LOCUS       sequence-458076-        7682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-fusion protein of FXR1 isoform a (N202P).
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-CC1mut (N202P)
FEATURES             Location/Qualifiers
     source          1..7682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1040..1161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1051..1070)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1105..1124
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1335..1934)
                     /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(1418..1438)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1915..1934
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(1947..2276)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1961..2096
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2015..2034)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2261..2281
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2290..2718)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2400..2421)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2612..2631
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2764..2988
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2977..2994
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(4895..5605,5606..5608,5609..5611))
                     /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(4937..4958)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5286..5305
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5545..5566
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5634..5653)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5635..5653)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5698..5901)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5727..5747)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5902..6281
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6453..6472
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6547..6651
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6652..6720,6721..7512)
                     /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(6870..6889)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     1021 cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1081 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1141 ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg
     1201 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     1261 ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa
     1321 ccccagagtc ccgctcaggc accgggcttg cgggtcatgc accaggtgcg cggtccttcg
     1381 ggcacctcga cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg
     1441 ggcgcggagg tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc cactccgggg
     1501 agcacgacgg cgctgcccag acccttgccc tggtggtcgg gcgagacgcc gacggtggcc
     1561 aggaaccacg cgggctcctt gggccggtgc ggcgccagga ggccttccat ctgttgctgc
     1621 gcggccagcc gggaaccgct caactcggcc atgcgcgggc cgatctcggc gaacaccgcc
     1681 cccgcttcga cgctctccgg cgtggtccag accgccaccg cggcgccgtc gtccgcgacc
     1741 cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc
     1801 cgctcgatgt ggcggtccgg atcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg
     1861 gcggcgaggg tgcgtacggc cctggggacg tcgtcgcggg tggcgaggcg caccgtgggc
     1921 ttgtactcgg tcatggttgg gagctttttg caaaagccta ggcctccaaa aaagcctcct
     1981 cactacttct ggaatagctc agaggcagag gcggcctcgg cctctgcata aataaaaaaa
     2041 attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc
     2101 ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg
     2161 cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt
     2221 tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc
     2281 cacagaatta attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2341 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2401 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2461 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     2521 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     2581 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2641 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     2701 cgccgctaca gggcgcgtgg ggataccccc tagagcccca gctggttctt tccgcctcag
     2761 aagccataga gcccaccgca tccccagcat gcctgctatt gtcttcccaa tcctccccct
     2821 tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg
     2881 caatttcctc attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag
     2941 gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagtc gaggctgatc
     3001 agcgggttta aacgggccct ctagactcga gttatgaaac accattcagg actgctgctt
     3061 cttgagtgtt ttcttctttt aaggtattaa tcttttcttc tcctggagta cgctgtagct
     3121 tagaaatgtc atcgccagaa gcactagttg ggccgtttat tgccttttct gaaggaccat
     3181 gctcttcaat cacatctttt gccatttctt tcttgttttt agccaaagtt tcccttggga
     3241 ggtttctttg tctgctctga gactcagctc tagaaatata atctgcaact gtgactagcc
     3301 cattttcatt aactgaagct gtatcagatt cagtcattcc atccatcaga acagcatctt
     3361 catcagtcct tcgtctacga gaccgcctac gacggttagt ggaatggtgg ctttcggatg
     3421 catcagtgtc tgcagtctga tctgattctg tattatcaag taagctgtat ggattgctgt
     3481 ctggatcttt gagcacagaa ctgatggagg atttgccacc acgtggtcca ccacgacctc
     3541 gaccccctga aacacttctg cctcttcctc ctgggcgtct cctgctgtca cgctgatgtc
     3601 ggctgtctcg atcatcttct cctgccaatg accaatcact cagctcgtct ttacgctcag
     3661 attccgtttc agaggggtta gacagctcag aatttgtacc ataaccggag gtgtaattag
     3721 gtccccgacg acctctgcct cttccgctat aagacctaga accaatctgt cgcagctgtt
     3781 catcaatctg taggcgttcc attcttagct gttctacttc ctttagatag gcaatatgat
     3841 actctagaag aacctgcaca tttccaatgc tttctttagt gccaacaaat acaaatggaa
     3901 ccataccgtc ttctctgggt aatttatttt cattgtcccc ttcaattctc actcgaacca
     3961 caccagattt gtccactatt tcttgaataa ctttgccatt ttttccaatt acttttccaa
     4021 cgagattcct aggaacctga ataaaatcct ccacaaattc caagaaacct ctagcctttt
     4081 ttacagcatc agcactctct ccgtagattc tgaatgttcc agtatcttca tctagctcaa
     4141 tggcggtaac tccaggaacc ttcctagctt gctggatgtt actaccatgt gttcctattg
     4201 ccaggcccat taaatcttct ctcacaacaa attcctcatg aaaagctgct gcaagttgtt
     4261 ttgtgcattc taaatgctta gtggcctctt caggtctgga cataagcatc aacttcgtac
     4321 gaatacttcg caaatgcatg tcacttaaga tgtttactct cttcacagtt gcttcactgg
     4381 cagacagtat cattagctgt gttgtttctg gatggtaaaa aattctgcat gctcctactg
     4441 ctttcttaaa atctttatgt gcattttcat tagcacacgc ctctctcaaa tcctcaggaa
     4501 catccactgt gcatttaaag aaggtatttt ttttgacagt tttattttga ttgacaggcc
     4561 gaagtcgttc aaatgtgact atttcattgt aagtagcgtc acaagcagca tattcaatga
     4621 cataaaattc tcctttcatc atccgaactt tagccaacca ccacccacat ggctcttggt
     4681 catttgctct tgaatatacc tctacttcat ctccttcact aatttctttt tttatatcag
     4741 gtggtggtgg taatctaact tcattaaatg gaacctggcg ttctggttgc caattatttt
     4801 caaaaacaac tgtaagggag tcttcatgaa catctttgat aaatcccttg tagaaagccc
     4861 cgttagagcc gcgaacctcc accgtcagct ccgccttgta cagctcgtcc atgccgagag
     4921 tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt
     4981 ctttgctcag cttggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt
     5041 cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt
     5101 tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga
     5161 cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga
     5221 agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg
     5281 cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt
     5341 cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc
     5401 actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg
     5461 tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg
     5521 acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc
     5581 cggtgaacag ctcctcgccc ttgctcacca tggttgctag ccagcttggg tctccctata
     5641 gtgagtcgta ttaatttcga taagccagta agcagtgggt tctctagtta gccagagagc
     5701 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
     5761 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     5821 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     5881 gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca
     5941 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     6001 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     6061 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     6121 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     6181 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     6241 cgtaattgat tactattaat aactagtcaa taatcaatgt caacgcgtat atctggcccg
     6301 tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc aattgtcggt
     6361 caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga cctccaacac
     6421 acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     6481 gcaccatagg ggatcgggag atctcccgat ccgtcgacgt caggtggcac ttttcgggga
     6541 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6601 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6661 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6721 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6781 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6841 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6901 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6961 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     7021 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     7081 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     7141 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     7201 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     7261 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     7321 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     7381 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     7441 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     7501 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     7561 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     7621 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7681 tc
//
