LOCUS       sequence-290133-        6797 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial Expression of HsPAN3_245-687-M546H.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6797)
  AUTHORS   Christie M, Boland A, Huntzinger E, Weichenrieder O, Izaurralde E
  TITLE     Structure of the PAN3 pseudokinase reveals the basis for 
            interactions with the PAN2 deadenylase and the GW182 proteins.
  JOURNAL   Mol Cell. 2013 Aug 8;51(3):360-73. doi: 
            10.1016/j.molcel.2013.07.011.
  PUBMED    23932717
REFERENCE   2  (bases 1 to 6797)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.molcel.2013.07"; journalName: "Mol Cell"; date: 
            "2013-08-8- 8"; volume: "51"; issue: "3"; pages: "360-73"
COMMENT     Sequence Label: pnYC-NpM-HsPAN3_245-687-M546H_U
FEATURES             Location/Qualifiers
     source          1..6797
                     /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(5029..5047)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5043..5090
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5265..5270,5271..5287,5288..5293))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      5405..5950
                     /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     5851..5870
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6067..6084
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        6476..6578
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(6579..6797,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6645..6664)
                     /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 atggctgatg aactccgaca
     3661 ggagctgatc aacagacatt taataacaat ggctcaaatt gatcaagcag atatgccagc
     3721 agttcctaca gaggttgaca gctaccatag cctattccct ctagaaccac tgccacctcc
     3781 caaccggata cagaaatcaa gtaattttgg atatattaca tcttgctaca aagctgtaaa
     3841 cagcaaagat gatctgccat attgccttcg gaggatacat ggttttcgtc ttgttaacac
     3901 aaagtgcatg gtgttggtcg acatgtggaa gaaaattcaa cactcaaata tcgtaacttt
     3961 gcgtgaagta tttaccacta aagcatttgc tgagccctct cttgtgtttg cacatgattt
     4021 ccatgctgga ggagaaacta tgatgagcag acactttaat gaccctaatg ctgatgccta
     4081 cttcaccaag agaaagtggg gtcagcacga gggaccattg cccaggcagc atgctggatt
     4141 attgccagaa tctcttattt gggcatatat tgtccaacta agttctgcat tgcgtaccat
     4201 tcatacagca ggtttggcat gtcgagttat ggatccaaca aagattctga taactggcaa
     4261 aacaaggttg cgagtaaatt gtgttggagt ttttgatgtt ttaacatttg ataacagtca
     4321 aaataataat ccattggcat taatggccca gtaccagcaa gcagatctga tatcattagg
     4381 aaaagttgtg ttggctttgg cttgcaactc tttggcagga attcagcgag agaatttaca
     4441 gaaagccatg gaactggtga caatcaacta ttcctctgac ctgaagaatc tgattttgta
     4501 tttgttgact gaccaaaaca ggatgcgaag tgtaaatgac atccacccca tgattggtgc
     4561 tcgattttat actcaattgg atgctgctca aatgagaaat gatgtcatag aggaagacct
     4621 tgcaaaggag gttcaaaatg gaagactgtt taggctccta gcaaaattgg gaacaatcaa
     4681 tgagaggccg gagtttcaga aggatcccac ttggtcagag actggagacc gttatctgtt
     4741 gaaactcttt agggatcatc tttttcatca ggtgacagaa gcaggtgctc cctggattga
     4801 cctcagtcat ataatttctt gtcttaacaa gctagatgct ggtgtgccag aaaaaataag
     4861 cctgatttcc agagatgaga agagtgtact tgtggtgacc tacagtgact taaagcgctg
     4921 ctttgaaaat acttttcaag aactgattgc agctgcaaat ggtcagttgt agcctaggag
     4981 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     5041 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     5101 gaactatatc cggatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc
     5161 tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca
     5221 cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat
     5281 gataagctgt caaacatgag aattacaact tatatcgtat ggggctgact tcaggtgcta
     5341 catttgaaga gataaattgc actgaaatca agaaatattt tatctgatta ataagatgat
     5401 cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc tgaaaacgaa aaaaccgcct
     5461 tgcagggcgg tttttcgaag gttctctgag ctaccaactc tttgaaccga ggtaactggc
     5521 ttggaggagc gcagtcacca aaacttgtcc tttcagttta gccttaaccg gcgcatgact
     5581 tcaagactaa ctcctctaaa tcaattacca gtggctgctg ccagtggtgc ttttgcatgt
     5641 ctttccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc ggactgaacg
     5701 gggggttcgt gcatacagtc cagcttggag cgaactgcct acccggaact gagtgtcagg
     5761 cgtggaatga gacaaacgcg gccataacag cggaatgaca ccggtaaacc gaaaggcagg
     5821 aacaggagag cgcacgaggg agccgccagg gggaaacgcc tggtatcttt atagtcctgt
     5881 cgggtttcgc caccactgat ttgagcgtca gatttcgtga tgcttgtcag gggggcggag
     5941 cctatggaaa aacggctttg ccgcggccct ctcacttccc tgttaagtat cttcctggca
     6001 tcttccagga aatctccgcc ccgttcgtaa gccatttccg ctcgccgcag tcgaacgacc
     6061 gagcgtagcg agtcagtgag cgaggaagcg gaatatatcc tgtatcacat attctgctga
     6121 cgcaccggtg cagccttttt tctcctgcca catgaagcac ttcactgaca ccctcatcag
     6181 tgccaacata gtaagccagt atacactccc ctagcgctga tgtccggcgg tgcttttgcc
     6241 gttacgcacc accccgtcag tagctgaaca ggagggacag ctgatagaaa cagaagccac
     6301 tggagcacct caaaaacacc atcatacact aaatcagtaa gttggcagca tcacccgacg
     6361 cactttgcgc cgaataaata cctgtgacgg aagatcactt cgcagaataa ataaatcctg
     6421 gtgtccctgt tgataccggg aagccctggg ccaacttttg gcgaaaatga gacgttgatc
     6481 ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt
     6541 ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     6601 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     6661 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     6721 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     6781 atgctcatcc ggaattc
//
