LOCUS       sequence-453904-        7703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express EGFP-fusion protein of FXR1 isoform b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7703)
  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 7703)
  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-EGFP-FXR1-isoform b
FEATURES             Location/Qualifiers
     source          1..7703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1139..1141,1142..1144,1145..1855)
                     /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(1445..1464)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1792..1813
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3543..3560)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3549..3773
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3819..4247
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3906..3925)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4116..4137
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4256..4276)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4261..4590
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4441..4576
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4503..4522
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4603..5202
                     /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(4603..4622)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5099..5119
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5376..5497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5413..5432)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5467..5486
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5546..5562)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5546..5562)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5559..5581)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5570..5586
                     /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)."
     promoter        complement(join(5594..5600,5601..5618,5619..5624))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5639..5660
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5777..5794)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5948..6536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6028..6047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6707..7498,7499..7567))
                     /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     7330..7349
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7568..7672)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 aaccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      961 ggacggcgac gtaaacggcc acaagttcag cgtgtctggc gagggcgagg gcgatgccac
     1021 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1081 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgacmmcat
     1141 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1201 cgacgtaaac ggccacaagt tcagcgtgtc tggcgagggc gagggcgatg ccacctacgg
     1261 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1321 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     1381 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1441 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1501 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1561 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1621 catcaaggcg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1681 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1741 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1801 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaaggcgga
     1861 gctgacggtg gaggttcgcg gctctaacgg ggctttctac aagggattta tcaaagatgt
     1921 tcatgaagac tcccttacag ttgtttttga aaataattgg caaccagaac gccaggttcc
     1981 atttaatgaa gttagattac caccaccacc tgatataaaa aaagaaatta gtgaaggaga
     2041 tgaagtagag gtatattcaa gagcaaatga ccaagagcca tgtgggtggt ggttggctaa
     2101 agttcggatg atgaaaggag aattttatgt cattgaatat gctgcttgtg acgctactta
     2161 caatgaaata gtcacatttg aacgacttcg gcctgtcaat caaaataaaa ctgtcaaaaa
     2221 aaataccttc tttaaatgca cagtggatgt tcctgaggat ttgagagagg cgtgtgctaa
     2281 tgaaaatgca cataaagatt ttaagaaagc agtaggagca tgcagaattt tttaccatcc
     2341 agaaacaaca cagctaatga tactgtctgc cagtgaagca actgtgaaga gagtaaacat
     2401 cttaagtgac atgcatttgc gaagtattcg tacgaagttg atgcttatgt ccagaaatga
     2461 agaggccact aagcatttag aatgcacaaa acaacttgca gcagcttttc atgaggaatt
     2521 tgttgtgaga gaagatttaa tgggcctggc aataggaaca catggtagta acatccagca
     2581 agctaggaag gttcctggag ttaccgccat tgagctagat gaagatactg gaacattcag
     2641 aatctacgga gagagtgctg atgctgtaaa aaaggctaga ggtttcttgg aatttgtgga
     2701 ggattttatt caggttccta ggaatctcgt tggaaaagta attggaaaaa atggcaaagt
     2761 tattcaagaa atagtggaca aatctggtgt ggttcgagtg agaattgaag gggacaatga
     2821 aaataaatta cccagagaag acggtatggt tccatttgta tttgttggca ctaaagaaag
     2881 cattggaaat gtgcaggttc ttctagagta tcatattgcc tatctaaagg aagtagaaca
     2941 gctaagaatg gaacgcctac agattgatga acagctgcga cagattggtt ctaggtctta
     3001 tagcggaaga ggcagaggtc gtcggggacc taattacacc tccggttatg gtacaaattc
     3061 tgagctgtct aacccctctg aaacggaatc tgagcgtaaa gacgagctga gtgattggtc
     3121 attggcagga gaagatgatc gagacagccg acatcagcgt gacagcagga gacgcccagg
     3181 aggaagaggc agaagtgttt cagggggtcg aggtcgtggt ggaccacgtg gtggcaaatc
     3241 ctccatcagt tctgtgctca aagatccaga cagcaatcca tacagcttac ttgataatac
     3301 agaatcagat cagactgcag acactgatgc cagcgaatct catcacagta ctaaccgtcg
     3361 taggcggtct cgtagacgaa ggactgatga agatgctgtt ctgatggatg gaatgactga
     3421 atctgataca gcttcagtta atgaaaatgg gctaggcaaa agatgtgatt gagaattctg
     3481 cagatatcca gcacagtggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc
     3541 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3601 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3661 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3721 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3781 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3841 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3901 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3961 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4021 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4081 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4141 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     4201 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4261 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4321 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4381 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4441 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4501 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4561 ggcttttttg gaggcctagg cttttgcaaa aagctcccaa ccatgaccga gtacaagccc
     4621 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     4681 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     4741 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     4801 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     4861 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     4921 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     4981 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     5041 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     5101 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     5161 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagcgggact ctggggttcg
     5221 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5281 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5341 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5401 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5461 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5521 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5581 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5641 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5701 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5761 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5821 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5881 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5941 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6001 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6061 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6121 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6181 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6241 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6301 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6361 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6421 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6481 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6541 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6601 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6661 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6721 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6781 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6841 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6901 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6961 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7021 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7081 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7141 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7201 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7261 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7321 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7381 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7441 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7501 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7561 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7621 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7681 cccgaaaagt gccacctgac gtc
//
