LOCUS       sequence-448465-        7682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-fusion protein of FXR1 isoform a (N202P 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-Ccmut (N202P-V361P)
FEATURES             Location/Qualifiers
     source          1..7682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..127
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(128..196,197..988)
                     /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(346..365)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1159..1747
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1648..1667
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1901..1918
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2035..2056
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2071..2076,2077..2094,2095..2101)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2109..2125
                     /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     2114..2136
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2133..2149
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2133..2149
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2198..2319
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2209..2228)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2263..2282
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2493..3092)
                     /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(2576..2596)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3073..3092
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(3105..3434)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3119..3254
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3173..3192)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3419..3439
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3448..3876)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3558..3579)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3770..3789
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3922..4146
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4135..4152
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(6053..6763,6764..6766,6767..6769))
                     /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(6095..6116)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     6444..6463
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6703..6724
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6792..6811)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6793..6811)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6856..7059)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6885..6905)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7060..7439
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7611..7630
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
       61 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      121 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
      181 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
      241 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
      301 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
      361 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
      421 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
      481 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
      541 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
      601 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
      661 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
      721 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
      781 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
      841 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
      901 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
      961 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     1021 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1081 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1141 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1201 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1261 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     1321 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1381 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1441 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1501 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     1561 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     1621 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     1681 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     1741 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     1801 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     1861 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     1921 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     1981 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     2041 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     2101 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     2161 caagctctag ctagaggtcg acggtataca gacatgataa gatacattga tgagtttgga
     2221 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2281 gctttatttg taaccattat aagctgcaat aaacaagttg gggtgggcga agaactccag
     2341 catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc
     2401 caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc
     2461 ttggtcggtc atttcgaacc ccagagtccc gctcaggcac cgggcttgcg ggtcatgcac
     2521 caggtgcgcg gtccttcggg cacctcgacg tcggcggtga cggtgaagcc gagccgctcg
     2581 tagaagggga ggttgcgggg cgcggaggtc tccaggaagg cgggcacccc ggcgcgctcg
     2641 gccgcctcca ctccggggag cacgacggcg ctgcccagac ccttgccctg gtggtcgggc
     2701 gagacgccga cggtggccag gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg
     2761 ccttccatct gttgctgcgc ggccagccgg gaaccgctca actcggccat gcgcgggccg
