LOCUS       sequence-392973-        8023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human FXR1 protein in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8023)
  AUTHORS   Edwards M, Xu M, Joseph S
  TITLE     A simple procedure for bacterial expression and purification of the 
            fragile X protein family.
  JOURNAL   Sci Rep. 2020 Sep 28;10(1):15858. doi: 10.1038/s41598-020-72984-7.
  PUBMED    32985615
REFERENCE   2  (bases 1 to 8023)
  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: "Sci Rep."; 
            date: "2020-09-28"; volume: "10(1):15858. doi"; pages: " 
            10.1038/s41598-020-72984-7"
COMMENT     Sequence Label: His6_MBP_FXR1P
FEATURES             Location/Qualifiers
     source          1..8023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      8..596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     497..516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     750..767
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    782..921
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(847..866)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1007..1029
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1023..1214)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2023..3105)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     3067..3086
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3106..3183)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3389..3408
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3496..3515
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3496..3514
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3515..3539
                     /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)."
     RBS             3554..3576
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3587..3604
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3629..4726
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      4583..4592
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4700..4723
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             4748..4768
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             6455..6472
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6525..6543)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6539..6586
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(6710..6729)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6896..6999
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7000..7068,7069..7860)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7218..7237)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
       61 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      121 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
      181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
      541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
      601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
      661 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
      721 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
      781 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac
      841 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
      901 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
      961 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     1021 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     1081 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     1141 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     1201 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     1261 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     1321 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     1381 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     1441 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
     1501 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     1561 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
     1621 aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt
     1681 ggggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca
     1741 gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc
     1801 atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc
     1861 tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc
     1921 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     1981 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     2041 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2101 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     2161 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     2221 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     2281 atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
     2341 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     2401 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     2461 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     2521 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     2581 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     2641 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     2701 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     2761 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     2821 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     2881 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     2941 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     3001 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     3061 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     3121 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     3181 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta
