LOCUS       sequence-290236-        6692 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial Expression of CtPAN3opt_171-566.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6692)
  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 6692)
  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: "10.1038/nsmb.2837";
            journalName: "Nat Struct Mol Biol"; date: "2014-07"; volume: "21"; 
            issue: "7"; pages: "599-608"
COMMENT     Sequence Label: pnYC-NpM-CtPAN3opt_171-566_V
FEATURES             Location/Qualifiers
     source          1..6692
                     /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"
     terminator      4938..4985
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5160..5165,5166..5182,5183..5188))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      5300..5845
                     /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     5746..5765
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5962..5979
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        6371..6473
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(6474..6692,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6540..6559)
                     /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 cttaagcaat tgcactatca
     3661 tctgtatcag ccgtttggtc cgtatcgtca agaactgcag ccgtggcagc gtgcaaccta
     3721 tgattttttc atgccgcaga atctgcgtga agatctgcag aaaaaacagt ttgcaaccct
     3781 gcaggttatt ccgagctatc gtctgaccaa tgaaaaagcc attctgaacg ttatgaaaga
     3841 ttggaaaaaa atcaaaaacg ccagcatcgt gaccatccat gaagttttta ccacccgtga
     3901 atttggtgat agcagcctga tttttgcgta tgattttcat ccgctgagca aaaccctgca
     3961 agaacatcat tttcagccga ttcatggtaa tcgttatcgt ccgcctccgg cagttccgga
     4021 aaacaccatt tggggttata tttgtcagat tgccaatgcc ctgaaaacca ttcatagcaa
     4081 tcgtctggca gcacgttgtc tggaaccgag caaaattatc ctgaccgata ttaatcgtat
     4141 tcgtctgagc gcatgtgcaa ttctggatgt tgttcagttt ggtatgaata gccgtagcgt
     4201 tgttgagctg cagcaagaag attttgttaa attcggcaaa ctgattctga gcctggcaac
     4261 cggcaccctg cctgcacatc tgaataacat tccggcagca ctggaaaccc tgggtaacaa
     4321 atattcagca aatctgaaaa gcgcagtgaa ttggctgctg gataccagct caggtgaaac
     4381 caaaaccatt gaacatttta tgaccggtat tgcaagccag atgaccacct tttttgatct
     4441 ggcactgcag gataatgatg agaaactgtt tcatctggca cgcgaagttg aaaatggtcg
     4501 tattgcacgt agcctgatga aactgctgac cattctggaa cgcggtgatt atgacggtgt
     4561 tccgagctgg tcagaaaccg gtgatcgtta tcagctgaaa ctgttccgtg attatgtttt
     4621 tcatcgtgtt gatgcagatg gcaaaccgaa tctgagtatt ggtcatatgc tgacctgtat
     4681 gagcaaactg gaagccggtg ttgatgaaaa cattctgctg acctcacgtg ataatgaaac
     4741 cgtttttgtt ctgagctatc gcgaactgcg tcagatgtat gatcgtgcat ttaatgaact
     4801 ggtgaaagca agcaaaacag gtgcaccggg tgcaaatacc taaggatcct aatagtctag
     4861 agctagccct aggagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     4921 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     4981 ttttgctgaa aggaggaact atatccggat atcccgcaag aggcccggca gtaccggcat
     5041 aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt
     5101 gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca
     5161 ttaaagctta tcgatgataa gctgtcaaac atgagaatta caacttatat cgtatggggc
     5221 tgacttcagg tgctacattt gaagagataa attgcactga aatcaagaaa tattttatct
     5281 gattaataag atgatcttct tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa
     5341 acgaaaaaac cgccttgcag ggcggttttt cgaaggttct ctgagctacc aactctttga
     5401 accgaggtaa ctggcttgga ggagcgcagt caccaaaact tgtcctttca gtttagcctt
     5461 aaccggcgca tgacttcaag actaactcct ctaaatcaat taccagtggc tgctgccagt
     5521 ggtgcttttg catgtctttc cgggttggac tcaagacgat agttaccgga taaggcgcag
     5581 cggtcggact gaacgggggg ttcgtgcata cagtccagct tggagcgaac tgcctacccg
     5641 gaactgagtg tcaggcgtgg aatgagacaa acgcggccat aacagcggaa tgacaccggt
     5701 aaaccgaaag gcaggaacag gagagcgcac gagggagccg ccagggggaa acgcctggta
     5761 tctttatagt cctgtcgggt ttcgccacca ctgatttgag cgtcagattt cgtgatgctt
     5821 gtcagggggg cggagcctat ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta
     5881 agtatcttcc tggcatcttc caggaaatct ccgccccgtt cgtaagccat ttccgctcgc
     5941 cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg aagcggaata tatcctgtat
     6001 cacatattct gctgacgcac cggtgcagcc ttttttctcc tgccacatga agcacttcac
     6061 tgacaccctc atcagtgcca acatagtaag ccagtataca ctcccctagc gctgatgtcc
     6121 ggcggtgctt ttgccgttac gcaccacccc gtcagtagct gaacaggagg gacagctgat
     6181 agaaacagaa gccactggag cacctcaaaa acaccatcat acactaaatc agtaagttgg
     6241 cagcatcacc cgacgcactt tgcgccgaat aaatacctgt gacggaagat cacttcgcag
     6301 aataaataaa tcctggtgtc cctgttgata ccgggaagcc ctgggccaac ttttggcgaa
     6361 aatgagacgt tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac
     6421 taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga
     6481 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
     6541 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
     6601 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
     6661 ttgcccgcct gatgaatgct catccggaat tc
//
