LOCUS       sequence-381059-        4452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged C-terminal domain of TDP-43 with 5M-->A_CR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4452)
  AUTHORS   Mohanty P, Shenoy J, Rizuan A, Mercado-Ortiz JF, Fawzi NL, Mittal J
  TITLE     A synergy between site-specific and transient interactions drives 
            the phase separation of a disordered, low-complexity domain.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Aug 22;120(34):e2305625120. doi: 
            10.1073/pnas.2305625120. Epub 2023 Aug 14.
  PUBMED    37579155
REFERENCE   2  (bases 1 to 4452)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2023-08-22"; volume: "120(34):e2305625120. 
            doi"; pages: " 10.1073/pnas.2305625120. Epub 2023 Aug 14"
COMMENT     Sequence Label: TDP-43 _CTD_5M-->A_CR
FEATURES             Location/Qualifiers
     source          1..4452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      166..754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     655..674
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     908..925
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1023..1040
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1024..1040
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1504..1523
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1504..1522
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1523..1547
                     /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             1608..1625
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1626..1646
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      2110..2157
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2380..2399)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2466..2557
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2558..3367
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             join(3450..4452,1..80)
                     /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     complement(3469..3488)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
       61 gactggaaag cgggcagtga ctcatgacca aaatccctta acgtgagtta cgcgcgcgtc
      121 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
      181 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
      241 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
      301 accaaatact gttcttctag tgtagccgta gttagcccac cacttcaaga actctgtagc
      361 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
      421 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
      481 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
      541 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
      601 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
      661 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
      721 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
      781 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
      841 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
      901 cgagcgcagc gagtcagtga gcgaggaagc ggaaggcgag agtagggaac tgccaggcat
      961 caaactaagc agaaggcccc tgacggatgg cctttttgcg tttctacaaa ctctttctgt
     1021 gttgtaaaac gacggccagt cttaagctcg ggccccctgg gcggttctga taacgagtaa
     1081 tcgttaatcc gcaaataacg taaaaacccg cttcggcggg tttttttatg gggggagttt
     1141 agggaaagag catttgtcag aatatttaag ggcgcctgtc actttgcttg atatatgaga
     1201 attatttaac cttataaatg agaaaaaagc aacgcacttt aaataagata cgttgctttt
     1261 tcgattgatg aacacctata attaaactat tcatctatta tttatgattt tttgtatata
     1321 caatatttct agtttgttaa agagaattaa gaaaataaat ctcgaaaata ataaagggaa
     1381 aatcagtttt tgatatcaaa attatacatg tcaacgataa tacaaaatat aatacaaact
     1441 ataagatgtt atcagtattt attatgcatt tagaataaat tttgtgtcgc ccttccgcga
     1501 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
     1561 ttttgtttaa cttttgaagg agatatacca tgggcagcga caagatccac catcaccacc
     1621 atcacgaaaa tctgtacttt caaggtcata tgaaccgtca gctggagcgt agcggtcgtt
     1681 tcggtggcaa cccgggtggc ttcggcaacc aaggtggctt tggtaacagc cgtggtggcg
     1741 gtgcgggtct gggtaacaac cagggtagca acatgggcgg tggcatgaac ttcggtgcgt
     1801 ttagcatcaa cccggcggcg gcggcggcgg cgcaggcggc gctgcaaagc agctggggcg
     1861 cggcgggtgc gctggcgagc cagcaaaacc aaagcggccc gagcggtaac aaccagaacc
     1921 aaggcaacat gcagcgtgaa ccgaaccaag cgtttggtag cggcaacaac agctacagcg
     1981 gcagcaacag cggtgcggcg attggttggg gcagcgcgag caacgcgggt agcggtagcg
     2041 gttttaacgg tggctttggt agcagcatgg acagcaagag cagcggctgg ggcatgtaac
     2101 tcgagccccc tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgccc
     2161 ctgagacgcg tcaatcgagt tcgtacctaa gggcgacacc ccctaattag cccgggcgaa
     2221 aggcccagtc tttcgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc
     2281 gcatggggag tccccacact accatcggcg ctacggcgtt tcacttctga gttcggcatg
     2341 gggtcaggtg ggaccaccgc gctactgccg ccaggcaaac aaggggtgtt atgagccata
     2401 ttcaggtata aatgggctcg cgataatgtt cagaattggt taattggttg taacactgac
     2461 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2521 ctgataaatg cttcaataat attgaaaaag gaagaatatg agccatattc aacgggaaac
     2581 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2641 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     2701 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2761 cagactaaac tggctgacgg aatttatgcc acttccgacc atcaagcatt ttatccgtac
     2821 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcgt tccaggtatt
     2881 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2941 gttgcactcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgcctcgc
     3001 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3061 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     3121 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3181 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     3241 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3301 atatggtatt gataatcctg atatgaataa attgcaattt catttgatgc tcgatgagtt
     3361 tttctaagcg gcgcgccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc
     3421 cggaagagag tcaattcagg gtggtgaata tgaaaccagt aacgttatac gatgtcgcag
     3481 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt
     3541 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc
     3601 gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc
     3661 tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg
     3721 gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg
     3781 tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc
     3841 aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct
     3901 ctgaccagac acccatcaac agtattattt tctcccatga ggacggtacg cgactgggcg
     3961 tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt
     4021 ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc
     4081 agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc
     4141 aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc
     4201 tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag
     4261 tgggatacga cgataccgaa gatagctcat gttatatccc gccgttaacc accatcaaac
     4321 aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc
     4381 aggcggtgaa gggcaatcag ctgttgccag tctcactggt gaaaagaaaa accaccctgg
     4441 cgcccaatac gc
//
