LOCUS       sequence-189295-        4605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged C-terminal domain of RNA polymerase II with 
            N2A and S1966C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4605)
  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 4605)
  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_CTD27-52_N2A_S1966C
FEATURES             Location/Qualifiers
     source          1..4605
                     /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      760..807
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1030..1049)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        1116..1207
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1208..2017
                     /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             2100..3182
                     /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(2119..2138)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      3268..3856
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3757..3776
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4010..4027
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4125..4142
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4126..4142
                     /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 ggtcatatga gcccggctta cacgccgacc tcgccggcct
      181 attctcctac ctccccgagc tatagcccta ctagcccgag ctactccccg accagcccaa
      241 gctacagccc gagctccccg cgttacacgc cgcagagccc gacctatacg ccgagctctc
      301 cgagctattc cccgagcagc ccgtcttata gcccggcgag cccgaaatat accccaacct
      361 ctccgtctta cagcccgtcc agcccggaat acactccgac gtccccgaaa tacagcccga
      421 cttcgccgaa atattccccg acctccccga agtatagccc aaccagcccg acgtacagcc
      481 cgaccacccc gaagtacagc ccaacgtcgc cgacgtattc gcctaccagc ccggtttaca
      541 ccccgacctc tccgaaatac tcccctacga gcccgactta ctctccgact agcccgaagt
      601 actctccaac ctccccgacc tacagcccta ccagcccaaa gggcagcacc tattcaccga
      661 cgagcccggg ttactcgccg accagcccta cctacagcct gaccagccca gcgattagcc
      721 cggacgattg cgacgaagag aactaataac tcgagccccc tagcataacc ccttggggcc
      781 tctaaacggg tcttgagggg ttttttgccc ctgagacgcg tcaatcgagt tcgtacctaa
      841 gggcgacacc ccctaattag cccgggcgaa aggcccagtc tttcgactga gcctttcgtt
      901 ttatttgatg cctggcagtt ccctactctc gcatggggag tccccacact accatcggcg
      961 ctacggcgtt tcacttctga gttcggcatg gggtcaggtg ggaccaccgc gctactgccg
     1021 ccaggcaaac aaggggtgtt atgagccata ttcaggtata aatgggctcg cgataatgtt
     1081 cagaattggt taattggttg taacactgac ccctatttgt ttatttttct aaatacattc
     1141 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1201 gaagaatatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     1261 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     1321 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     1381 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     1441 acttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     1501 gatccccgga aaaacagcgt tccaggtatt agaagaatat cctgattcag gtgaaaatat
     1561 tgttgatgcg ctggcagtgt tcctgcgccg gttgcactcg attcctgttt gtaattgtcc
     1621 ttttaacagc gatcgcgtat ttcgcctcgc tcaggcgcaa tcacgaatga ataacggttt
     1681 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     1741 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     1801 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     1861 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     1921 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     1981 attgcaattt catttgatgc tcgatgagtt tttctaagcg gcgcgccatc gaatggcgca
     2041 aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaata
     2101 tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt
     2161 cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg
     2221 cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt
     2281 cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg
     2341 cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac
     2401 gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg
     2461 ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca
     2521 ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt
     2581 tctcccatga ggacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc
     2641 aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct
     2701 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
     2761 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
     2821 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
     2881 ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa gatagctcat
     2941 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
     3001 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgccag
     3061 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
     3121 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     3181 gactcatgac caaaatccct taacgtgagt tacgcgcgcg tcgttccact gagcgtcaga
     3241 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3301 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3361 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3421 agtgtagccg tagttagccc accacttcaa gaactctgta gcaccgccta catacctcgc
     3481 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3541 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3601 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3661 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3721 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3781 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3841 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3901 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3961 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     4021 gagcgaggaa gcggaaggcg agagtaggga actgccaggc atcaaactaa gcagaaggcc
     4081 cctgacggat ggcctttttg cgtttctaca aactctttct gtgttgtaaa acgacggcca
     4141 gtcttaagct cgggccccct gggcggttct gataacgagt aatcgttaat ccgcaaataa
     4201 cgtaaaaacc cgcttcggcg ggttttttta tggggggagt ttagggaaag agcatttgtc
     4261 agaatattta agggcgcctg tcactttgct tgatatatga gaattattta accttataaa
     4321 tgagaaaaaa gcaacgcact ttaaataaga tacgttgctt tttcgattga tgaacaccta
     4381 taattaaact attcatctat tatttatgat tttttgtata tacaatattt ctagtttgtt
     4441 aaagagaatt aagaaaataa atctcgaaaa taataaaggg aaaatcagtt tttgatatca
     4501 aaattataca tgtcaacgat aatacaaaat ataatacaaa ctataagatg ttatcagtat
     4561 ttattatgca tttagaataa attttgtgtc gcccttccgc gaaat
//
