LOCUS       sequence-241907-        7721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7721)
  AUTHORS   Sun Z, Diaz Z, Fang X, Hart MP, Chesi A, Shorter J, Gitler AD
  TITLE     Molecular determinants and genetic modifiers of aggregation and 
            toxicity for the ALS disease protein FUS/TLS.
  JOURNAL   PLoS Biol. 2011 Apr;9(4):e1000614. doi: 
            10.1371/journal.pbio.1000614. Epub 2011 Apr 26.
  PUBMED    21541367
REFERENCE   2  (bases 1 to 7721)
  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: "PLoS 
            Biol."; date: "2011-04"; volume: "9(4):e1000614. doi"; pages: " 
            10.1371/journal.pbio.1000614. Epub 2011 Apr 26"
COMMENT     Sequence Label: GST-TEV-FUS
FEATURES             Location/Qualifiers
     source          1..7721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     246..265
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        353..371
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    372..396
                     /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             411..433
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             440..1096
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MGSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
                     KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIR
                     YGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVV
                     LYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1054..1076
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1112..1132
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    1139..1163
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1549..1560
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(2751..2775)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        2887..2905
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2906..2930
                     /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)."
     CDS             3039..3083
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3121..3139)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3135..3182
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3219..3674
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3306..3325)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3516..3537
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(3789..4580,4581..4649))
                     /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     4412..4431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4650..4742)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4823..5411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5312..5331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5565..5582
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5597..5736
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5662..5681)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5822..5844
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5838..6029)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6601..7683)
                     /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     7645..7664
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(join(7684..7721,1..40))
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
       61 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
      121 gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
      181 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
      241 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
      301 gccacgatgc gtccggcgta gaggatcgag atcgatctcg atcccgcgaa attaatacga
      361 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
      421 tttaagaagg agatatacca tgggctcccc tatactaggt tattggaaaa ttaagggcct
      481 tgtgcaaccc actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta
      541 tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc
      601 caatcttcct tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg
      661 ttatatagct gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc
      721 aatgcttgaa ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa
      781 agactttgaa actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt
      841 cgaagatcgt ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt
      901 catgttgtat gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt
      961 cccaaaatta gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt
     1021 gaaatccagc aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg
     1081 cgaccatcct ccaaaatcgg atctggctag cgagaatctt tattttcagg gatcatcaac
     1141 aagtttgtac aaaaaagcag gctccaccat ggcctcaaac gattataccc aacaagcaac
     1201 ccaaagctat ggggcctacc ccacccagcc cgggcagggc tattcccagc agagcagtca
     1261 gccctacgga cagcagagtt acagtggtta tagccagtcc acggacactt caggctatgg
     1321 ccagagcagc tattcttctt atggccagag ccagaacaca ggctatggaa ctcagtcaac
     1381 tccccaggga tatggctcga ctggcggcta tggcagtagc cagagctccc aatcgtctta
     1441 cgggcagcag tcctcctatc ctggctatgg ccagcagcca gctcccagca gcacctcggg
     1501 aagttacggt agcagttctc agagcagcag ctatgggcag ccccagagtg ggagctacag
     1561 ccagcagcct agctatggtg gacagcagca aagctatgga cagcagcaaa gctataatcc
     1621 ccctcagggc tatggacagc agaaccagta caacagcagc agtggtggtg gaggtggagg
     1681 tggaggtgga ggtaactatg gccaagatca atcctccatg agtagtggtg gtggcagtgg
     1741 tggcggttat ggcaatcaag accagagtgg tggaggtggc agcggtggct atggacagca
     1801 ggaccgtgga ggccgcggca ggggtggcag tggtggcggc ggcggcggcg gcggtggtgg
     1861 ttacaaccgc agcagtggtg gctatgaacc cagaggtcgt ggaggtggcc gtggaggcag
     1921 aggtggcatg ggcggaagtg accgtggtgg cttcaataaa tttggtggcc ctcgggacca
     1981 aggatcacgt catgactccg aacaggataa ttcagacaac aacaccatct ttgtgcaagg
     2041 cctgggtgag aatgttacaa ttgagtctgt ggctgattac ttcaagcaga ttggtattat
     2101 taagacaaac aagaaaacgg gacagcccat gattaatttg tacacagaca gggaaactgg
     2161 caagctgaag ggagaggcaa cggtctcttt tgatgaccca ccttcagcta aagcagctat
     2221 tgactggttt gatggtaaag aattctccgg aaatcctatc aaggtctcat ttgctactcg
     2281 ccgggcagac tttaatcggg gtggtggcaa tggtcgtgga ggccgagggc gaggaggacc
     2341 catgggccgt ggaggctatg gaggtggtgg cagtggtggt ggtggccgag gaggatttcc
     2401 cagtggaggt ggtggcggtg gaggacagca gcgagctggt gactggaagt gtcctaatcc
     2461 cacctgtgag aatatgaact tctcttggag gaatgaatgc aaccagtgta aggcccctaa
     2521 accagatggc ccaggagggg gaccaggtgg ctctcacatg gggggtaact acggggatga
     2581 tcgtcgtggt ggcagaggag gctatgatcg aggcggctac cggggccgcg gcggggaccg
     2641 tggaggcttc cgagggggcc ggggtggtgg ggacagaggt ggctttggcc ctggcaagat
     2701 ggattccagg ggtgagcaca gacaggatcg cagggagagg ccgtattagg acccagcttt
     2761 cttgtacaaa gtggttgatg atccgaattc gagctcggcg cgcctgcagg tcgacaagct
     2821 tgcggccgca taatgcttaa gtcgaacaga aagtaatcgt attgtacacg gccgcataat
     2881 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc ccatcttagt
     2941 atattagtta agtataagaa ggagatatac atatggcaga tctcaattgg atatcggccg
     3001 gccacgcgat cgctgacgtc ggtaccctcg agtctggtaa agaaaccgct gctgcgaaat
     3061 ttgaacgcca gcacatggac tcgtctacta gcgcagctta attaacctag gctgctgcca
     3121 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     3181 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt
     3241 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3301 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3361 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3421 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3481 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3541 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3601 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3661 aacgtttaca atttctggcg gcacgatggc atgagattat caaaaaggat cttcacctag
     3721 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3781 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3841 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3901 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3961 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4021 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4081 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4141 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4201 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4261 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4321 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4381 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4441 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4501 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4561 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4621 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatcatg attgaagcat
     4681 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4741 aataggtcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4801 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4861 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4921 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     4981 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5041 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5101 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5161 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5221 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5281 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5341 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5401 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5461 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5521 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5581 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     5641 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     5701 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     5761 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5821 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     5881 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     5941 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     6001 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     6061 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     6121 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     6181 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
     6241 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     6301 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
     6361 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     6421 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
     6481 atcatgctag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     6541 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     6601 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     6661 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     6721 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     6781 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     6841 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag
     6901 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     6961 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     7021 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     7081 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     7141 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     7201 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     7261 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     7321 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     7381 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     7441 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     7501 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     7561 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     7621 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     7681 cacattcacc accctgaatt gactctcttc cgggcgctat c
//
