LOCUS       sequence-212724-        7286 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 7286)
  AUTHORS   Johnson BS, Snead D, Lee JJ, McCaffery JM, Shorter J, Gitler AD
  TITLE     TDP-43 is intrinsically aggregation-prone, and amyotrophic lateral 
            sclerosis-linked mutations accelerate aggregation and increase 
            toxicity.
  JOURNAL   J Biol Chem. 2009 Jul 24. 284(30):20329-39.
  PUBMED    19465477
REFERENCE   2  (bases 1 to 7286)
  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: "J Biol 
            Chem. 2009 Jul 24. 284(30):20329-39."
COMMENT     Sequence Label: pDuet TDP43 WT
FEATURES             Location/Qualifiers
     source          1..7286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..1083)
                     /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     1045..1064
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1084..1161)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1367..1386
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1474..1492
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1493..1517
                     /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             1532..1554
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1561..2217
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MGSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
                     KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIR
                     YGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVV
                     LYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2175..2197
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2233..2253
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     promoter        3573..3591
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3592..3616
                     /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             3725..3769
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3807..3825)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3821..3868
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3905..4360
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3992..4011)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4202..4223
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(4475..5266,5267..5335))
                     /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     5098..5117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5336..5428)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5509..6097
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5998..6017
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6251..6268
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6283..6422
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6348..6367)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6508..6530
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6524..6715)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
       61 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
      121 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
      181 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
      241 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag
      301 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
      361 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
      421 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
      481 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
      541 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
      601 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
      661 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
      721 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
      781 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
      841 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
      901 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
      961 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     1021 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     1081 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     1141 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     1201 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     1261 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     1321 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     1381 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     1441 agaggatcga gatcgatctc gatcccgcga aattaatacg actcactata ggggaattgt
     1501 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc
     1561 atgggctccc ctatactagg ttattggaaa attaagggcc ttgtgcaacc cactcgactt
     1621 cttttggaat atcttgaaga aaaatatgaa gagcatttgt atgagcgcga tgaaggtgat
     1681 aaatggcgaa acaaaaagtt tgaattgggt ttggagtttc ccaatcttcc ttattatatt
     1741 gatggtgatg ttaaattaac acagtctatg gccatcatac gttatatagc tgacaagcac
     1801 aacatgttgg gtggttgtcc aaaagagcgt gcagagattt caatgcttga aggagcggtt
     1861 ttggatatta gatacggtgt ttcgagaatt gcatatagta aagactttga aactctcaaa
     1921 gttgattttc ttagcaagct acctgaaatg ctgaaaatgt tcgaagatcg tttatgtcat
     1981 aaaacatatt taaatggtga tcatgtaacc catcctgact tcatgttgta tgacgctctt
     2041 gatgttgttt tatacatgga cccaatgtgc ctggatgcgt tcccaaaatt agtttgtttt
     2101 aaaaaacgta ttgaagctat cccacaaatt gataagtact tgaaatccag caagtatata
     2161 gcatggcctt tgcagggctg gcaagccacg tttggtggtg gcgaccatcc tccaaaatcg
     2221 gatctggcta gcgagaatct ttattttcag ggatccatgt ctgaatatat tcgggtaacc
     2281 gaagatgaga acgatgagcc cattgaaata ccatcggaag acgatgggac ggtgctgctc
     2341 tccacggtta cagcccagtt tccaggggcg tgtgggcttc gctacaggaa tccagtgtct
     2401 cagtgtatga gaggtgtccg gctggtagaa ggaattctgc atgccccaga tgctggctgg
     2461 ggaaatctgg tgtatgttgt caactatcca aaagataaca aaagaaaaat ggatgagaca
     2521 gatgcttcat cagcagtgaa agtgaaaaga gcagtccaga aaacatccga tttaatagtg
     2581 ttgggtctcc catggaaaac aaccgaacag gacctgaaag agtattttag tacctttgga
     2641 gaagttctta tggtgcaggt caagaaagat cttaagactg gtcattcaaa ggggtttggc
     2701 tttgttcgtt ttacggaata tgaaacacaa gtgaaagtaa tgtcacagcg acatatgata
     2761 gatggacgat ggtgtgactg caaacttcct aattctaagc aaagccaaga tgagcctttg
     2821 agaagcagaa aagtgtttgt ggggcgctgt acagaggaca tgactgagga tgagctgcgg
     2881 gagttcttct ctcagtacgg ggatgtgatg gatgtcttca tccccaagcc attcagggcc
     2941 tttgcctttg ttacatttgc agatgatcag attgcgcagt ctctttgtgg agaggacttg
     3001 atcattaaag gaatcagcgt tcatatatcc aatgccgaac ctaagcacaa tagcaataga
     3061 cagttagaaa gaagtggaag atttggtggt aatccaggtg gctttgggaa tcagggtgga
     3121 tttggtaata gcagaggggg tggagctggt ttgggaaaca atcaaggtag taatatgggt
     3181 ggtgggatga actttggtgc gttcagcatt aatccagcca tgatggctgc cgcccaggca
     3241 gcactacaga gcagttgggg tatgatgggc atgttagcca gccagcagaa ccagtcaggc
     3301 ccatcgggta ataaccaaaa ccaaggcaac atgcagaggg agccaaacca ggccttcggt
     3361 tctggaaata actcttatag tggctctaat tctggtgcag caattggttg gggatcagca
     3421 tccaatgcag ggtcgggcag tggttttaat ggaggctttg gctcaagcat ggattctaag
     3481 tcttctggct ggggaatgta gaagcttgcg gccgcataat gcttaagtcg aacagaaagt
     3541 aatcgtattg tacacggccg cataatcgaa attaatacga ctcactatag gggaattgtg
     3601 agcggataac aattccccat cttagtatat tagttaagta taagaaggag atatacatat
     3661 ggcagatctc aattggatat cggccggcca cgcgatcgct gacgtcggta ccctcgagtc
     3721 tggtaaagaa accgctgctg cgaaatttga acgccagcac atggactcgt ctactagcgc
     3781 agcttaatta acctaggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     3841 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa
     3901 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3961 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4021 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4081 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4141 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4201 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4261 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4321 tttaacgcga attttaacaa aatattaacg tttacaattt ctggcggcac gatggcatga
     4381 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4441 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4501 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4561 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4621 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4681 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4741 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4801 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4861 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4921 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4981 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5041 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5101 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5161 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5221 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5281 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5341 tcctttttca atcatgattg aagcatttat cagggttatt gtctcatgag cggatacata
     5401 tttgaatgta tttagaaaaa taaacaaata ggtcatgacc aaaatccctt aacgtgagtt
     5461 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5521 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5581 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5641 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5701 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5761 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5821 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5881 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5941 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6001 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6061 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6121 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6181 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6241 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     6301 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     6361 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     6421 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     6481 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     6541 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     6601 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     6661 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     6721 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     6781 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     6841 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     6901 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     6961 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     7021 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     7081 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     7141 gccgggtcct caacgacagg agcacgatca tgctagtcat gccccgcgcc caccggaagg
     7201 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     7261 gctaacttac attaattgcg ttgcgc
//
