LOCUS       sequence-358005-        5765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsNot1iso1_1351-1588.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5765)
  AUTHORS   Sgromo A, Raisch T, Bawankar P, Bhandari D, Chen Y, Kuzuoglu-Ozturk 
            D, Weichenrieder O, Izaurralde E
  TITLE     A CAF40-binding motif facilitates recruitment of the CCR4-NOT 
            complex to mRNAs targeted by Drosophila Roquin.
  JOURNAL   Nat Commun. 2017 Feb 6;8:14307. doi: 10.1038/ncomms14307.
  PUBMED    28165457
REFERENCE   2  (bases 1 to 5765)
  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: "Nat 
            Commun."; date: "2017-02-6"; volume: "8:14307. doi"; pages: " 
            10.1038/ncomms14307"
COMMENT     Sequence Label: pnYC-NpM-HsNot1iso1_1351-1588_AE
FEATURES             Location/Qualifiers
     source          1..5765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1853..1872
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1961..1980
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1961..1979
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1980..2004
                     /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             2019..2041
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2048..3148
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="METEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     CDS             3164..3187
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3997..4015)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4011..4058
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4233..4238,4239..4255,4256..4261))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      4373..4918
                     /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     4819..4838
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5035..5052
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5444..5546
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(5547..5765,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5613..5632)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 cgtatggcaa 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 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
      421 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt
      481 tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgaa agcaaattcg
      541 acccggtcgt cggttcaggg cagggtcgtt aaatagccgc ttatgtctat tgctggttta
      601 ccggtttatt gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc tgaggccagt
      661 ttgctcaggc tctccccgtg gaggtaataa ttgacgatat gatcatttat tctgcctccc
      721 agagcctgat aaaaacggtt agcgcttcgt taatacagat gtaggtgttc cacagggtag
      781 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctt gtttcggcgt
      841 gggtatggtg gcaggccccg tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg
      901 ctgctggaga tggcggacgc gatggatatg ttctgccaag ggttggtttg cgcattcaca
      961 gttctccgca agaattgatt ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc
     1021 cgccggcttc cattcaggtc gaggtggccc ggctccatgc accgcgacgc aacgcgggga
     1081 ggcagacaag gtatagggcg gcgcctacaa tccatgccaa cccgttccat gtgctcgccg
     1141 aggcggcata aatcgccgtg acgatcagcg gtccaatgat cgaagttagg ctggtaagag
     1201 ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg
     1261 cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca
     1321 tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg
     1381 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
     1441 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
     1501 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
     1561 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
     1621 agctgactgg gttgaaggct ctcaagggca tcggtcgacg ctctccctta tgcgactcct
     1681 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     1741 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     1801 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     1861 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     1921 cggcgtagag gatcagacta agatctcgat cccgcgaaat taatacgact cactataggg
     1981 gaattgtgag cggataacaa ttccccacta gtaataattt tgtttaactt taagaaggag
     2041 atataccatg gaaactgaag aaggtaaact ggtaatctgg attaacggcg ataaaggcta
     2101 taacggtctc gctgaagtcg gtaagaaatt cgagaaagat accggaatta aagtcaccgt
     2161 tgagcatccg gataaactgg aagagaaatt cccacaggtt gcggcaactg gcgatggccc
