LOCUS       sequence-291121-        6980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsNot1_1093-1588.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6980)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6980)
  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"
COMMENT     Sequence Label: pnYC-NpM-HsNot1_1093-1588_AF
FEATURES             Location/Qualifiers
     source          1..6980
                     /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             2489..3589
                     /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             3605..3628
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(5212..5230)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5226..5273
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5448..5453,5454..5470,5471..5476))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      5588..6133
                     /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     6034..6053
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6250..6267
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        6659..6761
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(6762..6980,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6828..6847)
                     /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 ggaaactgaa gaaggtaaac tggtaatctg
     2521 gattaacggc gataaaggct ataacggtct cgctgaagtc ggtaagaaat tcgagaaaga
     2581 taccggaatt aaagtcaccg ttgagcatcc ggataaactg gaagagaaat tcccacaggt
     2641 tgcggcaact ggcgatggcc ctgacattat cttctgggca cacgaccgct ttggtggcta
     2701 cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa gcgttccagg acaagctgta
     2761 tccgtttacc tgggatgccg tacgttacaa cggcaagctg attgcttacc cgatcgctgt
     2821 tgaagcgtta tcgctgattt ataacaaaga cctgctgccg aacccgccaa aaacctggga
     2881 agagatcccg gcgctggata aagaactgaa agcgaaaggt aagagcgccc tgatgttcaa
     2941 cctgcaagaa ccgtacttca cctggccgct gattgctgct gacgggggtt atgcgttcaa
     3001 gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg gataacgctg gcgcgaaagc
     3061 gggtctgacc ttcctggttg acctgattaa aaacaaacac atgaatgcag acaccgatta
     3121 ctccatcgca gaagctgcct ttaataaagg cgaaacagcg atgaccatca acggcccgtg
     3181 ggcatggtcc aacatcgaca ccagcaaagt gaattatggt gtaacggtac tgccgacctt
     3241 caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc gcaggtatta acgccgccag
     3301 tccgaacaaa gagctggcaa aagagttcct cgaaaactat ctgctgactg atgaaggtct
     3361 ggaagcggtt aataaagaca aaccgctggg tgccgtagcg ctgaagtctt acgaggaaga
     3421 gttggcgaaa gatccacgta ttgccgccac gatggaaaac gcccagaaag gtgaaatcat
     3481 gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg cgtactgcgg tgatcaacgc
     3541 cgccagcggt cgtcagactg tcgatgaagc ccttaaagac gcgcagactg gtaccggcag
     3601 cggcctggaa gttctgttcc aggggcccca tatgctcgag gaaaatatcc aggagaaaat
     3661 tgcttttatt ttcaataatc tctcacagtc aaatatgaca caaaaggttg aagagctaaa
     3721 ggaaacggtg aaagaagaat ttatgccttg ggtttcacag tatctggtta tgaagagagt
     3781 cagtattgag ccaaactttc atagcctgta ttcaaacttc cttgacacgc tgaagaatcc
     3841 tgaatttaac aagatggttc tgaatgagac ctacagaaac attaaagtgc tcctgacctc
     3901 tgataaagct gcagccaatt tctcagatcg ttctttgctg aagaacttgg gacattggct
     3961 aggaatgatc acattagcta aaaacaaacc catcttacac actgacttgg atgtgaaatc
     4021 attgctgcta gaggcttatg ttaaaggaca acaagaattg ctctatgtag tgccctttgt
     4081 tgccaaagtc ttagaatcta gcattcgtag tgtggttttt aggccaccaa acccttggac
     4141 aatggcaatt atgaatgtat tagctgagct acatcaggag catgacttaa agttaaactt
