LOCUS       sequence-189261-        5823 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses untagged FUS AA 1-163.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5823)
  AUTHORS   Burke KA, Janke AM, Rhine CL, Fawzi NL
  TITLE     Residue-by-Residue View of In Vitro FUS Granules that Bind the 
            C-Terminal Domain of RNA Polymerase II.
  JOURNAL   Mol Cell. 2015 Oct 15;60(2):231-41. doi: 
            10.1016/j.molcel.2015.09.006. Epub 2015 Oct 8.
  PUBMED    26455390
REFERENCE   2  (bases 1 to 5823)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.molcel.2015.09.006"; journalName: "Mol Cell"; date: 
            "2015-10-15- 15"; volume: "60"; issue: "2"; pages: "231-41"
COMMENT     Sequence Label: FUS_anion
FEATURES             Location/Qualifiers
     source          1..5823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             60..80
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             464..475
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(625..643)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      639..686
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      723..1178
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(810..829)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1020..1041
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1204..1308
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1309..1377,1378..2169)
                     /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(1527..1546)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2340..2928
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2829..2848
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3082..3099
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3114..3253
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3179..3198)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3339..3361
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3355..3546)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4355..5437)
                     /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     5399..5418
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5438..5515)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5721..5740
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag catatggcct caaacgatta tacccaacaa gcaacccaaa
      121 gctatggggc ctaccccacc cagcccgggc agggctattc ccagcagagc agtcagccct
      181 acggacagca gagttacagt ggttatagcc agtccacgga cacttcaggc tatggccaga
      241 gcagctattc ttcttatggc cagagccaga acacaggcta tggaactcag tcaactcccc
      301 agggatatgg ctcgactggc ggctatggca gtagccagag ctcccaatcg tcttacgggc
      361 agcagtcctc ctatcctggc tatggccagc agccagctcc cagcagcacc tcgggaagtt
      421 acggtagcag ttctcagagc agcagctatg ggcagcccca gagtgggagc tacagccagc
      481 agcctagcta tggtggacag cagcaaagct atggacagca gcaaagctat aatccccctc
      541 agggctatgg acagcagaac cagtacaaca gctgagatcc ggctgctaac aaagcccgaa
      601 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
      661 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg
      721 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
      781 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
      841 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
      901 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
      961 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     1021 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     1081 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     1141 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     1201 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     1261 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     1321 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     1381 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1441 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     1501 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     1561 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1621 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     1681 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1741 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1801 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     1861 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1921 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1981 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     2041 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     2101 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     2161 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     2221 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     2281 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2341 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2401 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2461 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2521 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2581 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2641 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2701 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2761 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2821 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2881 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2941 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3001 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3061 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     3121 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     3181 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     3241 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3301 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3361 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     3421 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     3481 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     3541 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     3601 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     3661 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     3721 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     3781 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     3841 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     3901 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     3961 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     4021 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     4081 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     4141 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     4201 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     4261 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     4321 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4381 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4441 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     4501 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     4561 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     4621 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     4681 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     4741 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     4801 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     4861 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     4921 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     4981 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     5041 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     5101 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     5161 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     5221 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     5281 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     5341 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     5401 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     5461 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     5521 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     5581 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     5641 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     5701 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     5761 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
     5821 aat
//
