LOCUS       sequence-357350-        6356 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsPAN3_245-687-M546FP547G.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6356)
  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 6356)
  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: 
            "10.1016/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-M546FP547G_U
FEATURES             Location/Qualifiers
     source          1..6356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1853..1872
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1961..1980
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1961..1979
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1980..2004
                     /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             2019..2041
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2048..3148
                     /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             3164..3187
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(4588..4606)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4602..4649
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4824..4829,4830..4846,4847..4852))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      4964..5509
                     /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     5410..5429
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5626..5643
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        6035..6137
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(6138..6356,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6204..6223)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 cgtatggcaa 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 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
      421 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt
      481 tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgaa agcaaattcg
      541 acccggtcgt cggttcaggg cagggtcgtt aaatagccgc ttatgtctat tgctggttta
      601 ccggtttatt gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc tgaggccagt
      661 ttgctcaggc tctccccgtg gaggtaataa ttgacgatat gatcatttat tctgcctccc
      721 agagcctgat aaaaacggtt agcgcttcgt taatacagat gtaggtgttc cacagggtag
      781 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctt gtttcggcgt
      841 gggtatggtg gcaggccccg tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg
      901 ctgctggaga tggcggacgc gatggatatg ttctgccaag ggttggtttg cgcattcaca
      961 gttctccgca agaattgatt ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc
     1021 cgccggcttc cattcaggtc gaggtggccc ggctccatgc accgcgacgc aacgcgggga
     1081 ggcagacaag gtatagggcg gcgcctacaa tccatgccaa cccgttccat gtgctcgccg
     1141 aggcggcata aatcgccgtg acgatcagcg gtccaatgat cgaagttagg ctggtaagag
     1201 ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg
     1261 cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca
     1321 tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg
     1381 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
     1441 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
     1501 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
     1561 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
     1621 agctgactgg gttgaaggct ctcaagggca tcggtcgacg ctctccctta tgcgactcct
     1681 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     1741 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     1801 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     1861 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     1921 cggcgtagag gatcagacta agatctcgat cccgcgaaat taatacgact cactataggg
     1981 gaattgtgag cggataacaa ttccccacta gtaataattt tgtttaactt taagaaggag
     2041 atataccatg gaaactgaag aaggtaaact ggtaatctgg attaacggcg ataaaggcta
     2101 taacggtctc gctgaagtcg gtaagaaatt cgagaaagat accggaatta aagtcaccgt
     2161 tgagcatccg gataaactgg aagagaaatt cccacaggtt gcggcaactg gcgatggccc
     2221 tgacattatc ttctgggcac acgaccgctt tggtggctac gctcaatctg gcctgttggc
     2281 tgaaatcacc ccggacaaag cgttccagga caagctgtat ccgtttacct gggatgccgt
     2341 acgttacaac ggcaagctga ttgcttaccc gatcgctgtt gaagcgttat cgctgattta
     2401 taacaaagac ctgctgccga acccgccaaa aacctgggaa gagatcccgg cgctggataa
     2461 agaactgaaa gcgaaaggta agagcgccct gatgttcaac ctgcaagaac cgtacttcac
     2521 ctggccgctg attgctgctg acgggggtta tgcgttcaag tatgaaaacg gcaagtacga
     2581 cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg ggtctgacct tcctggttga
     2641 cctgattaaa aacaaacaca tgaatgcaga caccgattac tccatcgcag aagctgcctt
