LOCUS       sequence-441703-        7909 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express EGFP-fusion protein of FXR1 isoform a (CC1-CC1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7909)
  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 7909)
  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-FXR1a-CC1-CC1
FEATURES             Location/Qualifiers
     source          1..7909
                     /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(1135..1137,1138..1140,1141..1851)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1135..1137,1138..1140,1141..1851)
                     /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(1441..1460)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1788..1809
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3752..3769)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3758..3982
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4028..4456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4115..4134)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4325..4346
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4465..4485)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4470..4799
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4650..4785
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4712..4731
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4812..5411
                     /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(4812..4831)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5308..5328
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5585..5706
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5622..5641)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5676..5695
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5755..5771)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5755..5771)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5768..5790)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5779..5795
                     /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(5803..5809,5810..5827,5828..5833))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5848..5869
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5986..6003)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6157..6742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6237..6256)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6913..7704,7705..7773))
                     /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     7536..7555
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7774..7878)
                     /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 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      961 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1021 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1081 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatggtg
     1141 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1201 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1261 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1321 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     1381 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1441 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1501 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1561 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1621 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1681 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1741 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1801 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa ggcggagctg
     1861 acggtggagg ttcgcggctc taacggggct ttctacaagg gatttatcaa agatgttcat
     1921 gaagactccc ttacagttgt ttttgaaaat aattggcaac cagaacgcca ggttccattt
     1981 aatgaagtta gattaccacc accacctgat ataaaaaaag aaattagtga aggagatgaa
     2041 gtagaggtat attcaagagc aaatgaccaa gagccatgtg ggtggtggtt ggctaaagtt
     2101 cggatgatga aaggagaatt ttatgtcatt gaatatgctg cttgtgacgc tacttacaat
     2161 gaaatagtca catttgaacg acttcggcct gtcaatcaaa ataaaactgt caaaaaaaat
     2221 accttcttta aatgcacagt ggatgttcct gaggatttga gagaggcgtg tgctaatgaa
     2281 aatgcacata aagattttaa gaaagcagta ggagcatgca gaatttttta ccatccagaa
     2341 acaacacagc taatgatact gtctgccagt gaagcaactg tgaagagagt aaacatctta
     2401 agtgacatgc atttgcgaag tattcgtacg aagttgatgc ttatgtccag aaatgaagag
     2461 gccactaagc atttagaatg cacaaaacaa cttgcagcag cttttcatga ggaatttgtt
     2521 gtgagagaag atttaatggg cctggcaata ggaacacatg gtagtaacat ccagcaagct
     2581 aggaaggttc ctggagttac cgccattgag ctagatgaag atactggaac attcagaatc
     2641 tacggagaga gtgctgatgc tgtaaaaaag gctagaggtt tcttggaatt tgtggaggat
     2701 tttattcagg ttcctaggaa tctcgttgga aaagtaattg gaaaaaatgg caaagttatt
     2761 caagaaatag tggacaaatc tggtgtggtt cgagtgagaa ttgaagggga caatgaaaat
     2821 aaattaccca gagaagacgg tatggttcca tttgtatttg ttggcactaa agaaagcatt
     2881 ggaaatgtgc aggttcttct agagtatcat attgcctatc taaaggaaaa tgaagaggcc
     2941 actaagcatt tagaatgcac aaaacaactt gcagcagctt ttggttctag gtcttatagc
     3001 ggaagaggca gaggtcgtcg gggacctaat tacacctccg gttatggtac aaattctgag
     3061 ctgtctaacc cctctgaaac ggaatctgag cgtaaagacg agctgagtga ttggtcattg
     3121 gcaggagaag atgatcgaga cagccgacat cagcgtgaca gcaggagacg cccaggagga
     3181 agaggcagaa gtgtttcagg gggtcgaggt cgtggtggac cacgtggtgg caaatcctcc
     3241 atcagttctg tgctcaaaga tccagacagc aatccataca gcttacttga taatacagaa
     3301 tcagatcaga ctgcagacac tgatgcatcc gagagccacc attccactaa ccgtcgtagg
     3361 cggtctcgta gacgaaggac tgatgaagat gctgttctga tggatggaat gactgaatct
     3421 gatacagctt cagttaatga aaatgggcta gtcacagttg cagattatat ttctagagct
     3481 gagtctcaga gcagacaaag aaacctccca agggaaactt tggctaaaaa caagaaagaa
     3541 atggcaaaag atgtgattga agagcatggt ccttcagaaa aggcaataaa cggcccaact
     3601 agtgcttctg gcgatgacat ttctaagcta cagcgtactc caggagaaga aaagattaat
     3661 accttaaaag aagaaaacac tcaagaagca gcagtcctga atggtgtttc ataactcgag
     3721 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     3781 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3841 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3901 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3961 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     4021 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4081 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4141 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4201 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4261 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4321 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4381 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4441 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4501 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4561 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4621 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4681 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4741 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4801 agctcccaac catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     4861 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     4921 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     4981 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     5041 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     5101 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     5161 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     5221 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     5281 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     5341 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     5401 ccggtgcctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5461 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5521 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5581 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5641 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5701 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     5761 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5821 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5881 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5941 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6001 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6061 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6121 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6181 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6241 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6301 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6361 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6421 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6481 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6541 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6601 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6661 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     6721 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6781 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6841 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6901 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6961 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7021 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7081 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7141 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7201 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7261 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7321 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7381 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7441 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7501 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7561 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7621 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7681 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7741 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7801 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7861 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