     2221 tgacattatc ttctgggcac acgaccgctt tggtggctac gctcaatctg gcctgttggc
     2281 tgaaatcacc ccggacaaag cgttccagga caagctgtat ccgtttacct gggatgccgt
     2341 acgttacaac ggcaagctga ttgcttaccc gatcgctgtt gaagcgttat cgctgattta
     2401 taacaaagac ctgctgccga acccgccaaa aacctgggaa gagatcccgg cgctggataa
     2461 agaactgaaa gcgaaaggta agagcgccct gatgttcaac ctgcaagaac cgtacttcac
     2521 ctggccgctg attgctgctg acgggggtta tgcgttcaag tatgaaaacg gcaagtacga
     2581 cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg ggtctgacct tcctggttga
     2641 cctgattaaa aacaaacaca tgaatgcaga caccgattac tccatcgcag aagctgcctt
     2701 taataaaggc gaaacagcga tgaccatcaa cggcccgtgg gcatggtcca acatcgacac
     2761 cagcaaagtg aattatggtg taacggtact gccgaccttc aagggtcaac catccaaacc
     2821 gttcgttggc gtgctgagcg caggtattaa cgccgccagt ccgaacaaag agctggcgaa
     2881 agagttcctc gaaaactatc tgctgactga tgaaggtctg gaagcggtta ataaagacaa
     2941 accgctgggt gccgtagcgc tgaagtctta cgaggaagag ttggcgaaag atccacgtat
     3001 tgccgccacg atggaaaacg cccagaaagg tgaaatcatg ccgaacatcc cgcagatgtc
     3061 cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc gccagcggtc gtcagactgt
     3121 cgatgaagcc cttaaagacg cgcagactgg taccggcagc ggcctggaag ttctgttcca
     3181 ggggccccat atgctcgagg ttccaccaca gccacagtac agctaccacg acatcaatgt
     3241 ctattccctt gcgggcttgg caccacacat tactctaaat ccaacaattc ccttgtttca
     3301 ggcccatcca cagttgaagc agtgtgtgcg tcaggcaatt gaacgggctg tccaggagct
     3361 ggtccatcct gtggtggatc gatcaattaa gattgccatg actacttgtg agcaaatagt
     3421 caggaaggat tttgccctgg attcggagga atctcgaatg cgaatagcag ctcatcacat
     3481 gatgcgtaac ttgacagctg gaatggctat gattacatgc agggaacctt tgctcatgag
     3541 catatctacc aacttaaaaa acagttttgc ctcagccctt cgtactgctt ccccacaaca
     3601 aagagaaatg atggatcagg cagctgctca attagctcag gacaattgtg agttggcttg
     3661 ctgttttatt cagaagactg cagtagaaaa agcaggccct gagatggaca agagattagc
     3721 aactgaattt gagctgagaa aacatgctag gcaagaagga cgcagatact gtgatcctgt
     3781 tgttttaaca tatcaagctg aacggatgcc agagcaaatc aggctgaaag ttggtggtgt
     3841 ggacccaaag cagttggctg tttacgaaga gtttgcacgc aatgttcctg gcttcttacc
     3901 tacaaatgac ttataaggat cctaatagtc tagagctagc cctaggagat ccggctgcta
     3961 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     4021 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     4081 gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta cagcatccag
     4141 ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg gtgcctgact
     4201 gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga taagctgtca
     4261 aacatgagaa ttacaactta tatcgtatgg ggctgacttc aggtgctaca tttgaagaga
     4321 taaattgcac tgaaatcaag aaatatttta tctgattaat aagatgatct tcttgagatc
     4381 gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt
     4441 tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt ggaggagcgc
     4501 agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc aagactaact
     4561 cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg
     4621 gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc
     4681 atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg tggaatgaga
     4741 caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa caggagagcg
     4801 cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4861 ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa
     4921 cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc ttccaggaaa
     4981 tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga gcgtagcgag
     5041 tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg caccggtgca
     5101 gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg ccaacatagt
     5161 aagccagtat acactcccct agcgctgatg tccggcggtg cttttgccgt tacgcaccac
     5221 cccgtcagta gctgaacagg agggacagct gatagaaaca gaagccactg gagcacctca
     5281 aaaacaccat catacactaa atcagtaagt tggcagcatc acccgacgca ctttgcgccg
     5341 aataaatacc tgtgacggaa gatcacttcg cagaataaat aaatcctggt gtccctgttg
     5401 ataccgggaa gccctgggcc aacttttggc gaaaatgaga cgttgatcgg cacgtaagag
     5461 gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga
     5521 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     5581 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     5641 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     5701 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     5761 aattc
//
