LOCUS       sequence-189299-        4260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged C-terminal domain of RNA polymerase II.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4260)
  AUTHORS   Janke AM, Seo DH, Rahmanian V, Conicella AE, Mathews KL, Burke KA, 
            Mittal J, Fawzi NL
  TITLE     Lysines in the RNA Polymerase II C-Terminal Domain Contribute to 
            TAF15 Fibril Recruitment.
  JOURNAL   Biochemistry. 2017 Oct 11. doi: 10.1021/acs.biochem.7b00310.
  PUBMED    28945358
REFERENCE   2  (bases 1 to 4260)
  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: 
            "Biochemistry. 2017 Oct 11. doi: 10.1021/acs.biochem.7b00310."
COMMENT     Sequence Label: RNAPII_CTD43-52
FEATURES             Location/Qualifiers
     source          1..4260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             105..122
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             123..143
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      415..462
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(685..704)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        771..862
                     /gene="bla"
                     /label=AmpR promoter
     CDS             863..1672
                     /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             1755..2837
                     /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(1774..1793)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      2923..3511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3412..3431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3665..3682
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3780..3797
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3781..3797
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt ttgaaggaga tataccatgg gcagcgataa gatccaccac caccatcatc
      121 atgagaatct gtattttcaa ggtcatatgt acagcccaac gtcgccgacg tattcgccta
      181 ccagcccggt ttacaccccg acctctccga aatactcccc tacgagcccg acttactctc
      241 cgactagccc gaagtactct ccaacctccc cgacctacag ccctaccagc ccaaagggca
      301 gcacctattc accgacgagc ccgggttact cgccgaccag ccctacctac agcctgacca
      361 gcccagcgat tagcccggac gatagcgacg aagagaacta ataactcgag ccccctagca
      421 taaccccttg gggcctctaa acgggtcttg aggggttttt tgcccctgag acgcgtcaat
      481 cgagttcgta cctaagggcg acacccccta attagcccgg gcgaaaggcc cagtctttcg
      541 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcatg gggagtcccc
      601 acactaccat cggcgctacg gcgtttcact tctgagttcg gcatggggtc aggtgggacc
      661 accgcgctac tgccgccagg caaacaaggg gtgttatgag ccatattcag gtataaatgg
      721 gctcgcgata atgttcagaa ttggttaatt ggttgtaaca ctgaccccta tttgtttatt
      781 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      841 ataatattga aaaaggaaga atatgagcca tattcaacgg gaaacgtcga ggccgcgatt
      901 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
      961 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     1021 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     1081 gacggaattt atgccacttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     1141 gttactcacc actgcgatcc ccggaaaaac agcgttccag gtattagaag aatatcctga
     1201 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc actcgattcc
     1261 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgc ctcgctcagg cgcaatcacg
     1321 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     1381 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     1441 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     1501 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     1561 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     1621 tcctgatatg aataaattgc aatttcattt gatgctcgat gagtttttct aagcggcgcg
     1681 ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
     1741 tcagggtggt gaatatgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     1801 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     1861 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     1921 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     1981 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     2041 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     2101 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     2161 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     2221 tcaacagtat tattttctcc catgaggacg gtacgcgact gggcgtggag catctggtcg
     2281 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     2341 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     2401 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     2461 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     2521 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
     2581 ccgaagatag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
     2641 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     2701 atcagctgtt gccagtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     2761 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     2821 tggaaagcgg gcagtgactc atgaccaaaa tcccttaacg tgagttacgc gcgcgtcgtt
     2881 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2941 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3001 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3061 aaatactgtt cttctagtgt agccgtagtt agcccaccac ttcaagaact ctgtagcacc
     3121 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3181 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3241 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3301 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3361 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3421 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3481 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3541 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3601 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3661 gcgcagcgag tcagtgagcg aggaagcgga aggcgagagt agggaactgc caggcatcaa
     3721 actaagcaga aggcccctga cggatggcct ttttgcgttt ctacaaactc tttctgtgtt
     3781 gtaaaacgac ggccagtctt aagctcgggc cccctgggcg gttctgataa cgagtaatcg
     3841 ttaatccgca aataacgtaa aaacccgctt cggcgggttt ttttatgggg ggagtttagg
     3901 gaaagagcat ttgtcagaat atttaagggc gcctgtcact ttgcttgata tatgagaatt
     3961 atttaacctt ataaatgaga aaaaagcaac gcactttaaa taagatacgt tgctttttcg
     4021 attgatgaac acctataatt aaactattca tctattattt atgatttttt gtatatacaa
     4081 tatttctagt ttgttaaaga gaattaagaa aataaatctc gaaaataata aagggaaaat
     4141 cagtttttga tatcaaaatt atacatgtca acgataatac aaaatataat acaaactata
     4201 agatgttatc agtatttatt atgcatttag aataaatttt gtgtcgccct tccgcgaaat
//
