LOCUS       sequence-290419-        8231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of CtNot1_925-2193.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8231)
  AUTHORS   Raisch T, Sandmeir F, Weichenrieder O, Valkov E, Izaurralde E
  TITLE     Structural and biochemical analysis of a NOT1 MIF4G-like domain of 
            the CCR4-NOT complex.
  JOURNAL   J Struct Biol. 2018 Dec;204(3):388-395. doi: 
            10.1016/j.jsb.2018.10.009. Epub 2018 Oct 24.
  PUBMED    30367941
REFERENCE   2  (bases 1 to 8231)
  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: 
            "10.1016/j.jsb.2018.10.009"; journalName: "J Struct Biol"; date: 
            "2018-12"; volume: "204"; issue: "3"; pages: "388-395"
COMMENT     Sequence Label: pnYC-NvH-CtNot1_925-2193_X
FEATURES             Location/Qualifiers
     source          1..8231
                     /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(6463..6481)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6477..6524
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6699..6704,6705..6721,6722..6727))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      6839..7384
                     /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     7285..7304
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7501..7518
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        7910..8012
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(8013..8231,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(8079..8098)
                     /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 agagcgaaga
     2581 agttcagggt aaagttcagt ttgtgctgaa taacctgacc gataaaacca tgaaaagcag
     2641 cagcaaagaa ctgaaaggta tgctggaaag caaacactat cagtggttta gcatgcatct
     2701 ggttgaagaa cgtgcaaaaa tgcagccgaa ttatcacaac gtgtatatcg aactggtgaa
     2761 acagttcgag gataaaaaac tgtgggcaga agttctgcgt cagacctttg ttagcgttca
     2821 gcgtatgctg aatagcgaaa gcaccatgca gaatagcacc gaacgtaatc atctgaaaaa
     2881 tctggcaggt tggctgggtc tgctgaccct ggcacgtgat aaaccgatta aacataaaaa
     2941 cattgccttt aaacaactgc tgatcgaagc ccatgatacc aaacgtctgg ttgttgttat
     3001 tccgtttgtt tgcaaagttc tgattgccgg tgcaaatagc aatgtgttta aaccgcctaa
     3061 tccgtggctg atggatatta ttcatctgct gattgagctg tatcataacg caagcctgaa
     3121 actgaacctg aaatttgaaa ttgaggtgct gtgtaaaggc ctgaatctgg atcacaatac
     3181 cattgaaccg agcggtgaaa ttctgaatcg tccgcctgcg gaagaggtta gcggtgatat
     3241 gctgcatccg gaaggtctgg aaaattttga aaatctgagc ctgaatggta tgcctggtgt
     3301 gaataccacc ctggcaagcc atccggcaat taccattccg gatctgggtc cgagcctgac
     3361 aattccgcag acagaagttg ttaataccgc aaaactgcat gaaattgttc gtcaggccct
     3421 gacccgtgca ctgcaggata tcattcagcc ggttgttgat cgtagcgtta ccattgcagc
     3481 aattagcacc catcagatga tccgtaaaga ttttgcaacc gaaccggatg aaaatcgtgt
     3541 tcgtaccgca gccattaata tggttaaaag caccgcaggt agcctggcac tggttaccag
     3601 taaagaaccg ctgcgtgcaa attttgcaaa ttatctgcgt aatctgagtg cagaactgcc
     3661 gcagggtctg ccggaaggca ttattatgct gtgtgttaat agcaacctgg atctggcaag
     3721 cagcgttatt gaaaaaagcg cagaagaacg cgcagttccg gaaattgaag aaatgctgga
     3781 accggaactg gaagcccgtc gtcgtcaccg tctgacccgt ccgaatgaac cgtatatcga
     3841 tcatggtctg agccgttggg ccatgaccat tccgcatccg tataaactgc agccgagcct
     3901 gcatggcctg aatgcagagc agatggcaat ttatgatgat tttgcccgtg gcacccgtat
     3961 gaataccgca ggcaccgttc cggcacatgt tagcaccgca agcgataccc gtagcattgc
     4021 aaatgaagca ctgagcgatc agtatagcgg tgttagtacc atgccgacac cggcagaaac
     4081 tccggcaatg ccgcatctgg gtgcacagat gcaacattat cctcaggcac atatgggtct
     4141 gagcaatggt cgtccgaccg gtctgaatca gattgatagc cgtgcagttg cagaacgcat
     4201 taacaaatat ctggaacagc tgaccgcagc cgcaaccagc gcaaccgaag aacattttaa
     4261 tgaactgcct cgtccgcatg cagttctgga cattattgat gccctgattc agctgattat
     4321 caaagcacag cagaccagcg aagaatttgc gatttatgca ctgcagcaga ttagccagct
     4381 gctgtttcgt cagccggaag gcaccctgct gctggaaagc ctggttcatg ttctggaaac
     4441 cattcgtaaa attgcaggtc cgcaggttag cgaacaggtt cgtcagctgt ttcaccagca
     4501 gcctggtcac ctgtttctga gcctgagtct gattgcagca ctgctgggca ccgatctgct
     4561 ggattggaaa aacattgata tggcaatggc aaaagcactg gaacagcgta aagaaggtag
     4621 cattgatttt ctggaacaac tgatggatct ggtgctgctg aatgatacac cgctggcact
     4681 gtttaccgat tttgttcgta gcctggaagc agcatgggca tggattgttg aagatcctga
