LOCUS       sequence-290177-        6845 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsNot1iso1_1565-2371.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6845)
  AUTHORS   Boland A, Chen Y, Raisch T, Jonas S, Kuzuoglu-Ozturk D, Wohlbold L, 
            Weichenrieder O, Izaurralde E
  TITLE     Structure and assembly of the NOT module of the human CCR4-NOT 
            complex.
  JOURNAL   Nat Struct Mol Biol. 2013 Nov;20(11):1289-97. doi: 
            10.1038/nsmb.2681. Epub 2013 Oct 13.
  PUBMED    24121232
REFERENCE   2  (bases 1 to 6845)
  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: "10.1038/nsmb.2681";
            journalName: "Nat Struct Mol Biol"; date: "2013-11"; volume: "20"; 
            issue: "11"; pages: "1289-97"
COMMENT     Sequence Label: pnYC-NvH-HsNot1iso1_1565-2371_U
FEATURES             Location/Qualifiers
     source          1..6845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2294..2313
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2402..2421
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2402..2420
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2421..2445
                     /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)."
     RBS             2460..2482
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2501..2518
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2540..2560
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(5077..5095)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5091..5138
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5313..5318,5319..5335,5336..5341))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      5453..5998
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     5899..5918
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6115..6132
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        6524..6626
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(6627..6845,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6693..6712)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ggtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
       61 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
      121 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
      181 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
      241 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
      301 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
      361 gccgtctgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
      421 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt
      481 gctacgcctg aataagtgat aataagcgga tgaatggcag aaattcgaaa gcaaattcga
      541 cccggtcgtc ggttcagggc agggtcgtta aatagccgct tatgtctatt gctggtttac
      601 cggtttattg actaccggaa gcagtgtgac cgtgtgcttc tcaaatgcct gaggccagtt
      661 tgctcaggct ctccccgtgg aggtaataat tgacgatatg atcatttatt ctgcctccca
      721 gagcctgata aaaacggtta gcgcttcgtt aatacagatg taggtgttcc acagggtagc
      781 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgcttg tttcggcgtg
      841 ggtatggtgg caggccccgt ggccggggga ctgttgggcg ctgccggcac ctgtcctacg
      901 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
      961 cggaaggagc taccggacag cggtgcggac tgttgtaact cagaataaga aatgaggccg
     1021 ctcatggcgt tgactctcag tcatagtatc gtggtatcac cggttggttc cactctctgt
     1081 tgcgggcaac ttcagcagca cgtaggggac ttccgcgttt ccagacttta cgaaacacgg
     1141 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     1201 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     1261 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg
     1321 cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa
     1381 gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg
     1441 gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg
     1501 caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca
     1561 acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc ggtccagtga
     1621 tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat
     1681 ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa
     1741 gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag acgtagccca
     1801 gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     1861 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     1921 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     1981 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     2041 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgac
     2101 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     2161 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     2221 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     2281 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     2341 tgatgccggc cacgatgcgt ccggcgtaga ggatcagact aagatctcga tcccgcgaaa
     2401 ttaatacgac tcactatagg ggaattgtga gcggataaca attccccact agtaataatt
     2461 ttgtttaact ttaagaagga gatataccat gggcagcagc catcatcatc atcatcacag
     2521 cagcggtacc ggcagcggcg aaaaccttta cttccagggc catatgctcg aggacccaaa
     2581 gcagttggct gtttacgaag agtttgcacg caatgttcct ggcttcttac ctacaaatga
     2641 cttaagtcag cccacgggat ttttagccca gcccatgaag caagcttggg caacagatga
