LOCUS       sequence-448477-        7682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-fusion protein of FXR1 isoform a (T236D H237D K299D 
            N300D).
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-KH12mut (T236D H237D K...
FEATURES             Location/Qualifiers
     source          1..7682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(12..215)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(41..61)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        216..595
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     767..786
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        861..965
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(966..1034,1035..1826)
                     /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(1184..1203)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1997..2585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2486..2505
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2739..2756
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2873..2894
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2909..2914,2915..2932,2933..2939)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2947..2963
                     /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     2952..2974
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2971..2987
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2971..2987
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3036..3157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3047..3066)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3101..3120
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3331..3930)
                     /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(3414..3434)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3911..3930
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(3943..4272)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3957..4092
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4011..4030)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     4257..4277
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4286..4714)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4396..4417)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4608..4627
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    4760..4984
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4973..4990
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(6891..7601,7602..7604,7605..7607))
                     /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(6933..6954)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     7282..7301
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7541..7562
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7630..7649)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7631..7649)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gttagccaga gagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc
       61 gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac
      121 aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat
      181 ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg
      241 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
      301 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
      361 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
      421 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
      481 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
      541 tgaactaatg accccgtaat tgattactat taataactag tcaataatca atgtcaacgc
      601 gtatatctgg cccgtacatc gcgaagcagc gcaaaacgcc taaccctaag cagattcttc
      661 atgcaattgt cggtcaagcc ttgccttgtt gtagcttaaa ttttgctcgc gcactactca
      721 gcgacctcca acacacaagc agggagcaga tactggctta actatgcggc atcagagcag
      781 attgtactga gagtgcacca taggggatcg ggagatctcc cgatccgtcg acgtcaggtg
      841 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      901 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      961 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     1021 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     1081 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     1141 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1201 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1261 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1321 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1381 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1441 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1501 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     1561 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     1621 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     1681 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1741 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1801 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1861 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1921 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1981 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     2041 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2101 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     2161 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2221 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2281 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2341 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2401 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2461 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2521 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2581 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2641 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     2701 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     2761 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     2821 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2881 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2941 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     3001 agctctagct agaggtcgac ggtatacaga catgataaga tacattgatg agtttggaca
     3061 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3121 tttatttgta accattataa gctgcaataa acaagttggg gtgggcgaag aactccagca
     3181 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
     3241 acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt
     3301 ggtcggtcat ttcgaacccc agagtcccgc tcaggcaccg ggcttgcggg tcatgcacca
     3361 ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta
     3421 gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc
     3481 cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga
     3541 gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc
     3601 ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat
     3661 ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc
     3721 gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc
     3781 ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg
     3841 gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc
     3901 gaggcgcacc gtgggcttgt actcggtcat ggttgggagc tttttgcaaa agcctaggcc
     3961 tccaaaaaag cctcctcact acttctggaa tagctcagag gcagaggcgg cctcggcctc
     4021 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     4081 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat
     4141 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta
     4201 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc
     4261 taactgacac acattccaca gaattaattc gcgttaaatt tttgttaaat cagctcattt
     4321 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4381 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4441 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     4501 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     4561 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     4621 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     4681 cccgccgcgc ttaatgcgcc gctacagggc gcgtggggat accccctaga gccccagctg
     4741 gttctttccg cctcagaagc catagagccc accgcatccc cagcatgcct gctattgtct
     4801 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
     4861 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
     4921 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
     4981 acagtcgagg ctgatcagcg ggtttaaacg ggccctctag actcgagtta tgaaacacca
     5041 ttcaggactg ctgcttcttg agtgttttct tcttttaagg tattaatctt ttcttctcct
     5101 ggagtacgct gtagcttaga aatgtcatcg ccagaagcac tagttgggcc gtttattgcc
     5161 ttttctgaag gaccatgctc ttcaatcaca tcttttgcca tttctttctt gtttttagcc
     5221 aaagtttccc ttgggaggtt tctttgtctg ctctgagact cagctctaga aatataatct
     5281 gcaactgtga ctagcccatt ttcattaact gaagctgtat cagattcagt cattccatcc
     5341 atcagaacag catcttcatc agtccttcgt ctacgagacc gcctacgacg gttagtggaa
     5401 tggtggcttt cggatgcatc agtgtctgca gtctgatctg attctgtatt atcaagtaag
     5461 ctgtatggat tgctgtctgg atctttgagc acagaactga tggaggattt gccaccacgt
     5521 ggtccaccac gacctcgacc ccctgaaaca cttctgcctc ttcctcctgg gcgtctcctg
     5581 ctgtcacgct gatgtcggct gtctcgatca tcttctcctg ccaatgacca atcactcagc
     5641 tcgtctttac gctcagattc cgtttcagag gggttagaca gctcagaatt tgtaccataa
     5701 ccggaggtgt aattaggtcc ccgacgacct ctgcctcttc cgctataaga cctagaacca
     5761 atctgtcgca gctgttcatc aatctgtagg cgttccattc ttagctgttc tacttccttt
     5821 agataggcaa tatgatactc tagaagaacc tgcacatttc caatgctttc tttagtgcca
     5881 acaaatacaa atggaaccat accgtcttct ctgggtaatt tattttcatt gtccccttca
     5941 attctcactc gaaccacacc agatttgtcc actatttctt gaataacttt gccatcatct
     6001 ccaattactt ttccaacgag attcctagga acctgaataa aatcctccac aaattccaag
     6061 aaacctctag ccttttttac agcatcagca ctctctccgt agattctgaa tgttccagta
     6121 tcttcatcta gctcaatggc ggtaactcca ggaaccttcc tagcttgctg gatgttacta
     6181 ccatcgtctc ctattgccag gcccattaaa tcttctctca caacaaattc ctcatgaaaa
     6241 gctgctgcaa gttgttttgt gcattctaaa tgcttagtgg cctcttcatt tctggacata
     6301 agcatcaact tcgtacgaat acttcgcaaa tgcatgtcac ttaagatgtt tactctcttc
     6361 acagttgctt cactggcaga cagtatcatt agctgtgttg tttctggatg gtaaaaaatt
     6421 ctgcatgctc ctactgcttt cttaaaatct ttatgtgcat tttcattagc acacgcctct
     6481 ctcaaatcct caggaacatc cactgtgcat ttaaagaagg tatttttttt gacagtttta
     6541 ttttgattga caggccgaag tcgttcaaat gtgactattt cattgtaagt agcgtcacaa
     6601 gcagcatatt caatgacata aaattctcct ttcatcatcc gaactttagc caaccaccac
     6661 ccacatggct cttggtcatt tgctcttgaa tatacctcta cttcatctcc ttcactaatt
     6721 tcttttttta tatcaggtgg tggtggtaat ctaacttcat taaatggaac ctggcgttct
     6781 ggttgccaat tattttcaaa aacaactgta agggagtctt catgaacatc tttgataaat
     6841 cccttgtaga aagccccgtt agagccgcga acctccaccg tcagctccgc cttgtacagc
     6901 tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg
     6961 cgcttctcgt tggggtcttt gctcagcttg gactgggtgc tcaggtagtg gttgtcgggc
     7021 agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg
     7081 ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt cttctgcttg
     7141 tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg
     7201 ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc
     7261 tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc
     7321 tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg
     7381 gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc
     7441 acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg
     7501 ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc
     7561 aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggt tgctagccag
     7621 cttgggtctc cctatagtga gtcgtattaa tttcgataag ccagtaagca gtgggttctc
     7681 ta
//
