LOCUS       sequence-410781-        6074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of CtPAN3_448-555.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6074)
  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 6074)
  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: pnEA-NvH-CtPAN3_448-555_Y
FEATURES             Location/Qualifiers
     source          1..6074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             139..159
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(576..594)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      590..637
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(812..817,818..834,835..840))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(867..885)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        953..1057
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1058..1126,1127..1918)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1276..1295)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2089..2677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2578..2597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2831..2848
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2863..3002
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2928..2947)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3088..3110
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3104..3295)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5650..5669
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5689..5766)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5972..5991
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttccccacta gtaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggtaccg gcagcggcga aaacctttac ttccagggcc atatgctcga ggttgaaaat
      181 ggtcgtattg cacgtagcct gatgaaactg ctgaccattc tggaacgcgg tgattatgac
      241 ggtgttccga gctggtcaga aaccggtgat cgttatcagc tgaaactgtt ccgtgattat
      301 gtttttcatc gtgttgatgc agatggcaaa ccgaatctga gtattggtca tatgctgacc
      361 tgtatgagca aactggaagc cggtgttgat gaaaacattc tgctgacctc acgtgataat
      421 gaaaccgttt ttgttctgag ctatcgcgaa ctgcgtcaga tgtatgatcg tgcatttaat
      481 gaactggtga aatgaggatc ctaatagtct agagctagcc ctaggagatc cggctgctaa
      541 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
      601 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
      661 atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg
      721 gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg
      781 cgttagcaat ttaactgtga taaactaccg cattaaagct tatcgatgat aagctgtcaa
      841 acatgagaat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
      901 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
      961 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     1021 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     1081 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     1141 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1201 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1261 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     1321 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     1381 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     1441 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     1501 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1561 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     1621 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1681 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     1741 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     1801 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     1861 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1921 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1981 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     2041 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2101 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2161 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2221 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2281 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2341 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2401 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2461 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2521 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2581 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2641 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2701 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2761 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2821 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     2881 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     2941 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     3001 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3061 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3121 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     3181 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     3241 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     3301 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     3361 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     3421 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     3481 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     3541 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     3601 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     3661 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     3721 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc
     3781 ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag
     3841 ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg
     3901 tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc
     3961 accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa
     4021 cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccaatgat
     4081 cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc
     4141 tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag
     4201 aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag
     4261 cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     4321 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     4381 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     4441 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     4501 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     4561 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     4621 ccagtcggga aactgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4681 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     4741 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     4801 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     4861 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     4921 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     4981 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     5041 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     5101 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     5161 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     5221 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     5281 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     5341 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     5401 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     5461 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     5521 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     5581 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     5641 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     5701 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     5761 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     5821 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     5881 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     5941 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     6001 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
     6061 cgatcccgcg aaat
//
