LOCUS       sequence-357797-        6194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of NcPAN2_1-390.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6194)
  AUTHORS   Jonas S, Christie M, Peter D, Bhandari D, Loh B, Huntzinger E, 
            Weichenrieder O, Izaurralde E
  TITLE     An asymmetric PAN3 dimer recruits a single PAN2 exonuclease to 
            mediate mRNA deadenylation and decay.
  JOURNAL   Nat Struct Mol Biol. 2014 Jul;21(7):599-608. doi: 10.1038/nsmb.2837.
            Epub 2014 Jun 1.
  PUBMED    24880343
REFERENCE   2  (bases 1 to 6194)
  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: "10.1038/nsmb.2837";
            journalName: "Nat Struct Mol Biol"; date: "2014-07"; volume: "21"; 
            issue: "7"; pages: "599-608"
COMMENT     Sequence Label: pnYC-NpM-NcPAN2_1-390_Y
FEATURES             Location/Qualifiers
     source          1..6194
                     /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(4426..4444)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4440..4487
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4662..4667,4668..4684,4685..4690))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      4802..5347
                     /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     5248..5267
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5464..5481
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5873..5975
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(5976..6194,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6042..6061)
                     /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 atggattcgc gggattggac ccagctgggc tgtgttgcct acccgtcacc
     3241 aatccatcca gactaccatg ccggacccgc gtcgacaata gccttcgaca accaagatga
     3301 gctgctatgg atcggcaccc aaaagggctt tgccggctct ttcatcggcc gcgagctcaa
     3361 gcgattcact gccttccgaa tacaccccga aaccgacggt cccctccgcc aattcctctt
     3421 cgtcgacaag ggcgtcatct tcctcggaag ccggagcgtc tacatggcag cacgtagtgg
     3481 cgtgcccatc tggagtatcc gccatgagtc catgcaggat ctgcgtgcca tgtccttcac
     3541 cagcaagggg acctccgaga tcctcgtggc cggctggcag aacaagatgt tggtgattga
     3601 cgtcaacaag ggtgaggtcg tcaaggagct cccgacacaa gatcagtact ccttcctcaa
     3661 gatgagccgg tacatttgcg cggcaacgaa caagggcacc gtcaacatac tggaccctat
     3721 cacttttact atcaagaaac aatggcaggc ccacggggct ttcatcaacg atctggacac
     3781 gtccaacgac tttattgtca cctgcggtgg ctcacacagg cagacacaca acaccccggc
     3841 catactcgac ccttacgtca aggtgtttga cctgaagaac atgagtgcta tgaacccagt
     3901 tccctttgcg ccgctggccg ctcacgtgag gatgcacccg aggatgttga caacggccat
     3961 tgtcgtcaac caggcgggcc agatccacgt gacggatctg ttgaatccaa gcaacagcca
     4021 ggtctgctac acgcagcctc agggcgtggt gctacatttc gatgtgtcga gaaccggaga
     4081 aggaaaagcg cttgcggata acaagcacaa cacctacgtc tggggatcgc ccaacaagat
     4141 ccagtttacc gagataggaa tcccgcctcg gttgccggac cctccccagc cgtcattgtt
     4201 gccgccagat ccagacatgt tggaggaatt acccctgagc cgcatcggct tgccttatta
     4261 ccgagaacag ctcttctcag ccttgccgcc ggatatcatc tccgatgtgg gcgctcctcc
     4321 gcagcagatc gatccgaaca tcttgagcac acttaccaag tgagctagcc ctaggagatc
     4381 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
     4441 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
     4501 ctatatccgg atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac
     4561 agcatccagg gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg
     4621 tgcctgactg cgttagcaat ttaactgtga taaactaccg cattaaagct tatcgatgat
     4681 aagctgtcaa acatgagaat tacaacttat atcgtatggg gctgacttca ggtgctacat
     4741 ttgaagagat aaattgcact gaaatcaaga aatattttat ctgattaata agatgatctt
     4801 cttgagatcg ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa accgccttgc
     4861 agggcggttt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt aactggcttg
     4921 gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg catgacttca
     4981 agactaactc ctctaaatca attaccagtg gctgctgcca gtggtgcttt tgcatgtctt
     5041 tccgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga ctgaacgggg
     5101 ggttcgtgca tacagtccag cttggagcga actgcctacc cggaactgag tgtcaggcgt
     5161 ggaatgagac aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac
     5221 aggagagcgc acgagggagc cgccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5281 gtttcgccac cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct
     5341 atggaaaaac ggctttgccg cggccctctc acttccctgt taagtatctt cctggcatct
     5401 tccaggaaat ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag
     5461 cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt atcacatatt ctgctgacgc
     5521 accggtgcag ccttttttct cctgccacat gaagcacttc actgacaccc tcatcagtgc
     5581 caacatagta agccagtata cactccccta gcgctgatgt ccggcggtgc ttttgccgtt
     5641 acgcaccacc ccgtcagtag ctgaacagga gggacagctg atagaaacag aagccactgg
     5701 agcacctcaa aaacaccatc atacactaaa tcagtaagtt ggcagcatca cccgacgcac
     5761 tttgcgccga ataaatacct gtgacggaag atcacttcgc agaataaata aatcctggtg
     5821 tccctgttga taccgggaag ccctgggcca acttttggcg aaaatgagac gttgatcggc
     5881 acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc gtattttttg
     5941 agttatcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata
     6001 ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg
     6061 ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa
     6121 agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg
     6181 ctcatccgga attc
//