     2701 taataaaggc gaaacagcga tgaccatcaa cggcccgtgg gcatggtcca acatcgacac
     2761 cagcaaagtg aattatggtg taacggtact gccgaccttc aagggtcaac catccaaacc
     2821 gttcgttggc gtgctgagcg caggtattaa cgccgccagt ccgaacaaag agctggcgaa
     2881 agagttcctc gaaaactatc tgctgactga tgaaggtctg gaagcggtta ataaagacaa
     2941 accgctgggt gccgtagcgc tgaagtctta cgaggaagag ttggcgaaag atccacgtat
     3001 tgccgccacg atggaaaacg cccagaaagg tgaaatcatg ccgaacatcc cgcagatgtc
     3061 cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc gccagcggtc gtcagactgt
     3121 cgatgaagcc cttaaagacg cgcagactgg taccggcagc ggcctggaag ttctgttcca
     3181 ggggccccat atgctcgaga tggctgatga actccgacag gagctgatca acagacattt
     3241 aataacaatg gctcaaattg atcaagcaga tatgccagca gttcctacag aggttgacag
     3301 ctaccatagc ctattccctc tagaaccact gccacctccc aaccggatac agaaatcaag
     3361 taattttgga tatattacat cttgctacaa agctgtaaac agcaaagatg atctgccata
     3421 ttgccttcgg aggatacatg gttttcgtct tgttaacaca aagtgcatgg tgttggtcga
     3481 catgtggaag aaaattcaac actcaaatat cgtaactttg cgtgaagtat ttaccactaa
     3541 agcatttgct gagccctctc ttgtgtttgc acatgatttc catgctggag gagaaactat
     3601 gatgagcaga cactttaatg accctaatgc tgatgcctac ttcaccaaga gaaagtgggg
     3661 tcagcacgag ggaccattgc ccaggcagca tgctggatta ttgccagaat ctcttatttg
     3721 ggcatatatt gtccaactaa gttctgcatt gcgtaccatt catacagcag gtttggcatg
     3781 tcgagttatg gatccaacaa agattctgat aactggcaaa acaaggttgc gagtaaattg
     3841 tgttggagtt tttgatgttt taacatttga taacagtcaa aataataatc cattggcatt
     3901 aatggcccag taccagcaag cagatctgat atcattagga aaagttgtgt tggctttggc
     3961 ttgcaactct ttggcaggaa ttcagcgaga gaatttacag aaagccatgg aactggtgac
     4021 aatcaactat tcctctgacc tgaagaatct gattttgtat ttgttgactg accaaaacag
     4081 gatgcgaagt gtaaatgaca tcttcggcat gattggtgct cgattttata ctcaattgga
     4141 tgctgctcaa atgagaaatg atgtcataga ggaagacctt gcaaaggagg ttcaaaatgg
     4201 aagactgttt aggctcctag caaaattggg aacaatcaat gagaggccgg agtttcagaa
     4261 ggatcccact tggtcagaga ctggagaccg ttatctgttg aaactcttta gggatcatct
     4321 ttttcatcag gtgacagaag caggtgctcc ctggattgac ctcagtcata taatttcttg
     4381 tcttaacaag ctagatgctg gtgtgccaga aaaaataagc ctgatttcca gagatgagaa
     4441 gagtgtactt gtggtgacct acagtgactt aaagcgctgc tttgaaaata cttttcaaga
     4501 actgattgca gctgcaaatg gtcagttgta gcctaggaga tccggctgct aacaaagccc
     4561 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     4621 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatatcccg
     4681 caagaggccc ggcagtaccg gcataaccaa gcctatgcct acagcatcca gggtgacggt
     4741 gccgaggatg acgatgagcg cattgttaga tttcatacac ggtgcctgac tgcgttagca
     4801 atttaactgt gataaactac cgcattaaag cttatcgatg ataagctgtc aaacatgaga
     4861 attacaactt atatcgtatg gggctgactt caggtgctac atttgaagag ataaattgca
     4921 ctgaaatcaa gaaatatttt atctgattaa taagatgatc ttcttgagat cgttttggtc
     4981 tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt ttttcgaagg
     5041 ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg cagtcaccaa
     5101 aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac tcctctaaat
     5161 caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt ggactcaaga
     5221 cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg catacagtcc
     5281 agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag acaaacgcgg
     5341 ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc gcacgaggga
     5401 gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc accactgatt
     5461 tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa acggctttgc
     5521 cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa atctccgccc
     5581 cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga gtcagtgagc
     5641 gaggaagcgg aatatatcct gtatcacata ttctgctgac gcaccggtgc agcctttttt
     5701 ctcctgccac atgaagcact tcactgacac cctcatcagt gccaacatag taagccagta
     5761 tacactcccc tagcgctgat gtccggcggt gcttttgccg ttacgcacca ccccgtcagt
     5821 agctgaacag gagggacagc tgatagaaac agaagccact ggagcacctc aaaaacacca
     5881 tcatacacta aatcagtaag ttggcagcat cacccgacgc actttgcgcc gaataaatac
     5941 ctgtgacgga agatcacttc gcagaataaa taaatcctgg tgtccctgtt gataccggga
     6001 agccctgggc caacttttgg cgaaaatgag acgttgatcg gcacgtaaga ggttccaact
     6061 ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag
     6121 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
     6181 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     6241 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     6301 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattc
//
