LOCUS       sequence-430099-        4602 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses 6xHis-tagged C-terminal prion-like domains of Efg1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4602)
  AUTHORS   Wang SH, Zheng T, Fawzi NL
  TITLE     Structure and position-specific interactions of prion-like domains 
            in transcription factor Efg1 phase separation.
  JOURNAL   bioRxiv [Preprint]. 2023 Nov 9:2023.11.09.566450. doi: 
            10.1101/2023.11.09.566450.
  PUBMED    37986834
REFERENCE   2  (bases 1 to 4602)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2023 Nov 9:2023.11.09.566450. doi: 
            10.1101/2023.11.09.566450."
COMMENT     Sequence Label: Efg1-C-PrLD
FEATURES             Location/Qualifiers
     source          1..4602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..21
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     255..272
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     370..387
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     371..387
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     851..870
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        851..869
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    870..894
                     /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             955..972
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             973..993
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      1607..1654
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1877..1896)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        1963..2054
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2055..2864
                     /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             2947..4029
                     /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(2966..2985)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      join(4115..4602,1..101)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
       61 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
      121 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
      181 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
      241 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga aggcgagagt agggaactgc
      301 caggcatcaa actaagcaga aggcccctga cggatggcct ttttgcgttt ctacaaactc
      361 tttctgtgtt gtaaaacgac ggccagtctt aagctcgggc cccctgggcg gttctgataa
      421 cgagtaatcg ttaatccgca aataacgtaa aaacccgctt cggcgggttt ttttatgggg
      481 ggagtttagg gaaagagcat ttgtcagaat atttaagggc gcctgtcact ttgcttgata
      541 tatgagaatt atttaacctt ataaatgaga aaaaagcaac gcactttaaa taagatacgt
      601 tgctttttcg attgatgaac acctataatt aaactattca tctattattt atgatttttt
      661 gtatatacaa tatttctagt ttgttaaaga gaattaagaa aataaatctc gaaaataata
      721 aagggaaaat cagtttttga tatcaaaatt atacatgtca acgataatac aaaatataat
      781 acaaactata agatgttatc agtatttatt atgcatttag aataaatttt gtgtcgccct
      841 tccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta
      901 gaaataattt tgtttaactt ttgaaggaga tataccatgg gcagcgacaa gatccaccat
      961 caccaccatc acgaaaatct gtactttcaa ggtcatatgt caggtaatgg taatagcatt
     1021 agcgcgacca gcggtggtag caatgttagt ggtgcaagcg gtgcaggtag caccaccagt
     1081 ccggttaata ccaaagcagc gaccgcagcc ggtattccgc agggcaacta ttatcagacc
     1141 tataatcagc aacagtaccc gcagcagtat ggtcagtata atgcaccggg taaaaatcag
     1201 aatacaccgg catcacagcc tggtagtacc accaatgatc agtatctgca gcagcagcaa
     1261 cagcaacaac aacaaatgta tggttatcag agcaactact atcaaggtgg tgcagccaat
     1321 agcagctatt atccgaatta ttaccaacaa cagcagccga attatgcaag cagctatccg
     1381 tatcaacagc agcaacaaaa acagcagcaa caacaaccga atcagcagca acagagtgat
     1441 cagcagcaga cctcaacccc gagcggtggt gccggtacac gtagcgttca tcagagtccg
     1501 caggttcaga gcctgaccca gggtagcgtg catccgagcc cacagcagca tcaggcaaat
     1561 cagagcgcaa gcaccgttgc aaaagaagag aaataactcg agccccctag cataacccct
     1621 tggggcctct aaacgggtct tgaggggttt tttgcccctg agacgcgtca atcgagttcg
     1681 tacctaaggg cgacaccccc taattagccc gggcgaaagg cccagtcttt cgactgagcc
     1741 tttcgtttta tttgatgcct ggcagttccc tactctcgca tggggagtcc ccacactacc
     1801 atcggcgcta cggcgtttca cttctgagtt cggcatgggg tcaggtggga ccaccgcgct
     1861 actgccgcca ggcaaacaag gggtgttatg agccatattc aggtataaat gggctcgcga
     1921 taatgttcag aattggttaa ttggttgtaa cactgacccc tatttgttta tttttctaaa
     1981 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2041 gaaaaaggaa gaatatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca
     2101 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     2161 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     2221 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     2281 ttatgccact tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     2341 ccactgcgat ccccggaaaa acagcgttcc aggtattaga agaatatcct gattcaggtg
     2401 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcactcgatt cctgtttgta
     2461 attgtccttt taacagcgat cgcgtatttc gcctcgctca ggcgcaatca cgaatgaata
     2521 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     2581 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     2641 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     2701 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     2761 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     2821 tgaataaatt gcaatttcat ttgatgctcg atgagttttt ctaagcggcg cgccatcgaa
     2881 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     2941 gtgaatatga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     3001 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     3061 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
     3121 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
     3181 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
     3241 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
     3301 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
     3361 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
     3421 attattttct cccatgagga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
     3481 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
     3541 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc
     3601 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt
     3661 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc
     3721 gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagat
     3781 agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa
     3841 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg
     3901 ttgccagtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct
     3961 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     4021 gggcagtgac tcatgaccaa aatcccttaa cgtgagttac gcgcgcgtcg ttccactgag
     4081 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4141 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4201 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4261 ttcttctagt gtagccgtag ttagcccacc acttcaagaa ctctgtagca ccgcctacat
     4321 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4381 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4441 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4501 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4561 gcggcagggt cggaacagga gagcgcacga gggagcttcc ag
//
