LOCUS       sequence-458915-        7325 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express mEGFP-fusion protein of FMR1 fragment (1-491) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7325)
  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 7325)
  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. 5 
            August 2024. doi: 10.1016/j.cell.2024.07.015."
COMMENT     Sequence Label: pcDNA3puro-mGFP-FMR1iso1-N2(1-491)
FEATURES             Location/Qualifiers
     source          1..7325
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(905..907,908..910,911..1621)
                     /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(950..971)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1211..1230)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1558..1579
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3165..3182)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3171..3395
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3441..3869
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3528..3547)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3738..3759
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3878..3898)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3883..4212
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4063..4198
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4125..4144
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4225..4824
                     /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(4225..4244)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4721..4741
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4998..5119
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5035..5054)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5089..5108
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5168..5184)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5168..5184)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5181..5203)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5192..5208
                     /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(5216..5222,5223..5240,5241..5246))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5261..5282
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5399..5416)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5570..6158)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5650..5669)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6329..7120,7121..7189))
                     /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     6952..6971
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7190..7294)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 aaccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      961 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1021 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1081 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     1141 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1201 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1261 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1321 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1381 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1441 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1501 ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     1561 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1621 ggaggagctg gtggtggaag tgcggggctc caatggcgct ttctacaagg catttgtaaa
     1681 ggatgttcat gaagattcaa taacagttgc atttgaaaac aactggcagc ctgataggca
     1741 gattccattt catgatgtca gattcccacc tcctgtaggt tataataaag atataaatga
     1801 aagtgatgaa gttgaggtgt attccagagc aaatgaaaaa gagccttgct gttggtggtt
     1861 agctaaagtg aggatgataa agggtgagtt ttatgtgata gaatatgcag catgtgatgc
     1921 aacttacaat gaaattgtca caattgaacg tctaagatct gttaatccca acaaacctgc
     1981 cacaaaagat actttccata agatcaagct ggatgtgcca gaagacttac ggcaaatgtg
     2041 tgccaaagag gcggcacata aggattttaa aaaggcagtt ggtgcctttt ctgtaactta
     2101 tgatccagaa aattatcagc ttgtcatttt gtccatcaat gaagtcacct caaagcgagc
     2161 acatatgctg attgacatgc actttcggag tctgcgcact aagttgtctc tgataatgag
     2221 aaatgaagaa gctagtaagc agctggagag ttcaaggcag cttgcctcga gatttcatga
     2281 acagtttatc gtaagagaag atctgatggg tctagctatt ggtactcatg gtgctaatat
     2341 tcagcaagct agaaaagtac ctggggtcac tgctattgat ctagatgaag atacctgcac
     2401 atttcatatt tatggagagg atcaggatgc agtgaaaaaa gctagaagct ttctcgaatt
     2461 tgctgaagat gtaatacaag ttccaaggaa cttagtaggc aaagtaatag gaaaaaatgg
     2521 aaagctgatt caggagattg tggacaagtc aggagttgtg agggtgagga ttgaggctga
     2581 aaatgagaaa aatgttccac aagaagagga aattatgcca ccaaattccc ttccttccaa
     2641 taattcaagg gttggaccta atgccccaga agaaaaaaaa catttagata taaaggaaaa
     2701 cagcacccat ttttctcaac ctaacagtac aaaagtccag agggtgttag tggcttcatc
     2761 agttgtagca ggggaatccc agaaacctga actcaaggct tggcagggta tggtaccatt
     2821 tgtttttgtg ggaacaaagg acagcatcgc taatgccact gttcttttgg attatcacct
     2881 gaactattta aaggaagtag accagttgcg tttggagaga ttacaaattg atgagcagtt
     2941 gcgacagatt ggagctagtt ctagaccacc accaaatcgt acagataagg aaaaaagcta
     3001 tgtgactgat gatggtcaag gaatgggtcg aggtagtaga ccttacagaa atagggggca
     3061 cggcagacgc ggtcctggat atacttcagg ataagaattc tgcagatatc cagcacagtg
     3121 gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     3181 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3241 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3301 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3361 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3421 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3481 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3541 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3601 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3661 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3721 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3781 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3841 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3901 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3961 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4021 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4081 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4141 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4201 ggcttttgca aaaagctccc aaccatgacc gagtacaagc ccacggtgcg cctcgccacc
     4261 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     4321 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     4381 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     4441 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     4501 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     4561 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     4621 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     4681 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     4741 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     4801 atgacccgca agcccggtgc ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4861 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4921 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4981 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5041 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5101 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5161 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5221 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5281 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5341 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5401 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5461 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5521 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5581 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5641 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5701 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5761 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5821 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5881 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5941 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6001 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6061 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6121 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6181 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6241 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6301 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6361 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6421 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6481 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6541 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6601 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6661 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6721 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6781 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6841 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6901 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6961 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7021 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7081 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7141 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7201 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7261 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7321 acgtc
//
