LOCUS       sequence-453527-        6550 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express mEGFP-fusion protein of FXR1 isoform a (FXR1 fragment 
            (379-621)).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6550)
  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 6550)
  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-EGFP-FXR1-C (379-621)
FEATURES             Location/Qualifiers
     source          1..6550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      593..1181
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1082..1101
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1335..1352
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1469..1490
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1505..1510,1511..1528,1529..1535)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1543..1559
                     /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     1548..1570
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1567..1583
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1567..1583
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1632..1753
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1643..1662)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1697..1716
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1927..2526)
                     /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(2010..2030)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2507..2526
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2539..2868)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2553..2688
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2607..2626)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2853..2873
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2882..3310)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2992..3013)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3204..3223
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3356..3580
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3569..3586
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(4355..5065,5066..5068,5069..5071))
                     /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(4397..4418)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4746..4765
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5005..5026
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5094..5113)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5095..5113)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5158..5361)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5187..5207)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5362..5741
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5913..5932
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6007..6111
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6112..6180,6181..6550,1..422)
                     /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(6330..6349)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
       61 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
      121 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
      181 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
      241 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
      301 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
      361 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
      421 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
      481 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
      541 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
      601 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
      661 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
      721 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
      781 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
      841 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      901 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      961 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     1021 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     1081 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     1141 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     1201 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     1261 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     1321 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     1381 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     1441 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     1501 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     1561 ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag gtcgacggta
     1621 tacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     1681 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     1741 caataaacaa gttggggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca
     1801 tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg
     1861 aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag
     1921 tcccgctcag gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc
     1981 gacgtcggcg gtgacggtga agccgagccg ctcgtagaag gggaggttgc ggggcgcgga
     2041 ggtctccagg aaggcgggca ccccggcgcg ctcggccgcc tccactccgg ggagcacgac
     2101 ggcgctgccc agacccttgc cctggtggtc gggcgagacg ccgacggtgg ccaggaacca
     2161 cgcgggctcc ttgggccggt gcggcgccag gaggccttcc atctgttgct gcgcggccag
     2221 ccgggaaccg ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc
     2281 gacgctctcc ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga cccacacctt
     2341 gccgatgtcg agcccgacgc gcgtgaggaa gagttcttgc agctcggtga cccgctcgat
     2401 gtggcggtcc ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag
     2461 ggtgcgtacg gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc
     2521 ggtcatggtt gggagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     2581 ctggaatagc tcagaggcag aggcggcctc ggcctctgca taaataaaaa aaattagtca
     2641 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     2701 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     2761 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
     2821 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagaat
     2881 taattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2941 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3001 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3061 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3121 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3181 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3241 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3301 cagggcgcgt ggggataccc cctagagccc cagctggttc tttccgcctc agaagccata
     3361 gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc
     3421 tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc
     3481 tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg
     3541 gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcgggtt
     3601 taaacgggcc ctctagactc gagttatgaa acaccatcag gactgctgct tcttgagtgt
     3661 tttcttcttt taaggtatta atcttttctt ctcctggagt acgctgtagc ttagaaatgt
     3721 catcgccaga agcactagtt gggccgttta ttgccttttc tgaaggacca tgctcttcaa
     3781 tcacatcttt tgccatttct ttcttgtttt tagccaaagt ttcccttggg aggtttcttt
     3841 gtctgctctg agactcagct ctagaaatat aatctgcaac tgtgactagc ccattttcat
     3901 taactgaagc tgtatcagat tcagtcattc catccatcag aacagcatct tcatcagtcc
     3961 ttcgtctacg agaccgccta cgacggttag tactgtgatg agattcgctg gcatcagtgt
     4021 ctgcagtctg atctgattct gtattatcaa gtaagctgta tggattgctg tctggatctt
     4081 tgagcacaga actgatggag gatttgccac cacgtggtcc accacgacct cgaccccctg
     4141 aaacacttct gcctcttcct cctgggcgtc tcctgctgtc acgctgatgt cggctgtctc
     4201 gatcatcttc tcctgccaat gaccaatcac tcagctcgtc tttacgctca gattccgttt
     4261 cagaggggtt agacagctca gaatttgtac cataaccgga ggtgtaatta ggtccccgac
     4321 gacctctgcc tcttccgcta taagacctag aacccttgta cagctcgtcc atgccgagag
     4381 tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt
     4441 ctttgctcag ggcggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt
     4501 cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt
     4561 tgtggcggat cttgaagttc gccttgatgc cgttcttctg cttgtcggcc atgatataga
     4621 cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga
     4681 agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg
     4741 cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt
     4801 cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc
     4861 actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg
     4921 tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccag
     4981 acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc
     5041 cggtgaacag ctcctcgccc ttgctcacca tggttgctag ccagcttggg tctccctata
     5101 gtgagtcgta ttaatttcga taagccagta agcagtgggt tctctagtta gccagagagc
     5161 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
     5221 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     5281 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     5341 gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca
     5401 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     5461 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     5521 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     5581 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     5641 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     5701 cgtaattgat tactattaat aactagtcaa taatcaatgt caacgcgtat atctggcccg
     5761 tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc aattgtcggt
     5821 caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga cctccaacac
     5881 acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     5941 gcaccatagg ggatcgggag atctcccgat ccgtcgacgt caggtggcac ttttcgggga
     6001 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6061 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6121 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6181 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6241 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6301 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6361 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6421 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6481 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6541 aaggagctaa
//
