LOCUS       sequence-291187-        7316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsNot1_1-1000.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7316)
  AUTHORS   Sgromo A, Raisch T, Backhaus C, Keskeny C, Alva V, Weichenrieder O, 
            Izaurralde E
  TITLE     Drosophila Bag-of-marbles directly interacts with the CAF40 subunit 
            of the CCR4-NOT complex to elicit repression of mRNA targets.
  JOURNAL   RNA. 2018 Mar;24(3):381-395. doi: 10.1261/rna.064584.117. Epub 2017 
            Dec 18.
  PUBMED    29255063
REFERENCE   2  (bases 1 to 7316)
  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: 
            "10.1261/rna.064584.117"; journalName: "RNA"; date: "2018-03"; 
            volume: "24"; issue: "3"; pages: "381-395"
COMMENT     Sequence Label: pnYC-HsNot1_1-1000_AF
FEATURES             Location/Qualifiers
     source          1..7316
                     /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)"
     primer_bind     complement(5548..5566)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5562..5609
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5784..5789,5790..5806,5807..5812))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      5924..6469
                     /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     6370..6389
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6586..6603
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        6995..7097
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(7098..7316,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(7164..7183)
                     /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 gaacctggac tcgctctcgc tggccttgtc
     2521 tcaaatcagc tacctggtgg acaatttaac caagaaaaat taccgagcca gccagcagga
     2581 aatacagcat attgtgaatc ggcacggtcc tgaggcagac aggcatttat tacgctgcct
     2641 attttcgcat gtggatttca gtggcgatgg taaaagcagt ggcaaagatt tccatcagac
     2701 tcagtttctg attcaggagt gtgcgttgct gattacaaag ccaaatttta tctcgacgct
     2761 gtcctatgcc attgataatc cattgcacta tcagaagagt ttaaagcctg caccccactt
     2821 atttgcccag ctgagtaaag tgctcaaatt aagcaaagta caagaggtaa tttttggcct
     2881 tgccctgttg aattcttcca gctcagatct tagaggtttc gctgcccagt ttatcaaaca
     2941 gaagcttcca gatcttctgc gttcttacat tgacgcagac gtcagtggaa atcaagaagg
     3001 tggcttccaa gatatagcaa tagaggtcct acacctcctc ctctcccatc tcctctttgg
     3061 gcagaaggga gcctttggag ttggacaaga acagatagac gcttttctta agacgctgcg
     3121 cagagatttt ccccaagaac gctgtcccgt ggtgctcgca ccacttttat accctgaaaa
     3181 acgggacatt ctaatggaca ggatcctgcc tgattccgga ggggtagcta aaaccatgat
     3241 ggagagctct ttggctgatt tcatgcaaga agtaggctat ggcttttgtg caagtattga
     3301 agaatgtcgc aatataatcg tgcagtttgg tgttcgggag gtcacagctg cccaggttgc
     3361 aagggttttg ggaatgatgg ctcgaactca ttcaggatta acagatggca ttccattaca
     3421 gagtatttct gctccgggca gtgggatctg gagtgatggg aaagataaaa gtgatggagc
     3481 acaggcacac acatggaatg tagaagtctt gattgacgtt cttaaagaac tgaatccaag
     3541 tttgaatttc aaggaagtaa cttatgaact ggaccatcct ggatttcaaa ttcgtgacag
     3601 taaaggactt cataatgtgg tttatggcat tcagaggggt ttgggtatgg aagtgttccc
     3661 agtagacctc atatatagac cttggaaaca tgctgaaggc cagctctcct tcattcaaca
     3721 ttcccttata aatccagaga tcttctgttt tgctgactat ccctgtcata ctgttgccac
     3781 tgatattctg aaagcaccac cagaggatga caatcgagaa attgccacat ggaagagctt
     3841 ggatttgatt gaatctctgc tgaggcttgc agaggttggg cagtatgagc aagtcaaaca
     3901 gctcttcagc ttccctatca aacactgtcc agacatgctg gtattggcct tactacaaat
     3961 taacacctct tggcatacct tgcgccatga acttatctcc actctgatgc caattttcct
     4021 tggaaaccat cctaactcag ctattatttt gcactatgca tggcatgggc agggacagtc
     4081 tccctcaatt cgccaactta tcatgcatgc aatggcagaa tggtacatga gaggggagca
     4141 gtatgatcag gccaaattgt ctcgaatact tgatgtggcc caggacttga aggccttgtc