     2821 atctcggcga acaccgcccc cgcttcgacg ctctccggcg tggtccagac cgccaccgcg
     2881 gcgccgtcgt ccgcgaccca caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt
     2941 tcttgcagct cggtgacccg ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg
     3001 gggtagtcgg cgaacgcggc ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg
     3061 gcgaggcgca ccgtgggctt gtactcggtc atggttggga gctttttgca aaagcctagg
     3121 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggcagaggc ggcctcggcc
     3181 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     3241 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
     3301 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
     3361 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     3421 cctaactgac acacattcca cagaattaat tcgcgttaaa tttttgttaa atcagctcat
     3481 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3541 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3601 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3661 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3721 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3781 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3841 cacccgccgc gcttaatgcg ccgctacagg gcgcgtgggg atacccccta gagccccagc
     3901 tggttctttc cgcctcagaa gccatagagc ccaccgcatc cccagcatgc ctgctattgt
     3961 cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc
     4021 tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac
     4081 cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag
     4141 gcacagtcga ggctgatcag cgggtttaaa cgggccctct agactcgagt tatgaaacac
     4201 cattcaggac tgctgcttct tgagtgtttt cttcttttaa ggtattaatc ttttcttctc
     4261 ctggagtacg ctgtagctta gaaatgtcat cgccagaagc actagttggg ccgtttattg
     4321 ccttttctga aggaccatgc tcttcaatca catcttttgc catttctttc ttgtttttag
     4381 ccaaagtttc ccttgggagg tttctttgtc tgctctgaga ctcagctcta gaaatataat
     4441 ctgcaactgt gactagccca ttttcattaa ctgaagctgt atcagattca gtcattccat
     4501 ccatcagaac agcatcttca tcagtccttc gtctacgaga ccgcctacga cggttagtgg
     4561 aatggtggct ttcggatgca tcagtgtctg cagtctgatc tgattctgta ttatcaagta
     4621 agctgtatgg attgctgtct ggatctttga gcacagaact gatggaggat ttgccaccac
     4681 gtggtccacc acgacctcga ccccctgaaa cacttctgcc tcttcctcct gggcgtctcc
     4741 tgctgtcacg ctgatgtcgg ctgtctcgat catcttctcc tgccaatgac caatcactca
     4801 gctcgtcttt acgctcagat tccgtttcag aggggttaga cagctcagaa tttgtaccat
     4861 aaccggaggt gtaattaggt ccccgacgac ctctgcctct tccgctataa gacctagaac
     4921 caatctgtcg cagctgttca tcaatctgta ggcgttccat tcttagctgt tctggttcct
     4981 ttagataggc aatatgatac tctagaagaa cctgcacatt tccaatgctt tctttagtgc
     5041 caacaaatac aaatggaacc ataccgtctt ctctgggtaa tttattttca ttgtcccctt
     5101 caattctcac tcgaaccaca ccagatttgt ccactatttc ttgaataact ttgccatttt
     5161 ttccaattac ttttccaacg agattcctag gaacctgaat aaaatcctcc acaaattcca
     5221 agaaacctct agcctttttt acagcatcag cactctctcc gtagattctg aatgttccag
     5281 tatcttcatc tagctcaatg gcggtaactc caggaacctt cctagcttgc tggatgttac
     5341 taccatgtgt tcctattgcc aggcccatta aatcttctct cacaacaaat tcctcatgaa
     5401 aagctgctgc aagttgtttt gtgcattcta aatgcttagt ggcctcttca ggtctggaca
     5461 taagcatcaa cttcgtacga atacttcgca aatgcatgtc acttaagatg tttactctct
     5521 tcacagttgc ttcactggca gacagtatca ttagctgtgt tgtttctgga tggtaaaaaa
     5581 ttctgcatgc tcctactgct ttcttaaaat ctttatgtgc attttcatta gcacacgcct
     5641 ctctcaaatc ctcaggaaca tccactgtgc atttaaagaa ggtatttttt ttgacagttt
     5701 tattttgatt gacaggccga agtcgttcaa atgtgactat ttcattgtaa gtagcgtcac
     5761 aagcagcata ttcaatgaca taaaattctc ctttcatcat ccgaacttta gccaaccacc
     5821 acccacatgg ctcttggtca tttgctcttg aatatacctc tacttcatct ccttcactaa
     5881 tttctttttt tatatcaggt ggtggtggta atctaacttc attaaatgga acctggcgtt
     5941 ctggttgcca attattttca aaaacaactg taagggagtc ttcatgaaca tctttgataa
     6001 atcccttgta gaaagccccg ttagagccgc gaacctccac cgtcagctcc gccttgtaca
     6061 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
     6121 cgcgcttctc gttggggtct ttgctcagct tggactgggt gctcaggtag tggttgtcgg
     6181 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
     6241 cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct
     6301 tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga
     6361 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
     6421 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc
     6481 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
     6541 cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg
     6601 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat
     6661 cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     6721 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg gttgctagcc
     6781 agcttgggtc tccctatagt gagtcgtatt aatttcgata agccagtaag cagtgggttc
     6841 tctagttagc cagagagctc tgcttatata gacctcccac cgtacacgcc taccgcccat
     6901 ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca
     6961 aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg
     7021 ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa
     7081 tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca
     7141 ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg
     7201 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
     7261 ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt
     7321 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg
     7381 gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtca
     7441 acgcgtatat ctggcccgta catcgcgaag cagcgcaaaa cgcctaaccc taagcagatt
     7501 cttcatgcaa ttgtcggtca agccttgcct tgttgtagct taaattttgc tcgcgcacta
     7561 ctcagcgacc tccaacacac aagcagggag cagatactgg cttaactatg cggcatcaga
     7621 gcagattgta ctgagagtgc accatagggg atcgggagat ctcccgatcc gtcgacgtca
     7681 gg
//
