LOCUS       sequence-355322-        5570 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmXRN1_1323-1355.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5570)
  AUTHORS   Braun JE, Truffault V, Boland A, Huntzinger E, Chang CT, Haas G, 
            Weichenrieder O, Coles M, Izaurralde E
  TITLE     A direct interaction between DCP1 and XRN1 couples mRNA decapping to
            5' exonucleolytic degradation.
  JOURNAL   Nat Struct Mol Biol. 2012 Dec;19(12):1324-31. doi: 
            10.1038/nsmb.2413. Epub 2012 Nov 11.
  PUBMED    23142987
REFERENCE   2  (bases 1 to 5570)
  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.2413";
            journalName: "Nat Struct Mol Biol"; date: "2012-12"; volume: "19"; 
            issue: "12"; pages: "1324-31"
COMMENT     Sequence Label: pnYC-NvHN-DmXRN1_1323-1355_N
FEATURES             Location/Qualifiers
     source          1..5570
                     /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             2060..2077
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2093..3577
                     /codon_start=1
                     /gene="nusA"
                     /product="transcription elongation protein (N utilization 
                     substance protein A)"
                     /label=NusA
                     /note="highly soluble in E. coli"
                     /translation="MDKEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDV
                     RVQIDRKSGDFDTFRRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIESVTFDR
                     ITTQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNISLDLGNNAEAVIL
                     REDMLPRENFRPGDRVRGVLYSVRPEARGAQLFVTRSKPEMLIELFRIEVPEIGEEVIE
                     IKAAARDPGSRAKIAVKTNDKRIDPVGACVGMRGARVQAVSTELGGERIDIVLWDDNPA
                     QFVINAMAPADVASIVVDEDKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMT
                     VDDLQAKHQAEAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEIEGLD
                     EPTVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAARGVCTLED
                     LAEQGIDDLADIEGLTDEKAGALIMAARNICWFGDEA"
     CDS             3593..3613
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3802..3820)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3816..3863
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4038..4043,4044..4060,4061..4066))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      4178..4723
                     /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     4624..4643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4840..4857
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5249..5351
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(5352..5570,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5418..5437)
                     /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 agcggcagcc atcatcatca tcatcacagc ggcagcggct ccatggacaa
     2101 agaaattttg gctgtagttg aagccgtatc caatgaaaag gcgctacctc gcgagaagat
     2161 tttcgaagca ttggaaagcg ccctggcgac agcaacaaag aaaaaatatg aacaagagat
     2221 cgacgtccgc gtacagatcg atcgcaaaag cggtgatttt gacactttcc gtcgctggtt
     2281 agttgttgat gaagtcaccc agccgaccaa ggaaatcacc cttgaagccg cacgttatga
     2341 agatgaaagc ctgaacctgg gcgattacgt tgaagatcag attgagtctg ttacctttga
     2401 ccgtatcact acccagacgg caaaacaggt tatcgtgcag aaagtgcgtg aagccgaacg
     2461 tgcgatggtg gttgatcagt tccgtgaaca cgaaggtgaa atcatcaccg gcgtggtgaa
     2521 aaaagtaaac cgcgacaaca tctctctgga tctgggcaac aacgctgaag ccgtgatcct
     2581 gcgcgaagat atgctgccgc gtgaaaactt ccgccctggc gaccgcgttc gtggcgtgct
     2641 ctattccgtt cgcccggaag cgcgtggcgc ccaactgttc gtcactcgtt ccaagccgga
     2701 aatgctgatc gaactgttcc gtattgaagt gccagaaatc ggcgaagaag tgattgaaat
     2761 taaagcagcg gctcgcgatc cgggttctcg tgcgaaaatc gcggtgaaaa ccaacgataa
     2821 acgtatcgat ccggtaggtg cttgcgtagg tatgcgtggc gcccgtgttc aggcggtgtc
     2881 tactgaactg ggtggcgagc gtatcgatat cgtcctgtgg gatgataacc cggcgcagtt
     2941 cgtgattaac gcaatggcac cggcagacgt tgcttctatc gtggtggatg aagataaaca
     3001 cacgatggac atcgccgttg aagccggtaa tctggcgcag gcgattggcc gtaacggtca
     3061 gaacgtgcgt ctggcttcgc aactgagcgg ttgggaactc aacgtgatga ccgttgacga
     3121 cctgcaagct aagcatcagg cggaagcgca cgcagcgatc gacaccttca ccaaatatct
     3181 cgacatcgac gaagacttcg cgactgttct ggtagaagaa ggcttctcga cgctggaaga
     3241 attggcctat gtgccgatga aagagctgtt ggaaatcgaa ggccttgatg agccgaccgt
     3301 tgaagcactg cgcgagcgtg ctaaaaatgc actggccacc attgcacagg cccaggaaga
     3361 aagcctcggt gataacaaac cggctgacga tctgctgaac cttgaagggg tagatcgtga
     3421 tttggcattc aaactggccg cccgtggcgt ttgtacgctg gaagacctcg ccgaacaggg
     3481 cattgatgat ctggctgata tcgaagggtt gaccgacgaa aaagccggag cactgattat
     3541 ggctgcccgt aatatttgct ggttcggtga cgaagcgggt accggcagcg gcgaaaacct
     3601 ttacttccag ggccatatgc tacttcaaca gcaaagagct ccgtttcccg gacaaatgcc
     3661 caatctaccc aagccaccgc tcttctggca gcaagaggcg cagaagcagg aggcgctgta
     3721 gggatcctaa tagtctagag ctagccctag gagatccggc tgctaacaaa gcccgaaagg
     3781 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     3841 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatat cccgcaagag
     3901 gcccggcagt accggcataa ccaagcctat gcctacagca tccagggtga cggtgccgag
     3961 gatgacgatg agcgcattgt tagatttcat acacggtgcc tgactgcgtt agcaatttaa
     4021 ctgtgataaa ctaccgcatt aaagcttatc gatgataagc tgtcaaacat gagaattaca
     4081 acttatatcg tatggggctg acttcaggtg ctacatttga agagataaat tgcactgaaa
     4141 tcaagaaata ttttatctga ttaataagat gatcttcttg agatcgtttt ggtctgcgcg
     4201 taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct
     4261 gagctaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca ccaaaacttg
     4321 tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct aaatcaatta
     4381 ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc aagacgatag
     4441 ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca gtccagcttg
     4501 gagcgaactg cctacccgga actgagtgtc aggcgtggaa tgagacaaac gcggccataa
     4561 cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga gggagccgcc
     4621 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact gatttgagcg
     4681 tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct ttgccgcggc
     4741 cctctcactt ccctgttaag tatcttcctg gcatcttcca ggaaatctcc gccccgttcg
     4801 taagccattt ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa
     4861 gcggaatata tcctgtatca catattctgc tgacgcaccg gtgcagcctt ttttctcctg
     4921 ccacatgaag cacttcactg acaccctcat cagtgccaac atagtaagcc agtatacact
     4981 cccctagcgc tgatgtccgg cggtgctttt gccgttacgc accaccccgt cagtagctga
     5041 acaggaggga cagctgatag aaacagaagc cactggagca cctcaaaaac accatcatac
     5101 actaaatcag taagttggca gcatcacccg acgcactttg cgccgaataa atacctgtga
     5161 cggaagatca cttcgcagaa taaataaatc ctggtgtccc tgttgatacc gggaagccct
     5221 gggccaactt ttggcgaaaa tgagacgttg atcggcacgt aagaggttcc aactttcacc
     5281 ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta
     5341 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
     5401 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
     5461 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     5521 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc
//
