LOCUS       sequence-458067-        7916 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express EGFP-fusion protein of FXR1 isoform a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7916)
  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 7916)
  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-FXR1-isoform a
FEATURES             Location/Qualifiers
     source          1..7916
                     /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="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(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(3756..3773)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3762..3986
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4032..4460
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4119..4138)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4329..4350
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4469..4489)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4474..4803
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4654..4789
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4716..4735
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4816..5415
                     /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(4816..4835)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5312..5332
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5589..5710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5626..5645)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5680..5699
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5759..5775)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5759..5775)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5772..5794)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5783..5799
                     /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(5807..5813,5814..5831,5832..5837))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5852..5873
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5990..6007)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6161..6749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6241..6260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6920..7711,7712..7780))
                     /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     7543..7562
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7781..7885)
                     /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 tagcaaccat
     1141 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1201 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc 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 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1681 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1741 cctgagcacc cagtccaagc 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 gctagtcaca gttgcagatt atatttctag
     3481 agctgagtct cagagcagac aaagaaacct cccaagggaa actttggcta aaaacaagaa
     3541 agaaatggca aaagatgtga ttgaagagca tggtccttca gaaaaggcaa taaacggccc
     3601 aactagtgct tctggcgatg acatttctaa gctacagcgt actccaggag aagaaaagat
     3661 taatacctta aaagaagaaa acactcaaga agcagcagtc ctgaatggtg tttcataact
     3721 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3781 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3841 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3901 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3961 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     4021 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4081 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4141 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4201 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4261 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4321 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4381 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4441 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4501 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4561 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4621 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4681 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4741 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4801 aaaaagctcc caaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     4861 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     4921 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     4981 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     5041 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     5101 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     5161 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     5221 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     5281 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     5341 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     5401 aagcccggtg cctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5461 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5521 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5581 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5641 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5701 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5761 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5821 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5881 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5941 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6001 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6061 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6121 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6181 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6241 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6301 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6361 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6421 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6481 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6541 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6601 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6661 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6721 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6781 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6841 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6901 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6961 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7021 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7081 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7141 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7201 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7261 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7321 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7381 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7441 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7501 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7561 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7621 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7681 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7741 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7801 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7861 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
