LOCUS       sequence-291122-        7037 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsNot1_1093-1607.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7037)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7037)
  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_1093-1607_AF
FEATURES             Location/Qualifiers
     source          1..7037
                     /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(5269..5287)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5283..5330
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5505..5510,5511..5527,5528..5533))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      5645..6190
                     /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     6091..6110
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6307..6324
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        6716..6818
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(6819..7037,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6885..6904)
                     /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 atgacttaag tcagcccacg ggatttttag cccagcccat
     5161 gaagcaagct tgggcaacag atgattaagg atcctaatag tctagagcta gccctaggag
     5221 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     5281 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     5341 gaactatatc cggatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc
     5401 tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca
     5461 cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat
     5521 gataagctgt caaacatgag aattacaact tatatcgtat ggggctgact tcaggtgcta
     5581 catttgaaga gataaattgc actgaaatca agaaatattt tatctgatta ataagatgat
     5641 cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc tgaaaacgaa aaaaccgcct
     5701 tgcagggcgg tttttcgaag gttctctgag ctaccaactc tttgaaccga ggtaactggc
     5761 ttggaggagc gcagtcacca aaacttgtcc tttcagttta gccttaaccg gcgcatgact
     5821 tcaagactaa ctcctctaaa tcaattacca gtggctgctg ccagtggtgc ttttgcatgt
     5881 ctttccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc ggactgaacg
     5941 gggggttcgt gcatacagtc cagcttggag cgaactgcct acccggaact gagtgtcagg
     6001 cgtggaatga gacaaacgcg gccataacag cggaatgaca ccggtaaacc gaaaggcagg
     6061 aacaggagag cgcacgaggg agccgccagg gggaaacgcc tggtatcttt atagtcctgt
     6121 cgggtttcgc caccactgat ttgagcgtca gatttcgtga tgcttgtcag gggggcggag
     6181 cctatggaaa aacggctttg ccgcggccct ctcacttccc tgttaagtat cttcctggca
     6241 tcttccagga aatctccgcc ccgttcgtaa gccatttccg ctcgccgcag tcgaacgacc
     6301 gagcgtagcg agtcagtgag cgaggaagcg gaatatatcc tgtatcacat attctgctga
     6361 cgcaccggtg cagccttttt tctcctgcca catgaagcac ttcactgaca ccctcatcag
     6421 tgccaacata gtaagccagt atacactccc ctagcgctga tgtccggcgg tgcttttgcc
     6481 gttacgcacc accccgtcag tagctgaaca ggagggacag ctgatagaaa cagaagccac
     6541 tggagcacct caaaaacacc atcatacact aaatcagtaa gttggcagca tcacccgacg
     6601 cactttgcgc cgaataaata cctgtgacgg aagatcactt cgcagaataa ataaatcctg
     6661 gtgtccctgt tgataccggg aagccctggg ccaacttttg gcgaaaatga gacgttgatc
     6721 ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt
     6781 ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     6841 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     6901 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     6961 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     7021 atgctcatcc ggaattc
//
