LOCUS       sequence-290933-        7046 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsNot1iso1_1844-2361-HCAtoEEE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7046)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7046)
  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-HsNot1iso1_1844-2361-HCAtoEEE_AD
FEATURES             Location/Qualifiers
     source          1..7046
                     /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(5278..5296)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5292..5339
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5514..5519,5520..5536,5537..5542))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      5654..6199
                     /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     6100..6119
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6316..6333
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        6725..6827
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(6828..7046,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6894..6913)
                     /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 gaccctccag gcctgaggga
     3661 gaaggcagag tatcttctga gggaatgggt gaatctctac cattcagcag cagctggccg
     3721 cgacagtacc aaagctttct ctgcatttgt tggacagatg caccagcaag gaatactgaa
     3781 gaccgatgat ctcataacaa ggttctttcg tctgtgtact gaaatgtgtg ttgaaatcag
     3841 ttaccgtgct caggctgagc agcagcacaa tcctgctgcc aatcccacca tgatccgagc
     3901 caagtgctat cacaacctgg atgcctttgt tcgactcatt gcactgctcg tgaaacactc
     3961 aggggaggcc accaacactg tcacaaagat taatctgctg aacaaggtcc ttggtatagt
     4021 agtgggagtt ctccttcagg atcatgatgt tcgtcagagt gaatttcagc aacttcccta
     4081 ccatcgaatt tttatcatgc ttctcttgga actcaatgca cctgagcatg tgttggaaac
     4141 cattaatttc cagacactta cagctttctg caatacattc cacatcttga ggcctaccaa
     4201 agctcctggc tttgtatatg cctggcttga actgatttcc catcggatat ttattgcaag
     4261 aatgctggca catacgccac agcagaaggg gtggcctatg tatgcacagc tactgattga
     4321 tttattcaaa tatttagcgc ctttccttag aaatgtggaa ctcaccaaac ctatgcaaat
     4381 cctctacaag ggcactttaa gagtgctgct ggttcttttg catgatttcc cagagttcct
     4441 ttgtgattac cattatgggt tctgtgatgt gatcccacct aattgtatcc agttaagaaa
     4501 tttgatcctg agtgcctttc caagaaacat gaggctcccc gacccattca ctcctaatct
     4561 aaaggtggac atgttgagtg aaattaacat tgctccccgg attctcacca atttcactgg
     4621 agtaatgcca cctcagttca aaaaggattt ggattcctat cttaaaactc gatcaccagt
     4681 cactttcctg tctgatctgc gcagcaacct acaggtatcc aatgaacctg ggaatcgcta
     4741 caacctccag ctcatcaatg cactggtgct ctatgtcggg actcaggcca ttgcgcacat
     4801 ccacaacaag ggcagcacac cttcaatgag caccatcact cactcagcac acatggatat
     4861 cttccagaat ttggctgtgg acttggacac tgagggtcgc tatctctttt tgaatgcaat
     4921 tgcaaatcag ctccggtacc caaatagcca cactcactac ttcagttgca ccatgctgta
     4981 cctttttgca gaggccaata cggaagccat ccaagaacag atcacaagag ttctcttgga
     5041 acggttgatt gtaaataggc cacatccttg gggtcttctt attaccttca ttgagctgat
     5101 taaaaaccca gcgtttaagt tctggaacca tgaatttgta gaggaggaac cagaaatcga
     5161 aaagttattc cagtcggtcg cacagtgctg catgtaagga tcctaatagt ctagagctag
     5221 ccctaggaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     5281 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     5341 tgaaaggagg aactatatcc ggatatcccg caagaggccc ggcagtaccg gcataaccaa
     5401 gcctatgcct acagcatcca gggtgacggt gccgaggatg acgatgagcg cattgttaga
     5461 tttcatacac ggtgcctgac tgcgttagca atttaactgt gataaactac cgcattaaag
     5521 cttatcgatg ataagctgtc aaacatgaga attacaactt atatcgtatg gggctgactt
     5581 caggtgctac atttgaagag ataaattgca ctgaaatcaa gaaatatttt atctgattaa
     5641 taagatgatc ttcttgagat cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa
     5701 aaaccgcctt gcagggcggt ttttcgaagg ttctctgagc taccaactct ttgaaccgag
     5761 gtaactggct tggaggagcg cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg
     5821 cgcatgactt caagactaac tcctctaaat caattaccag tggctgctgc cagtggtgct
     5881 tttgcatgtc tttccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5941 gactgaacgg ggggttcgtg catacagtcc agcttggagc gaactgccta cccggaactg
     6001 agtgtcaggc gtggaatgag acaaacgcgg ccataacagc ggaatgacac cggtaaaccg
     6061 aaaggcagga acaggagagc gcacgaggga gccgccaggg ggaaacgcct ggtatcttta
     6121 tagtcctgtc gggtttcgcc accactgatt tgagcgtcag atttcgtgat gcttgtcagg
     6181 ggggcggagc ctatggaaaa acggctttgc cgcggccctc tcacttccct gttaagtatc
     6241 ttcctggcat cttccaggaa atctccgccc cgttcgtaag ccatttccgc tcgccgcagt
     6301 cgaacgaccg agcgtagcga gtcagtgagc gaggaagcgg aatatatcct gtatcacata
     6361 ttctgctgac gcaccggtgc agcctttttt ctcctgccac atgaagcact tcactgacac
     6421 cctcatcagt gccaacatag taagccagta tacactcccc tagcgctgat gtccggcggt
     6481 gcttttgccg ttacgcacca ccccgtcagt agctgaacag gagggacagc tgatagaaac
     6541 agaagccact ggagcacctc aaaaacacca tcatacacta aatcagtaag ttggcagcat
     6601 cacccgacgc actttgcgcc gaataaatac ctgtgacgga agatcacttc gcagaataaa
     6661 taaatcctgg tgtccctgtt gataccggga agccctgggc caacttttgg cgaaaatgag
     6721 acgttgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg
     6781 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
     6841 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     6901 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     6961 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     7021 gcctgatgaa tgctcatccg gaattc
//
