LOCUS       sequence-430100-        4533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged DBD of Efg1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4533)
  AUTHORS   Wang SH, Zheng T, Fawzi NL
  TITLE     Structure and interactions of prion-like domains in transcription 
            factor Efg1 phase separation.
  JOURNAL   Biophys J. 2024 Jun 4;123(11):1481-1493. doi: 
            10.1016/j.bpj.2024.01.030. Epub 2024 Feb 1.
  PUBMED    38297837
REFERENCE   2  (bases 1 to 4533)
  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"; doi: 
            "10.1016/j.bpj.2024.01.030"; journalName: "Biophys J"; date: 
            "2024-06-4- 4"; volume: "123"; issue: "11"; pages: "1481-1493"
COMMENT     Sequence Label: Efg1-DBD
FEATURES             Location/Qualifiers
     source          1..4533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     130..149
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     383..400
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     498..515
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     499..515
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     979..998
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        979..997
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    998..1022
                     /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             1083..1100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1101..1121
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      1666..1713
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1936..1955)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2022..2113
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2114..2923
                     /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             3006..4088
                     /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(3025..3044)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      join(4174..4533,1..229)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
       61 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      121 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      181 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      241 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      301 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      361 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag gcgagagtag
      421 ggaactgcca ggcatcaaac taagcagaag gcccctgacg gatggccttt ttgcgtttct
      481 acaaactctt tctgtgttgt aaaacgacgg ccagtcttaa gctcgggccc cctgggcggt
      541 tctgataacg agtaatcgtt aatccgcaaa taacgtaaaa acccgcttcg gcgggttttt
      601 ttatgggggg agtttaggga aagagcattt gtcagaatat ttaagggcgc ctgtcacttt
      661 gcttgatata tgagaattat ttaaccttat aaatgagaaa aaagcaacgc actttaaata
      721 agatacgttg ctttttcgat tgatgaacac ctataattaa actattcatc tattatttat
      781 gattttttgt atatacaata tttctagttt gttaaagaga attaagaaaa taaatctcga
      841 aaataataaa gggaaaatca gtttttgata tcaaaattat acatgtcaac gataatacaa
      901 aatataatac aaactataag atgttatcag tatttattat gcatttagaa taaattttgt
      961 gtcgcccttc cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt
     1021 cccctctaga aataattttg tttaactttt gaaggagata taccatgggc agcgacaaga
     1081 tccaccatca ccaccatcac gaaaatctgt actttcaagg tcatatgccg gttcaggata
     1141 ccctgaatgc cagcagcacc agtaccgttg gtcagtttca gcctccgggt attcgtccgc
     1201 gtgttaccac caccatgtgg gaagatgaaa aaaccctgtg ttatcaggtg gatgccaata
     1261 atgttagcgt tgttcgtcgt gcagataaca acatgattaa tggcaccaaa ctgctgaatg
     1321 ttgcacagat gacccgtggt cgtcgtgatg gtattctgaa aagcgaaaaa gttcgtcacg
     1381 tggtgaaaat tggtagcatg catctgaaag gtgtttggat tccgtttgaa cgtgcactgg
     1441 caatggcaca gcgtgagcag attgttgata tgctgtatcc gctgtttgtg cgtgatatta
     1501 aacgtgttat tcagaccggt gttaccccga atgcggcagc ggctactgca gctgccgcag
     1561 ccaccgcaac cagcgcaagt gcacctcctc cgcctccgcc accggttgca gcagctacca
     1621 caacagcggc aaccgccatt agcaaaagca gctaactcga gccccctagc ataacccctt
     1681 ggggcctcta aacgggtctt gaggggtttt ttgcccctga gacgcgtcaa tcgagttcgt
     1741 acctaagggc gacaccccct aattagcccg ggcgaaaggc ccagtctttc gactgagcct
     1801 ttcgttttat ttgatgcctg gcagttccct actctcgcat ggggagtccc cacactacca
     1861 tcggcgctac ggcgtttcac ttctgagttc ggcatggggt caggtgggac caccgcgcta
     1921 ctgccgccag gcaaacaagg ggtgttatga gccatattca ggtataaatg ggctcgcgat
     1981 aatgttcaga attggttaat tggttgtaac actgacccct atttgtttat ttttctaaat
     2041 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2101 aaaaaggaag aatatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     2161 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     2221 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     2281 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     2341 tatgccactt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     2401 cactgcgatc cccggaaaaa cagcgttcca ggtattagaa gaatatcctg attcaggtga
     2461 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cactcgattc ctgtttgtaa
     2521 ttgtcctttt aacagcgatc gcgtatttcg cctcgctcag gcgcaatcac gaatgaataa
     2581 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     2641 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     2701 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     2761 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     2821 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     2881 gaataaattg caatttcatt tgatgctcga tgagtttttc taagcggcgc gccatcgaat
     2941 ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
     3001 tgaatatgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga
     3061 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg
     3121 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca
     3181 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa
     3241 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg
     3301 tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg
     3361 tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg
     3421 cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta
     3481 ttattttctc ccatgaggac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc
     3541 accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg
     3601 ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg
     3661 actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc
     3721 ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg
     3781 agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagata
     3841 gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa
     3901 ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt
     3961 tgccagtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc
     4021 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     4081 ggcagtgact catgaccaaa atcccttaac gtgagttacg cgcgcgtcgt tccactgagc
     4141 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4201 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4261 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4321 tcttctagtg tagccgtagt tagcccacca cttcaagaac tctgtagcac cgcctacata
     4381 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4441 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4501 ttcgtgcaca cagcccagct tggagcgaac gac
//
