LOCUS       sequence-448625-        7777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-fusion protein of FMR1 isoform 1 (I304N).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7777)
  AUTHORS   Chen X, Fansler MM, Janjos U, Ule J, Mayr C
  TITLE     The FXR1 network acts as signaling scaffold for actomyosin 
            remodeling.
  JOURNAL   Cell. 5 August 2024. doi: 10.1016/j.cell.2024.07.015.
  PUBMED    37961296
REFERENCE   2  (bases 1 to 7777)
  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. 5 
            August 2024. doi: 10.1016/j.cell.2024.07.015."
COMMENT     Sequence Label: pcDNA3puro-mGFP-FMR1iso1-I304N
FEATURES             Location/Qualifiers
     source          1..7777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1680..1697)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1686..1910
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1956..2384
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2043..2062)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2253..2274
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2393..2413)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2398..2727
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2578..2713
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2640..2659
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2740..3339
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2740..2759)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3236..3256
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3513..3634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3550..3569)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3604..3623
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3683..3699)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3683..3699)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3696..3718)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3707..3723
                     /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(3731..3737,3738..3755,3756..3761))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3776..3797
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3914..3931)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4085..4673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4165..4184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4844..5635,5636..5704))
                     /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     5467..5486
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5705..5809)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5884..5903)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6075..6454
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6455..6658
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6609..6629
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6703..6722
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6703..6721
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(6745..6747,6748..6750,6751..7461)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6790..6811)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7051..7070)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7398..7419
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 caacttacaa tgaaattgtc acaattgaac gtctaagatc tgttaatccc aacaaacctg
       61 ccacaaaaga tactttccat aagatcaagc tggatgtgcc agaagactta cggcaaatgt
      121 gtgccaaaga ggcggcacat aaggatttta aaaaggcagt tggtgccttt tctgtaactt
      181 atgatccaga aaattatcag cttgtcattt tgtccatcaa tgaagtcacc tcaaagcgag
      241 cacatatgct gattgacatg cactttcgga gtctgcgcac taagttgtct ctgataatga
      301 gaaatgaaga agctagtaag cagctggaga gttcaaggca gcttgcctcg agatttcatg
      361 aacagtttat cgtaagagaa gatctgatgg gtctagctat tggtactcat ggtgctaata
      421 ttcagcaagc tagaaaagta cctggggtca ctgctattga tctagatgaa gatacctgca
      481 catttcatat ttatggagag gatcaggatg cagtgaaaaa agctagaagc tttctcgaat
      541 ttgctgaaga tgtaatacaa gttccaagga acttagtagg caaagtaata ggaaaaaatg
      601 gaaagctgaa tcaggagatt gtggacaagt caggagttgt gagggtgagg attgaggctg
      661 aaaatgagaa aaatgttcca caagaagagg aaattatgcc accaaattcc cttccttcca
      721 ataattcaag ggttggacct aatgccccag aagaaaaaaa acatttagat ataaaggaaa
      781 acagcaccca tttttctcaa cctaacagta caaaagtcca gagggtgtta gtggcttcat
      841 cagttgtagc aggggaatcc cagaaacctg aactcaaggc ttggcagggt atggtaccat
      901 ttgtttttgt gggaacaaag gacagcatcg ctaatgccac tgttcttttg gattatcacc
      961 tgaactattt aaaggaagta gaccagttga gactcgagcg gctacagatt gatgagcagt
     1021 tgcgacagat tggagctagt tctagaccac caccaaatcg tacagataag gaaaaaagct
     1081 atgtgactga tgatggtcaa ggaatgggtc gaggtagtag accttacaga aatagggggc
     1141 acggcagacg cggtcctgga tatacttcag gaactaattc tgaagcatca aatgcttctg
     1201 aaacagaatc tgaccacaga gacgaactca gtgattggtc attagctcca acagaggaag
     1261 agagggagag cttcctgcgc agaggagacg gacggcggcg tggaggggga ggaagaggac
     1321 aaggaggaag aggacgtgga ggaggcttca aaggaaacga cgatcactcc cgaacagata
     1381 atcgtccacg taatccaaga gaggctaaag gaagaacaac agatggatcc cttcagatca
     1441 gagttgactg caataatgaa aggagtgtcc acactaaaac attacagaat acctccagtg
     1501 aaggtagtcg gctgcgcacg ggtaaagatc gtaaccagaa gaaagagaag ccagacagcg
     1561 tggatggtca gcaaccactc gtgaatggag taccctaaac gaattcgcgg ccgcactcga
     1621 gattccagca cagtggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc
     1681 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     1741 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     1801 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     1861 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     1921 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     1981 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2041 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2101 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2161 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2221 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2281 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     2341 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     2401 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2461 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     2521 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     2581 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2641 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     2701 ttttttggag gcctaggctt ttgcaaaaag ctcccaacca tgaccgagta caagcccacg
     2761 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     2821 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     2881 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     2941 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     3001 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     3061 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     3121 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     3181 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     3241 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     3301 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag cgggactctg gggttcgaaa
     3361 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     3421 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     3481 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     3541 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3601 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     3661 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3721 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3781 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3841 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3901 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3961 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4021 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4081 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4141 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4201 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4261 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4321 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4381 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4441 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4501 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4561 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4621 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4681 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4741 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4801 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4861 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4921 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4981 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5041 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5101 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5161 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5221 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5281 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5341 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5401 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5461 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5521 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5581 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5641 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5701 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5761 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5821 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     5881 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     5941 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     6001 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     6061 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     6121 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     6181 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     6241 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     6301 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     6361 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     6421 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     6481 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcga
     6541 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     6601 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
     6661 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag
     6721 ggagacccaa gctggctagc aaccatggtg agcaagggcg aggagctgtt caccggggtg
     6781 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     6841 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     6901 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     6961 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     7021 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     7081 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     7141 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     7201 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     7261 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     7321 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc
     7381 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     7441 ggcatggacg agctgtacaa aaaagcaggc tccaccatgg aggagctggt ggtggaagtg
     7501 cggggctcca atggcgcttt ctacaaggca tttgtaaagg atgttcatga agattcaata
     7561 acagttgcat ttgaaaacaa ctggcagcct gataggcaga ttccatttca tgatgtcaga
     7621 ttcccacctc ctgtaggtta taataaagat ataaatgaaa gtgatgaagt tgaggtgtat
     7681 tccagagcaa atgaaaaaga gccttgctgt tggtggttag ctaaagtgag gatgataaag
     7741 ggtgagtttt atgtgataga atatgcagca tgtgatg
//