     4741 tctgcctgca gcacagcgtt ttaaagcaaa agttcgtgca cagctgccgg aacctccggc
     4801 agcaaatctg agcaaagaag aacacgcagc acatctgctg gaacaaatgg aatatgtttt
     4861 tgacgaatgg attcacctgt gcaataatcc gcatgcaacc gaacgtgcag caatgatttt
     4921 tgttcatcag ctgcatagcg ttcagctggt taccaatcgt gatgaatttc tgctgttcct
     4981 gcgtcatgca ctggataaaa gcgttgaacg ttttgaacag ggtattcata gcggtgcaag
     5041 cattgcagaa agttttcagg cagttgaagc actggttaaa ctgatcatca tctttgtgaa
     5101 aagccaccag gatagcgaag ataaagatcc gagcgcagca gttgcattta tggatagcat
     5161 tctggcactg ggtgttctgg ttgcaaatag ccatcatgtt aaacgtggcg aaaactttaa
     5221 tcagcgtgtg ttttatcgtt tttttgcgct gctgctgcat gaagttggtc tgctggcagg
     5281 tcattttagc aaaagccatt atgagcagat catcctgaat tttgcagcac gtctgtttga
     5341 tatgcgtccg aacctgctgc ctggttttgc atgtgcatgg gcaggtctgg ttagccatcg
     5401 tgcatttctg ccggttattc tgggtctgcc ggatgaaaaa ggttgggcac cgtttaccaa
     5461 actgctggaa cagtttctgg gttgtgttgg tgaactggtg aaaaccttta ccgttagcag
     5521 tctgggtaaa gaaatgtatc atgcagccct gaaaattctg attgttctgc agcacgattt
     5581 tccgatctat ctggataaat ttcgtgttca gctgtgtcag agcctgccgc tgcatgccac
     5641 ccagctggtg aatctgattc tggcagcaat tccgcctaat tgtaatagcc tggcagatcc
     5701 gtttcaagca ggtctgaaag ttgataaaat cccggatatg aaagaacgtc cgcctaccgc
     5761 atttgatagc gcaggtctgc tgcgtgaagc aggactgctg gatattctgg aacgtatgct
     5821 gcagaatggt ccgagcgaag atggtgttgc acagattaat catgccatta acaaaagtga
     5881 tggtgagagc accagctttg gttatgttcc gctgggtgtt aatcgtcgtc tgattgatgc
     5941 agttgttgca cgttttgcag aatttgcaat taatcgtgcc agcagccgta gcgatagcgc
     6001 aatttttgtt gccggtgcaa atgatatcaa aaccctgcag atgctggtga ccgaagttag
     6061 tccggaagca cgttattatc tggttagtag catggttaat gaactgcgtt atccgaatgc
     6121 ctataccaac tattttagcc aggccctgct ggacattttt ggtcatgata tgagcgatcc
     6181 ggaagaaaat ctggttcgtg agcagattgt tcgtgtcctg ctggaacgcg tgctgggtta
     6241 ttggcctcag ccgtggggtc tgattattac catcctggaa ctgctgaaaa acgacaaata
     6301 cctgtttttt gaactgccgt tcattaaagc aacaccggaa gttgccgaac gttttaccgc
     6361 actggcacgt agcgcagcat aaggatccta atagtctaga gctagcccta ggagatccgg
     6421 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     6481 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     6541 tatccggata tcccgcaaga ggcccggcag taccggcata accaagccta tgcctacagc
     6601 atccagggtg acggtgccga ggatgacgat gagcgcattg ttagatttca tacacggtgc
     6661 ctgactgcgt tagcaattta actgtgataa actaccgcat taaagcttat cgatgataag
     6721 ctgtcaaaca tgagaattac aacttatatc gtatggggct gacttcaggt gctacatttg
     6781 aagagataaa ttgcactgaa atcaagaaat attttatctg attaataaga tgatcttctt
     6841 gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg
     6901 gcggtttttc gaaggttctc tgagctacca actctttgaa ccgaggtaac tggcttggag
     6961 gagcgcagtc accaaaactt gtcctttcag tttagcctta accggcgcat gacttcaaga
     7021 ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc atgtctttcc
     7081 gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg aacggggggt
     7141 tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt caggcgtgga
     7201 atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg caggaacagg
     7261 agagcgcacg agggagccgc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7321 tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc ggagcctatg
     7381 gaaaaacggc tttgccgcgg ccctctcact tccctgttaa gtatcttcct ggcatcttcc
     7441 aggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt
     7501 agcgagtcag tgagcgagga agcggaatat atcctgtatc acatattctg ctgacgcacc
     7561 ggtgcagcct tttttctcct gccacatgaa gcacttcact gacaccctca tcagtgccaa
     7621 catagtaagc cagtatacac tcccctagcg ctgatgtccg gcggtgcttt tgccgttacg
     7681 caccaccccg tcagtagctg aacaggaggg acagctgata gaaacagaag ccactggagc
     7741 acctcaaaaa caccatcata cactaaatca gtaagttggc agcatcaccc gacgcacttt
     7801 gcgccgaata aatacctgtg acggaagatc acttcgcaga ataaataaat cctggtgtcc
     7861 ctgttgatac cgggaagccc tgggccaact tttggcgaaa atgagacgtt gatcggcacg
     7921 taagaggttc caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt
     7981 tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
     8041 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
     8101 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
     8161 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
     8221 atccggaatt c
//
