LOCUS       sequence-448470-        7682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-fusion protein of FXR1 isoform a (V361P).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7682)
  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 7682)
  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-CC2mut (V361P)
FEATURES             Location/Qualifiers
     source          1..7682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     470..489
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(502..831)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      516..651
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(570..589)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     816..836
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(845..1273)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(955..976)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1167..1186
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    1319..1543
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1532..1549
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(3450..4160,4161..4163,4164..4166))
                     /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(3492..3513)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3841..3860
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4100..4121
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4189..4208)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4190..4208)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4253..4456)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4282..4302)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4457..4836
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5008..5027
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5102..5206
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5207..5275,5276..6067)
                     /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     complement(5425..5444)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6238..6826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6727..6746
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6980..6997
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7114..7135
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7150..7155,7156..7173,7174..7180)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7188..7204
                     /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     7193..7215
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7212..7228
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7212..7228
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    7277..7398
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7288..7307)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7342..7361
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(7572..7682,1..489))
                     /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(7655..7675)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc gcctccactc cggggagcac
       61 gacggcgctg cccagaccct tgccctggtg gtcgggcgag acgccgacgg tggccaggaa
      121 ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct tccatctgtt gctgcgcggc
      181 cagccgggaa ccgctcaact cggccatgcg cgggccgatc tcggcgaaca ccgcccccgc
      241 ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg ccgtcgtccg cgacccacac
      301 cttgccgatg tcgagcccga cgcgcgtgag gaagagttct tgcagctcgg tgacccgctc
      361 gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg tagtcggcga acgcggcggc
      421 gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg aggcgcaccg tgggcttgta
      481 ctcggtcatg gttgggagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
      541 cttctggaat agctcagagg cagaggcggc ctcggcctct gcataaataa aaaaaattag
      601 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
      661 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
      721 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
      781 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
      841 aattaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
      901 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
      961 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     1021 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     1081 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     1141 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     1201 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     1261 ctacagggcg cgtggggata ccccctagag ccccagctgg ttctttccgc ctcagaagcc
     1321 atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc ccccttgctg
     1381 tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt
     1441 tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg
     1501 ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc tgatcagcgg
     1561 gtttaaacgg gccctctaga ctcgagttat gaaacaccat tcaggactgc tgcttcttga
     1621 gtgttttctt cttttaaggt attaatcttt tcttctcctg gagtacgctg tagcttagaa
     1681 atgtcatcgc cagaagcact agttgggccg tttattgcct tttctgaagg accatgctct
     1741 tcaatcacat cttttgccat ttctttcttg tttttagcca aagtttccct tgggaggttt
     1801 ctttgtctgc tctgagactc agctctagaa atataatctg caactgtgac tagcccattt
     1861 tcattaactg aagctgtatc agattcagtc attccatcca tcagaacagc atcttcatca
     1921 gtccttcgtc tacgagaccg cctacgacgg ttagtggaat ggtggctttc ggatgcatca
     1981 gtgtctgcag tctgatctga ttctgtatta tcaagtaagc tgtatggatt gctgtctgga
     2041 tctttgagca cagaactgat ggaggatttg ccaccacgtg gtccaccacg acctcgaccc
     2101 cctgaaacac ttctgcctct tcctcctggg cgtctcctgc tgtcacgctg atgtcggctg
     2161 tctcgatcat cttctcctgc caatgaccaa tcactcagct cgtctttacg ctcagattcc
     2221 gtttcagagg ggttagacag ctcagaattt gtaccataac cggaggtgta attaggtccc
     2281 cgacgacctc tgcctcttcc gctataagac ctagaaccaa tctgtcgcag ctgttcatca
     2341 atctgtaggc gttccattct tagctgttct ggttccttta gataggcaat atgatactct
     2401 agaagaacct gcacatttcc aatgctttct ttagtgccaa caaatacaaa tggaaccata
     2461 ccgtcttctc tgggtaattt attttcattg tccccttcaa ttctcactcg aaccacacca
     2521 gatttgtcca ctatttcttg aataactttg ccattttttc caattacttt tccaacgaga
     2581 ttcctaggaa cctgaataaa atcctccaca aattccaaga aacctctagc cttttttaca
     2641 gcatcagcac tctctccgta gattctgaat gttccagtat cttcatctag ctcaatggcg
     2701 gtaactccag gaaccttcct agcttgctgg atgttactac catgtgttcc tattgccagg
     2761 cccattaaat cttctctcac aacaaattcc tcatgaaaag ctgctgcaag ttgttttgtg
     2821 cattctaaat gcttagtggc ctcttcattt ctggacataa gcatcaactt cgtacgaata
     2881 cttcgcaaat gcatgtcact taagatgttt actctcttca cagttgcttc actggcagac
     2941 agtatcatta gctgtgttgt ttctggatgg taaaaaattc tgcatgctcc tactgctttc
     3001 ttaaaatctt tatgtgcatt ttcattagca cacgcctctc tcaaatcctc aggaacatcc
     3061 actgtgcatt taaagaaggt attttttttg acagttttat tttgattgac aggccgaagt
     3121 cgttcaaatg tgactatttc attgtaagta gcgtcacaag cagcatattc aatgacataa
     3181 aattctcctt tcatcatccg aactttagcc aaccaccacc cacatggctc ttggtcattt
     3241 gctcttgaat atacctctac ttcatctcct tcactaattt ctttttttat atcaggtggt
     3301 ggtggtaatc taacttcatt aaatggaacc tggcgttctg gttgccaatt attttcaaaa
     3361 acaactgtaa gggagtcttc atgaacatct ttgataaatc ccttgtagaa agccccgtta
     3421 gagccgcgaa cctccaccgt cagctccgcc ttgtacagct cgtccatgcc gagagtgatc
     3481 ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg
     3541 ctcagcttgg actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
     3601 atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg
     3661 cggatcttga agttcacctt gatgccgttc ttctgcttgt cggccatgat atagacgttg
     3721 tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg
     3781 atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg
     3841 gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc
     3901 atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc
     3961 acgccgtagg tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg
     4021 cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg
     4081 ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg
     4141 aacagctcct cgcccttgct caccatggtt gctagccagc ttgggtctcc ctatagtgag
     4201 tcgtattaat ttcgataagc cagtaagcag tgggttctct agttagccag agagctctgc
     4261 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
     4321 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
     4381 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
     4441 gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     4501 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     4561 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     4621 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     4681 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     4741 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     4801 ttgattacta ttaataacta gtcaataatc aatgtcaacg cgtatatctg gcccgtacat
     4861 cgcgaagcag cgcaaaacgc ctaaccctaa gcagattctt catgcaattg tcggtcaagc
     4921 cttgccttgt tgtagcttaa attttgctcg cgcactactc agcgacctcc aacacacaag
     4981 cagggagcag atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     5041 ataggggatc gggagatctc ccgatccgtc gacgtcaggt ggcacttttc ggggaaatgt
     5101 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5161 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5221 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5281 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5341 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5401 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5461 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5521 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5581 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5641 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5701 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5761 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5821 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5881 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5941 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6001 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6061 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6121 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6181 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6241 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6301 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6361 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6421 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6481 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6541 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6601 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6661 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6721 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6781 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6841 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6901 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6961 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     7021 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     7081 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     7141 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     7201 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga
     7261 cggtatacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     7321 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     7381 agctgcaata aacaagttgg ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     7441 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     7501 ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc
     7561 cagagtcccg ctcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc
     7621 acctcgacgt cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc
     7681 gc
//