     2701 tgtagctcag atttatgata agtgtattac agaactggag caacatctac atgccatccc
     2761 accaactttg gccatgaacc ctcaagctca ggctcttcga agtctcttgg aggttgtagt
     2821 tttatctcga aactctcggg atgccatagc tgctcttgga ttgctccaaa aggctgtaga
     2881 gggcttacta gatgccacaa gtggtgctga tgctgacctt ctgctgcgct acagggaatg
     2941 ccacctcttg gtcctaaaag ctctgcagga tggccgggca tatgggtctc catggtgcaa
     3001 caaacagatc acaaggtgcc taattgaatg tcgagatgaa tataaatata atgtggaggc
     3061 tgtggagctg ctaattcgca atcatttggt taatatgcag cagtatgatc ttcacctagc
     3121 gcagtcaatg gagaatggct taaactacat ggctgtggca tttgctatgc agttagtaaa
     3181 aatcctgctg gtggatgaaa ggagtgttgc tcatgttact gaggcagatc tgttccacac
     3241 cattgaaacc ctcatgagga ttaatgctca ttccagaggc aatgctccag aaggattgcc
     3301 ccagctgatg gaagtagtgc gatccaacta tgaagcaatg attgatcgtg ctcatggagg
     3361 cccaaacttt atgatgcatt ctgggatctc tcaagcctca gagtatgatg accctccagg
     3421 cctgagggag aaggcagagt atcttctgag ggaatgggtg aatctctacc attcagcagc
     3481 agctggccgc gacagtacca aagctttctc tgcatttgtt ggacagatgc accagcaagg
     3541 aatactgaag accgatgatc tcataacaag gttctttcgt ctgtgtactg aaatgtgtgt
     3601 tgaaatcagt taccgtgctc aggctgagca gcagcacaat cctgctgcca atcccaccat
     3661 gatccgagcc aagtgctatc acaacctgga tgcctttgtt cgactcattg cactgctcgt
     3721 gaaacactca ggggaggcca ccaacactgt cacaaagatt aatctgctga acaaggtcct
     3781 tggtatagta gtgggagttc tccttcagga tcatgatgtt cgtcagagtg aatttcagca
     3841 acttccctac catcgaattt ttatcatgct tctcttggaa ctcaatgcac ctgagcatgt
     3901 gttggaaacc attaatttcc agacacttac agctttctgc aatacattcc acatcttgag
     3961 gcctaccaaa gctcctggct ttgtatatgc ctggcttgaa ctgatttccc atcggatatt
     4021 tattgcaaga atgctggcac atacgccaca gcagaagggg tggcctatgt atgcacagct
     4081 actgattgat ttattcaaat atttagcgcc tttccttaga aatgtggaac tcaccaaacc
     4141 tatgcaaatc ctctacaagg gcactttaag agtgctgctg gttcttttgc atgatttccc
     4201 agagttcctt tgtgattacc attatgggtt ctgtgatgtg atcccaccta attgtatcca
     4261 gttaagaaat ttgatcctga gtgcctttcc aagaaacatg aggctccccg acccattcac
     4321 tcctaatcta aaggtggaca tgttgagtga aattaacatt gctccccgga ttctcaccaa
     4381 tttcactgga gtaatgccac ctcagttcaa aaaggatttg gattcctatc ttaaaactcg
     4441 atcaccagtc actttcctgt ctgatctgcg cagcaaccta caggtatcca atgaacctgg
     4501 gaatcgctac aacctccagc tcatcaatgc actggtgctc tatgtcggga ctcaggccat
     4561 tgcgcacatc cacaacaagg gcagcacacc ttcaatgagc accatcactc actcagcaca
     4621 catggatatc ttccagaatt tggctgtgga cttggacact gagggtcgct atctcttttt
     4681 gaatgcaatt gcaaatcagc tccggtaccc aaatagccac actcactact tcagttgcac
     4741 catgctgtac ctttttgcag aggccaatac ggaagccatc caagaacaga tcacaagagt
     4801 tctcttggaa cggttgattg taaataggcc acatccttgg ggtcttctta ttaccttcat
     4861 tgagctgatt aaaaacccag cgtttaagtt ctggaaccat gaatttgtac actgtgcccc
     4921 agaaatcgaa aagttattcc agtcggtcgc acagtgctgc atgggacaga agcaggccca
     4981 gcaagtaatg gaataaggat cctaatagtc tagagctagc cctaggagat ccggctgcta
     5041 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     5101 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     5161 gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta cagcatccag
     5221 ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg gtgcctgact
     5281 gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga taagctgtca
     5341 aacatgagaa ttacaactta tatcgtatgg ggctgacttc aggtgctaca tttgaagaga
     5401 taaattgcac tgaaatcaag aaatatttta tctgattaat aagatgatct tcttgagatc
     5461 gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt
     5521 tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt ggaggagcgc
     5581 agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc aagactaact
     5641 cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg
     5701 gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc
     5761 atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg tggaatgaga
     5821 caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa caggagagcg
     5881 cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5941 ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa
     6001 cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc ttccaggaaa
     6061 tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga gcgtagcgag
     6121 tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg caccggtgca
     6181 gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg ccaacatagt
     6241 aagccagtat acactcccct agcgctgatg tccggcggtg cttttgccgt tacgcaccac
     6301 cccgtcagta gctgaacagg agggacagct gatagaaaca gaagccactg gagcacctca
     6361 aaaacaccat catacactaa atcagtaagt tggcagcatc acccgacgca ctttgcgccg
     6421 aataaatacc tgtgacggaa gatcacttcg cagaataaat aaatcctggt gtccctgttg
     6481 ataccgggaa gccctgggcc aacttttggc gaaaatgaga cgttgatcgg cacgtaagag
     6541 gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga
     6601 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     6661 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     6721 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     6781 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     6841 aattc
//