     4201 gaagtttgaa atcgaggttc tctgcaagaa ccttgcatta gacatcaatg agctaaaacc
     4261 tggaaacctc ctaaaggata aagatcgcct gaagaattta gatgagcaac tctctgctcc
     4321 aaagaaagat gtcaagcagc cagaagaact ccctcccatc acaaccacaa caacttctac
     4381 tacaccagct accaacacca cttgtacagc cacggttcca ccacagccac agtacagcta
     4441 ccacgacatc aatgtctatt cccttgcggg cttggcacca cacattactc taaatccaac
     4501 aattcccttg tttcaggccc atccacagtt gaagcagtgt gtgcgtcagg caattgaacg
     4561 ggctgtccag gagctggtcc atcctgtggt ggatcgatca attaagattg ccatgactac
     4621 ttgtgagcaa atagtcagga aggattttgc cctggattcg gaggaatctc gaatgcgaat
     4681 agcagctcat cacatgatgc gtaacttgac agctggaatg gctatgatta catgcaggga
     4741 acctttgctc atgagcatat ctaccaactt aaaaaacagt tttgcctcag cccttcgtac
     4801 tgcttcccca caacaaagag aaatgatgga tcaggcagct gctcaattag ctcaggacaa
     4861 ttgtgagttg gcttgctgtt ttattcagaa gactgcagta gaaaaagcag gccctgagat
     4921 ggacaagaga ttagcaactg aatttgagct gagaaaacat gctaggcaag aaggacgcag
     4981 atactgtgat cctgttgttt taacatatca agctgaacgg atgccagagc aaatcaggct
     5041 gaaagttggt ggtgtggacc caaagcagtt ggctgtttac gaagagtttg cacgcaatgt
     5101 tcctggcttc ttacctacaa atgacttata aggatcctaa tagtctagag ctagccctag
     5161 gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     5221 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     5281 gaggaactat atccggatat cccgcaagag gcccggcagt accggcataa ccaagcctat
     5341 gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat
     5401 acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc
     5461 gatgataagc tgtcaaacat gagaattaca acttatatcg tatggggctg acttcaggtg
     5521 ctacatttga agagataaat tgcactgaaa tcaagaaata ttttatctga ttaataagat
     5581 gatcttcttg agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg
     5641 ccttgcaggg cggtttttcg aaggttctct gagctaccaa ctctttgaac cgaggtaact
     5701 ggcttggagg agcgcagtca ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg
     5761 acttcaagac taactcctct aaatcaatta ccagtggctg ctgccagtgg tgcttttgca
     5821 tgtctttccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcggactga
     5881 acggggggtt cgtgcataca gtccagcttg gagcgaactg cctacccgga actgagtgtc
     5941 aggcgtggaa tgagacaaac gcggccataa cagcggaatg acaccggtaa accgaaaggc
     6001 aggaacagga gagcgcacga gggagccgcc agggggaaac gcctggtatc tttatagtcc
     6061 tgtcgggttt cgccaccact gatttgagcg tcagatttcg tgatgcttgt caggggggcg
     6121 gagcctatgg aaaaacggct ttgccgcggc cctctcactt ccctgttaag tatcttcctg
     6181 gcatcttcca ggaaatctcc gccccgttcg taagccattt ccgctcgccg cagtcgaacg
     6241 accgagcgta gcgagtcagt gagcgaggaa gcggaatata tcctgtatca catattctgc
     6301 tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag cacttcactg acaccctcat
     6361 cagtgccaac atagtaagcc agtatacact cccctagcgc tgatgtccgg cggtgctttt
     6421 gccgttacgc accaccccgt cagtagctga acaggaggga cagctgatag aaacagaagc
     6481 cactggagca cctcaaaaac accatcatac actaaatcag taagttggca gcatcacccg
     6541 acgcactttg cgccgaataa atacctgtga cggaagatca cttcgcagaa taaataaatc
     6601 ctggtgtccc tgttgatacc gggaagccct gggccaactt ttggcgaaaa tgagacgttg
     6661 atcggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat
     6721 tttttgagtt atcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
     6781 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     6841 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
     6901 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     6961 tgaatgctca tccggaattc
//
