LOCUS       sequence-357208-        6353 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmPAN3_349-790-P652G.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6353)
  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 6353)
  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-DmPAN3_349-790-P652G_T
FEATURES             Location/Qualifiers
     source          1..6353
                     /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(4585..4603)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4599..4646
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4821..4826,4827..4843,4844..4849))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      4961..5506
                     /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     5407..5426
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5623..5640
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        6032..6134
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(6135..6353,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6201..6220)
                     /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 tgcccgacga gatgcgcgcc gaagtcctcg cccgcaacga
     3241 gatctctaac ctgatcatgg acgcagcgga ggctgcgcag cacgctctac cgctggaggt
     3301 ggagaactac catgccctgt atccactgga accgccggcg cagccgttgc acgccaagct
     3361 cacgttcccc gccaccactt atagggccac gcacaacacg acgggctaca agtactgcct
     3421 gcgaagaata catggtttcc gcctgcagtc gaccaagtgc atgacgctgg tggagatgtg
     3481 gaagaagctg cagcacacga atgtggtaca actgcgtgag gtgttcacga cgaaagcatt
     3541 tggtgataac tctttagtat tagtgtacga ctatcatccc ggctcacaaa cattactggc
     3601 caaatacttc actccggcgc cagagaccaa cgggtacact gatccattcc aaggcgaggc
     3661 ccgccccttc agtcataaga gtaacatgca gcgaacgagc aatggaccgc tgctaccgga
     3721 ggccaccatc tggtcgatca tcatgcagct gaccgccggc ttaaaagcca tccaccatgc
     3781 cggtcttgcc tgcaaggttc tggatcccac aaagattata gtgacgggca agcgtgtacg
     3841 gtttagctca tgctgcattt cggatataac gcaattcgat ccgaatgcat ccaatccctt
     3901 ggcgctggtt aatatgcatc agcaggacga tctgaccgct ttgggtcgct tagtattagc
     3961 tttggcctgc cgctgtcttc aatccgtgca acgcgacaat gtccagtcca gcatcgatat
     4021 ggtcacgcga aactactcca ccgatctgcg aaactttatt gtttacctct tcaccaccaa
     4081 taatcgtcgc tccgtgaccg acctgatggg gatgattggt gcccgcttct atacgcaact
     4141 ggatgcactg cagagcaaaa ttgatatgca ggaggacgag ttggccaagg agatggagaa
     4201 cgggcggtta tatcgcattt tggtcaaact taatagcatc aacgagcgac ccgactttaa
     4261 tctggactgc acttggtctg agactggtga tagatacatg ttgaaattat tccgtgatta
     4321 tttgttccat tccgtcactg aggatggccg gccctggcta gatcacgcac acattgtaca
     4381 atgcctcaac aagctggatg ctggctccat agagcgcgtg caactgatgt cccgcgacga
     4441 gcaatcggta ctcatcgtct cctatgctga actcaagaat tgtctggaga acgccttctc
     4501 cgaactgatg tcaagtgcgg ccaactgacc taggagatcc ggctgctaac aaagcccgaa
     4561 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     4621 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga tatcccgcaa
     4681 gaggcccggc agtaccggca taaccaagcc tatgcctaca gcatccaggg tgacggtgcc
     4741 gaggatgacg atgagcgcat tgttagattt catacacggt gcctgactgc gttagcaatt
     4801 taactgtgat aaactaccgc attaaagctt atcgatgata agctgtcaaa catgagaatt
     4861 acaacttata tcgtatgggg ctgacttcag gtgctacatt tgaagagata aattgcactg
     4921 aaatcaagaa atattttatc tgattaataa gatgatcttc ttgagatcgt tttggtctgc
     4981 gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc
     5041 tctgagctac caactctttg aaccgaggta actggcttgg aggagcgcag tcaccaaaac
     5101 ttgtcctttc agtttagcct taaccggcgc atgacttcaa gactaactcc tctaaatcaa
     5161 ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt ccgggttgga ctcaagacga
     5221 tagttaccgg ataaggcgca gcggtcggac tgaacggggg gttcgtgcat acagtccagc
     5281 ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca
     5341 taacagcgga atgacaccgg taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc
     5401 gccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc actgatttga
     5461 gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta tggaaaaacg gctttgccgc
     5521 ggccctctca cttccctgtt aagtatcttc ctggcatctt ccaggaaatc tccgccccgt
     5581 tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag
     5641 gaagcggaat atatcctgta tcacatattc tgctgacgca ccggtgcagc cttttttctc
     5701 ctgccacatg aagcacttca ctgacaccct catcagtgcc aacatagtaa gccagtatac
     5761 actcccctag cgctgatgtc cggcggtgct tttgccgtta cgcaccaccc cgtcagtagc
     5821 tgaacaggag ggacagctga tagaaacaga agccactgga gcacctcaaa aacaccatca
     5881 tacactaaat cagtaagttg gcagcatcac ccgacgcact ttgcgccgaa taaatacctg
     5941 tgacggaaga tcacttcgca gaataaataa atcctggtgt ccctgttgat accgggaagc
     6001 cctgggccaa cttttggcga aaatgagacg ttgatcggca cgtaagaggt tccaactttc
     6061 accataatga aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag
     6121 ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat
     6181 ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga
     6241 ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt
     6301 atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttc
//
