LOCUS       sequence-290176-        5978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of CtNot1optim_1676-2193.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5978)
  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 5978)
  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-CtNot1optim_1676-2193_U
FEATURES             Location/Qualifiers
     source          1..5978
                     /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(4210..4228)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4224..4271
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4446..4451,4452..4468,4469..4474))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      4586..5131
                     /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     5032..5051
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5248..5265
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5657..5759
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(5760..5978,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5826..5845)
                     /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 aggcagcaca
     2581 tctgctggaa caaatggaat atgtttttga cgaatggatc cacctgtgca ataatccgca
     2641 tgcaaccgaa cgtgcagcaa tgatttttgt tcatcagctg catagcgttc agctggttac
     2701 caatcgtgat gaatttctgc tgttcctgcg tcatgcactg gataaaagcg ttgaacgttt
     2761 tgaacagggt attcatagcg gtgcaagcat tgcagaaagt tttcaggcag ttgaagcact
     2821 ggttaaactg atcatcatct ttgtgaaaag ccaccaggat agcgaagata aagatccgag
     2881 cgcagcagtt gcatttatgg atagcattct ggcactgggt gttctggttg caaatagcca
     2941 tcatgttaaa cgtggcgaaa actttaatca gcgtgtgttt tatcgttttt ttgcgctgct
     3001 gctgcatgaa gttggtctgc tggcaggtca ttttagcaaa agccattatg agcagatcat
     3061 cctgaatttt gcagcacgtc tgtttgatat gcgtccgaac ctgctgcctg gttttgcatg
     3121 tgcatgggca ggtctggtta gccatcgtgc atttctgccg gttattctgg gtctgccgga
     3181 tgaaaaaggt tgggcaccgt ttaccaaact gctggaacag tttctgggtt gtgttggtga
     3241 actggtgaaa acctttaccg ttagcagtct gggtaaagaa atgtatcatg cagccctgaa
     3301 aattctgatt gttctgcagc acgattttcc gatctatctg gataaatttc gtgttcagct
     3361 gtgtcagagc ctgccgctgc atgccaccca gctggtgaat ctgattctgg cagcaattcc
     3421 gcctaattgt aatagcctgg cagatccgtt tcaagcaggt ctgaaagttg ataaaatccc
     3481 ggatatgaaa gaacgtccgc ctaccgcatt tgatagcgca ggtctgctgc gtgaagcagg
     3541 actgctggat attctggaac gtatgctgca gaatggtccg agcgaagatg gtgttgcaca
     3601 gattaatcat gccattaaca aaagtgatgg tgagagcacc agctttggtt atgttccgct
     3661 gggtgttaat cgtcgtctga ttgatgcagt tgttgcacgt tttgcagaat ttgcaattaa
     3721 tcgtgccagc agccgtagcg atagcgcaat ttttgttgcc ggtgcaaatg atatcaaaac
     3781 cctgcagatg ctggtgaccg aagttagtcc ggaagcacgt tattatctgg ttagtagcat
     3841 ggttaatgaa ctgcgttatc cgaatgccta taccaactat tttagccagg ccctgctgga
     3901 catttttggt catgatatga gcgatccgga agaaaatctg gttcgtgagc agattgttcg
     3961 tgtcctgctg gaacgcgtgc tgggttattg gcctcagccg tggggtctga ttattaccat
     4021 cctggaactg ctgaaaaacg acaaatacct gttttttgaa ctgccgttca ttaaagcaac
     4081 accggaagtt gccgaacgtt ttaccgcact ggcacgtagc gcagcataag gatcctaata
     4141 gtctagagct agccctagga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     4201 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     4261 ggggtttttt gctgaaagga ggaactatat ccggatatcc cgcaagaggc ccggcagtac
     4321 cggcataacc aagcctatgc ctacagcatc cagggtgacg gtgccgagga tgacgatgag
     4381 cgcattgtta gatttcatac acggtgcctg actgcgttag caatttaact gtgataaact
     4441 accgcattaa agcttatcga tgataagctg tcaaacatga gaattacaac ttatatcgta
     4501 tggggctgac ttcaggtgct acatttgaag agataaattg cactgaaatc aagaaatatt
     4561 ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct
     4621 ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact
     4681 ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc ctttcagttt
     4741 agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc agtggctgct
     4801 gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt accggataag
     4861 gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga gcgaactgcc
     4921 tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca gcggaatgac
     4981 accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag ggggaaacgc
     5041 ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg
     5101 atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc tctcacttcc
     5161 ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta agccatttcc
     5221 gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc
     5281 ctgtatcaca tattctgctg acgcaccggt gcagcctttt ttctcctgcc acatgaagca
     5341 cttcactgac accctcatca gtgccaacat agtaagccag tatacactcc cctagcgctg
     5401 atgtccggcg gtgcttttgc cgttacgcac caccccgtca gtagctgaac aggagggaca
     5461 gctgatagaa acagaagcca ctggagcacc tcaaaaacac catcatacac taaatcagta
     5521 agttggcagc atcacccgac gcactttgcg ccgaataaat acctgtgacg gaagatcact
     5581 tcgcagaata aataaatcct ggtgtccctg ttgataccgg gaagccctgg gccaactttt
     5641 ggcgaaaatg agacgttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa
     5701 gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa
     5761 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
     5821 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
     5881 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
     5941 acattcttgc ccgcctgatg aatgctcatc cggaattc
//