     3241 ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca
     3301 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     3361 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     3421 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatcgatc
     3481 tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
     3541 ctctagaaat aattttgttt aactttaaga aggagatata catatgcacc atcatcatca
     3601 tcattcttct ggtgtagatc tgggtaccaa aatcgaagaa ggtaaactgg taatctggat
     3661 taacggcgat aaaggctata acggtctcgc tgaagtcggt aagaaattcg agaaagatac
     3721 cggaattaaa gtcaccgttg agcatccgga taaactggaa gagaaattcc cacaggttgc
     3781 ggcaactggc gatggccctg acattatctt ctgggcacac gaccgctttg gtggctacgc
     3841 tcaatctggc ctgttggctg aaatcacccc ggacaaagcg ttccaggaca agctgtatcc
     3901 gtttacctgg gatgccgtac gttacaacgg caagctgatt gcttacccga tcgctgttga
     3961 agcgttatcg ctgatttata acaaagatct gctgccgaac ccgccaaaaa cctgggaaga
     4021 gatcccggcg ctggataaag aactgaaagc gaaaggtaag agcgcgctga tgttcaacct
     4081 gcaagaaccg tacttcacct ggccgctgat tgctgctgac gggggttatg cgttcaagta
     4141 tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat aacgctggcg cgaaagcggg
     4201 tctgaccttc ctggttgacc tgattaaaaa caaacacatg aatgcagaca ccgattactc
     4261 catcgcagaa gctgccttta ataaaggcga aacagcgatg accatcaacg gcccgtgggc
     4321 atggtccaac atcgacacca gcaaagtgaa ttatggtgta acggtactgc cgaccttcaa
     4381 gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca ggtattaacg ccgccagtcc
     4441 gaacaaagag ctggcaaaag agttcctcga aaactatctg ctgactgatg aaggtctgga
     4501 agcggttaat aaagacaaac cgctgggtgc cgtagcgctg aagtcttacg aggaagagtt
     4561 ggcgaaagat ccacgtattg ccgccaccat ggaaaacgcc cagaaaggtg aaatcatgcc
     4621 gaacatcccg cagatgtccg ctttctggta tgccgtgcgt actgcggtga tcaacgccgc
     4681 cagcggtcgt cagactgtcg atgaagccct gaaagacgcg cagactgatt acgatatccc
     4741 aggtaccgag aacctgtact tccaatccaa tgccgcggag ctgacggtgg aggttcgcgg
     4801 ctctaacggg gctttctaca agggatttat caaagatgtt catgaagact cccttacagt
     4861 tgtttttgaa aataattggc aaccagaacg ccaggttcca tttaatgaag ttagattacc
     4921 accaccacct gatataaaaa aagaaattag tgaaggagat gaagtagagg tatattcaag
     4981 agcaaatgac caagagccat gtgggtggtg gttggctaaa gttcggatga tgaaaggaga
     5041 attttatgtc attgaatatg ctgcttgtga cgctacttac aatgaaatag tcacatttga
     5101 acgacttcgg cctgtcaatc aaaataaaac tgtcaaaaaa aataccttct ttaaatgcac
     5161 agtggatgtt cctgaggatt tgagagaggc gtgtgctaat gaaaatgcac ataaagattt
     5221 taagaaagca gtaggagcat gcagaatttt ttaccatcca gaaacaacac agctaatgat
     5281 actgtctgcc agtgaagcaa ctgtgaagag agtaaacatc ttaagtgaca tgcatttgcg
     5341 aagtattcgt acgaagttga tgcttatgtc cagaaatgaa gaggccacta agcatttaga
     5401 atgcacaaaa caacttgcag cagcttttca tgaggaattt gttgtgagag aagatttaat
     5461 gggcctggca ataggaacac atggtagtaa catccagcaa gctaggaagg ttcctggagt
     5521 taccgccatt gagctagatg aagatactgg aacattcaga atctacggag agagtgctga
     5581 tgctgtaaaa aaggctagag gtttcttgga atttgtggag gattttattc aggttcctag
     5641 gaatctcgtt ggaaaagtaa ttggaaaaaa tggcaaagtt attcaagaaa tagtggacaa
     5701 atctggtgtg gttcgagtga gaattgaagg ggacaatgaa aataaattac ccagagaaga
     5761 cggtatggtt ccatttgtat ttgttggcac taaagaaagc attggaaatg tgcaggttct
     5821 tctagagtat catattgcct atctaaagga agtagaacag ctaagaatgg aacgcctaca
     5881 gattgatgaa cagctgcgac agattggttc taggtcttat agcggaagag gcagaggtcg
     5941 tcggggacct aattacacct ccggttatgg tacaaattct gagctgtcta acccctctga
     6001 aacggaatct gagcgtaaag acgagctgag tgattggtca ttggcaggag aagatgatcg
     6061 agacagccga catcagcgtg acagcaggag acgcccagga ggaagaggca gaagtgtttc
     6121 agggggtcga ggtcgtggtg gaccacgtgg tggcaaatcc tccatcagtt ctgtgctcaa
     6181 agatccagac agcaatccat acagcttact tgataataca gaatcagatc agactgcaga
     6241 cactgatgcc agcgaatctc atcacagtac taaccgtcgt aggcggtctc gtagacgaag
     6301 gactgatgaa gatgctgttc tgatggatgg aatgactgaa tctgatacag cttcagttaa
     6361 tgaaaatggg ctaggcaaaa gatgtgatta acattggaag tggataacgg atccgaattc
     6421 gagctccgtc gacaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc
     6481 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     6541 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     6601 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     6661 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     6721 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     6781 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     6841 ggtgatggtt cacattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     6901 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     6961 accctgataa atgcttcaat aatttgaaaa aggaagagta tgagtattca acatttccgt
     7021 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     7081 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     7141 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     7201 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     7261 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     7321 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     7381 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     7441 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     7501 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     7561 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     7621 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     7681 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     7741 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     7801 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     7861 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     7921 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     7981 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aag
//