     4201 aatgctgcta aatggtactc catttgcctt tgttattgac cttgctgcac ttgcttcacg
     4261 tcgtgaatac ctcaaacttg ataagtggct cacagataaa attcgagagc atggggagcc
     4321 ttttatccag gcgtgtatga cttttttaaa gagacggtgt ccttctattt tgggcggact
     4381 tgccccagaa aaagaccagc ccaaaagtgc tcaacttcct ccagaaactt tggcgacaat
     4441 gttggcctgt ctgcaagctt gtgcagggag tgtttctcag gagctatcag aaactatcct
     4501 caccatggta gccaattgca gtaatgttat gaataaggcc agacaaccac cacctggagt
     4561 tatgccaaaa ggacgtcctc ctagtgctag cagcttagat gccatttctc ctgttcagat
     4621 tgaccctctt gctggaatga catctcttag tataggtggt tcagctgccc ctcacaccca
     4681 gagtatgcag ggttttcctc caaatttggg ttctgcattc agtacccctc agtcaccagc
     4741 aaaagcattt ccaccccttt caacccccaa tcagaccact gcattcagtg gtattggagg
     4801 actttcatca cagcttccag taggtggtct tggcacaggc agcctgactg gtataggaac
     4861 tggtgctctt ggactccctg cagtgaataa cgaccctttt gtacagagga aactgggcac
     4921 ctctggactg aatcagccta cattccagca gagtaagatg aaaccttcgg acttgtctca
     4981 ggtgtggcca gaggcaaacc agcactttag taaagagata gatgatgaag caaacagcta
     5041 tttccagcga atatataatc atccaccaca tccaaccatg tctgttgatg aggtattaga
     5101 aatgctgcag agatttaaag actctactat aaagagggaa cgagaagtat ttaactgtat
     5161 gctaaggaac ttgtttgaag aatatcgttt ttttccccag tatcctgata aagagttaca
     5221 tataacagcc tgcctatttg gtggtataat tgagaaagga ctggtcactt acatggcact
     5281 aggtctggct ctacgatatg ttcttgaagc cttacgcaag ccttttggat ccaaaatgta
     5341 ttatttcggg attgctgcac tagatagatt taaaaacaga ttgaaggact atccccagta
     5401 ttgtcagcat ttggcttcta tcagtcactt tatgcaattt ccacatcatt tacaggagta
     5461 tattgagtat ggacagcagt ctagagatta acctaggaga tccggctgct aacaaagccc
     5521 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     5581 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatatcccg
     5641 caagaggccc ggcagtaccg gcataaccaa gcctatgcct acagcatcca gggtgacggt
     5701 gccgaggatg acgatgagcg cattgttaga tttcatacac ggtgcctgac tgcgttagca
     5761 atttaactgt gataaactac cgcattaaag cttatcgatg ataagctgtc aaacatgaga
     5821 attacaactt atatcgtatg gggctgactt caggtgctac atttgaagag ataaattgca
     5881 ctgaaatcaa gaaatatttt atctgattaa taagatgatc ttcttgagat cgttttggtc
     5941 tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt ttttcgaagg
     6001 ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg cagtcaccaa
     6061 aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac tcctctaaat
     6121 caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt ggactcaaga
     6181 cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg catacagtcc
     6241 agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag acaaacgcgg
     6301 ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc gcacgaggga
     6361 gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc accactgatt
     6421 tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa acggctttgc
     6481 cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa atctccgccc
     6541 cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga gtcagtgagc
     6601 gaggaagcgg aatatatcct gtatcacata ttctgctgac gcaccggtgc agcctttttt
     6661 ctcctgccac atgaagcact tcactgacac cctcatcagt gccaacatag taagccagta
     6721 tacactcccc tagcgctgat gtccggcggt gcttttgccg ttacgcacca ccccgtcagt
     6781 agctgaacag gagggacagc tgatagaaac agaagccact ggagcacctc aaaaacacca
     6841 tcatacacta aatcagtaag ttggcagcat cacccgacgc actttgcgcc gaataaatac
     6901 ctgtgacgga agatcacttc gcagaataaa taaatcctgg tgtccctgtt gataccggga
     6961 agccctgggc caacttttgg cgaaaatgag acgttgatcg gcacgtaaga ggttccaact
     7021 ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag
     7081 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
     7141 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     7201 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     7261 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattc
//
