LOCUS       sequence-448481-        7682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-fusion protein of FXR1 isoform a (T236D H237D).
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-FXR1-N1-KH1mut (T236D H237D)
FEATURES             Location/Qualifiers
     source          1..7682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(25..44)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      838..1426
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1327..1346
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1580..1597
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1714..1735
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1750..1755,1756..1773,1774..1780)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1788..1804
                     /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     1793..1815
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1812..1828
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1812..1828
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1877..1998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1888..1907)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1942..1961
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2172..2771)
                     /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(2255..2275)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2752..2771
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2784..3113)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2798..2933
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2852..2871)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3098..3118
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3127..3555)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3237..3258)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3449..3468
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3601..3825
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3814..3831
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(5732..6442,6443..6445,6446..6448))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5774..5795)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     6123..6142
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6382..6403
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6471..6490)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6472..6490)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6535..6738)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6564..6584)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6739..7118
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7290..7309
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7384..7488
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7489..7557,7558..7682,1..667)
                     /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"
ORIGIN
        1 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
       61 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      121 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      181 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
      241 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
      301 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
      361 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
      421 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
      481 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
      541 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
      601 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
      661 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
      721 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
      781 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
      841 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
      901 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
      961 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1021 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1081 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1141 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1201 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     1261 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     1321 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     1381 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     1441 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     1501 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     1561 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     1621 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     1681 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     1741 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     1801 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga
     1861 cggtatacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     1921 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     1981 agctgcaata aacaagttgg ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     2041 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     2101 ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc
     2161 cagagtcccg ctcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc
     2221 acctcgacgt cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc
     2281 gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc
     2341 acgacggcgc tgcccagacc cttgccctgg tggtcgggcg agacgccgac ggtggccagg
     2401 aaccacgcgg gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg
     2461 gccagccggg aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc
     2521 gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac
     2581 accttgccga tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc
     2641 tcgatgtggc ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg
     2701 gcgagggtgc gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg
     2761 tactcggtca tggttgggag ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac
     2821 tacttctgga atagctcaga ggcagaggcg gcctcggcct ctgcataaat aaaaaaaatt
     2881 agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga
     2941 gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     3001 gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc
     3061 atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca cacattccac
     3121 agaattaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     3181 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     3241 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     3301 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     3361 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     3421 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3481 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3541 cgctacaggg cgcgtgggga taccccctag agccccagct ggttctttcc gcctcagaag
     3601 ccatagagcc caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc
     3661 tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa
     3721 tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca
     3781 cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgag gctgatcagc
     3841 gggtttaaac gggccctcta gactcgagtt atgaaacacc attcaggact gctgcttctt
     3901 gagtgttttc ttcttttaag gtattaatct tttcttctcc tggagtacgc tgtagcttag
     3961 aaatgtcatc gccagaagca ctagttgggc cgtttattgc cttttctgaa ggaccatgct
     4021 cttcaatcac atcttttgcc atttctttct tgtttttagc caaagtttcc cttgggaggt
     4081 ttctttgtct gctctgagac tcagctctag aaatataatc tgcaactgtg actagcccat
     4141 tttcattaac tgaagctgta tcagattcag tcattccatc catcagaaca gcatcttcat
     4201 cagtccttcg tctacgagac cgcctacgac ggttagtact gtgatgagat tcgctggcat
     4261 cagtgtctgc agtctgatct gattctgtat tatcaagtaa gctgtatgga ttgctgtctg
     4321 gatctttgag cacagaactg atggaggatt tgccaccacg tggtccacca cgacctcgac
     4381 cccctgaaac acttctgcct cttcctcctg ggcgtctcct gctgtcacgc tgatgtcggc
     4441 tgtctcgatc atcttctcct gccaatgacc aatcactcag ctcgtcttta cgctcagatt
     4501 ccgtttcaga ggggttagac agctcagaat ttgtaccata accggaggtg taattaggtc
     4561 cccgacgacc tctgcctctt ccgctataag acctagaacc aatctgtcgc agctgttcat
     4621 caatctgtag gcgttccatt cttagctgtt ctacttcctt tagataggca atatgatact
     4681 ctagaagaac ctgcacattt ccaatgcttt ctttagtgcc aacaaataca aatggaacca
     4741 taccgtcttc tctgggtaat ttattttcat tgtccccttc aattctcact cgaaccacac
     4801 cagatttgtc cactatttct tgaataactt tgccattttt tccaattact tttccaacga
     4861 gattcctagg aacctgaata aaatcctcca caaattccaa gaaacctcta gcctttttta
     4921 cagcatcagc actctctccg tagattctga atgttccagt atcttcatct agctcaatgg
     4981 cggtaactcc aggaaccttc ctagcttgct ggatgttact accatcgtct cctattgcca
     5041 ggcccattaa atcttctctc acaacaaatt cctcatgaaa agctgctgca agttgttttg
     5101 tgcattctaa atgcttagtg gcctcttcat ttctggacat aagcatcaac ttcgtacgaa
     5161 tacttcgcaa atgcatgtca cttaagatgt ttactctctt cacagttgct tcactggcag
     5221 acagtatcat tagctgtgtt gtttctggat ggtaaaaaat tctgcatgct cctactgctt
     5281 tcttaaaatc tttatgtgca ttttcattag cacacgcctc tctcaaatcc tcaggaacat
     5341 ccactgtgca tttaaagaag gtattttttt tgacagtttt attttgattg acaggccgaa
     5401 gtcgttcaaa tgtgactatt tcattgtaag tagcgtcaca agcagcatat tcaatgacat
     5461 aaaattctcc tttcatcatc cgaactttag ccaaccacca cccacatggc tcttggtcat
     5521 ttgctcttga atatacctct acttcatctc cttcactaat ttcttttttt atatcaggtg
     5581 gtggtggtaa tctaacttca ttaaatggaa cctggcgttc tggttgccaa ttattttcaa
     5641 aaacaactgt aagggagtct tcatgaacat ctttgataaa tcccttgtag aaagccccgt
     5701 tagagccgcg aacctccacc gtcagctccg ccttgtacag ctcgtccatg ccgagagtga
     5761 tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt
     5821 tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc
     5881 cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt
     5941 ggcggatctt gaagttcgcc ttgatgccgt tcttctgctt gtcggccatg atatagacgt
     6001 tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt
     6061 cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc
     6121 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg
     6181 gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact
     6241 gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg
     6301 tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccagaca
     6361 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg
     6421 tgaacagctc ctcgcccttg ctcaccatgg ttgctagcca gcttgggtct ccctatagtg
     6481 agtcgtatta atttcgataa gccagtaagc agtgggttct ctagttagcc agagagctct
     6541 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
     6601 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
     6661 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     6721 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     6781 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     6841 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     6901 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     6961 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     7021 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     7081 aattgattac tattaataac tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac
     7141 atcgcgaagc agcgcaaaac gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa
     7201 gccttgcctt gttgtagctt aaattttgct cgcgcactac tcagcgacct ccaacacaca
     7261 agcagggagc agatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     7321 ccatagggga tcgggagatc tcccgatccg tcgacgtcag gtggcacttt tcggggaaat
     7381 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     7441 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     7501 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     7561 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     7621 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     7681 cc
//
