct ccagtgaagg gagccggctg cgcacgggta aagatcgtaa ccagaagaag
     3121 gaaaagccag acagcgtaga tgggctgcaa ccgctggtga atggagtacc ctaaataagc
     3181 cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata ccacatttgt
     3241 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3301 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3361 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3421 caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     3481 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3541 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3601 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3661 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     3721 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     3781 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     3841 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     3901 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3961 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4021 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     4081 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4141 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4201 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4261 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4321 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4381 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4441 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4501 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4561 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     4621 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4681 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4741 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4801 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4861 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4921 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     4981 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5041 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5101 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5161 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5221 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5281 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5341 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     5401 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     5461 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     5521 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5581 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5641 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     5701 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5761 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5821 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5881 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5941 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6001 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6061 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6121 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6181 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6241 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6301 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6361 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6421 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6481 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
     6541 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cg
//
