LOCUS       sequence-448631-        7682 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express EGFP-fusion protein of FXR1 isoform a (CCswap).
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 Apr 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-Ccswap
FEATURES             Location/Qualifiers
     source          1..7682
                     /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(905..907,908..910,911..1621)
                     /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(950..971)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1211..1230)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1558..1579
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3522..3539)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3528..3752
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3798..4226
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3885..3904)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4095..4116
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4235..4255)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4240..4569
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4420..4555
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4482..4501
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4582..5181
                     /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(4582..4601)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5078..5098
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5355..5476
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5392..5411)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5446..5465
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5525..5541)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5525..5541)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5538..5560)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5549..5565
                     /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(5573..5579,5580..5597,5598..5603))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5618..5639
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5756..5773)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5927..6515)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6007..6026)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6686..7477,7478..7546))
                     /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     7309..7328
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7547..7651)
                     /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 cgtgtccggc gagggcgagg gcgatgccac
     1021 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1081 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     1141 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1201 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1261 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1321 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1381 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1441 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1501 ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     1561 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1621 ggcggagctg acggtggagg ttcgcggctc taacggggct ttctacaagg gatttatcaa
     1681 agatgttcat gaagactccc ttacagttgt ttttgaaaat aattggcaac cagaacgcca
     1741 ggttccattt aatgaagtta gattaccacc accacctgat ataaaaaaag aaattagtga
     1801 aggagatgaa gtagaggtat attcaagagc aaatgaccaa gagccatgtg ggtggtggtt
     1861 ggctaaagtt cggatgatga aaggagaatt ttatgtcatt gaatatgctg cttgtgacgc
     1921 tacttacaat gaaatagtca catttgaacg acttcggcct gtcaatcaaa ataaaactgt
     1981 caaaaaaaat accttcttta aatgcacagt ggatgttcct gaggatttga gagaggcgtg
     2041 tgctaatgaa aatgcacata aagattttaa gaaagcagta ggagcatgca gaatttttta
     2101 ccatccagaa acaacacagc taatgatact gtctgccagt gaagcaactg tgaagagagt
     2161 aaacatctta agtgacatgc atttgcgaag tattcgtacg aagttgatgc ttatgtccag
     2221 acatattgcc tatctaaagg aagtagaaca gctaagaatg gaacgcctac agattgatga
     2281 acagtttgtt gtgagagaag atttaatggg cctggcaata ggaacacatg gtagtaacat
     2341 ccagcaagct aggaaggttc ctggagttac cgccattgag ctagatgaag atactggaac
     2401 attcagaatc tacggagaga gtgctgatgc tgtaaaaaag gctagaggtt tcttggaatt
     2461 tgtggaggat tttattcagg ttcctaggaa tctcgttgga aaagtaattg gaaaaaatgg
     2521 caaagttatt caagaaatag tggacaaatc tggtgtggtt cgagtgagaa ttgaagggga
     2581 caatgaaaat aaattaccca gagaagacgg tatggttcca tttgtatttg ttggcactaa
     2641 agaaagcatt ggaaatgtgc aggttcttct agagtatcat attgcctatc taaaggaaaa
     2701 tgaagaggcc actaagcatt tagaatgcac aaaacaactt gcagcagctt ttggttctag
     2761 gtcttatagc ggaagaggca gaggtcgtcg gggacctaat tacacctccg gttatggtac
     2821 aaattctgag ctgtctaacc cctctgaaac ggaatctgag cgtaaagacg agctgagtga
     2881 ttggtcattg gcaggagaag atgatcgaga cagccgacat cagcgtgaca gcaggagacg
     2941 cccaggagga agaggcagaa gtgtttcagg gggtcgaggt cgtggtggac cacgtggtgg
     3001 caaatcctcc atcagttctg tgctcaaaga tccagacagc aatccataca gcttacttga
     3061 taatacagaa tcagatcaga ctgcagacac tgatgcatcc gaaagccacc attccactaa
     3121 ccgtcgtagg cggtctcgta gacgaaggac tgatgaagat gctgttctga tggatggaat
     3181 gactgaatct gatacagctt cagttaatga aaatgggcta gtcacagttg cagattatat
     3241 ttctagagct gagtctcaga gcagacaaag aaacctccca agggaaactt tggctaaaaa
     3301 caagaaagaa atggcaaaag atgtgattga agagcatggt ccttcagaaa aggcaataaa
     3361 cggcccaact agtgcttctg gcgatgacat ttctaagcta cagcgtactc caggagaaga
     3421 aaagattaat accttaaaag aagaaaacac tcaagaagca gcagtcctga atggtgtttc
     3481 ataactcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     3541 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3601 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3661 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3721 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     3781 ctctatgggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3841 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3901 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3961 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4021 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4081 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4141 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4201 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4261 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4321 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4381 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4441 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4501 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4561 ttttgcaaaa agctcccaac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     4621 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     4681 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     4741 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     4801 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     4861 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     4921 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     4981 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     5041 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     5101 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     5161 acccgcaagc ccggtgcctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5221 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5281 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5341 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5401 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5461 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5521 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5581 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5641 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5701 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5761 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5821 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5881 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5941 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6001 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6061 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6121 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6181 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6241 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6301 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6361 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6421 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6481 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6541 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6601 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6661 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6721 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6781 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6841 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6901 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6961 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7021 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7081 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7141 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7201 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7261 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7321 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7381 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7441 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7501 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7561 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7621 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7681 tc
//
