LOCUS       sequence-289500-        6095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmXRN1_1323-1383.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6095)
  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 6095)
  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_1323-1383_N
FEATURES             Location/Qualifiers
     source          1..6095
                     /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             2501..2518
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2534..4018
                     /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             4034..4054
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(4327..4345)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4341..4388
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4563..4568,4569..4585,4586..4591))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      4703..5248
                     /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     5149..5168
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5365..5382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5774..5876
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(5877..6095,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFGMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5943..5962)
                     /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 gagcggcagc catcatcatc atcatcacag
     2521 cggcagcggc tccatggaca aagaaatttt ggctgtagtt gaagccgtat ccaatgaaaa
     2581 ggcgctacct cgcgagaaga ttttcgaagc attggaaagc gccctggcga cagcaacaaa
     2641 gaaaaaatat gaacaagaga tcgacgtccg cgtacagatc gatcgcaaaa gcggtgattt
     2701 tgacactttc cgtcgctggt tagttgttga tgaagtcacc cagccgacca aggaaatcac
     2761 ccttgaagcc gcacgttatg aagatgaaag cctgaacctg ggcgattacg ttgaagatca
     2821 gattgagtct gttacctttg accgtatcac tacccagacg gcaaaacagg ttatcgtgca
     2881 gaaagtgcgt gaagccgaac gtgcgatggt ggttgatcag ttccgtgaac acgaaggtga
     2941 aatcatcacc ggcgtggtga aaaaagtaaa ccgcgacaac atctctctgg atctgggcaa
     3001 caacgctgaa gccgtgatcc tgcgcgaaga tatgctgccg cgtgaaaact tccgccctgg
     3061 cgaccgcgtt cgtggcgtgc tctattccgt tcgcccggaa gcgcgtggcg cccaactgtt
     3121 cgtcactcgt tccaagccgg aaatgctgat cgaactgttc cgtattgaag tgccagaaat
     3181 cggcgaagaa gtgattgaaa ttaaagcagc ggctcgcgat ccgggttctc gtgcgaaaat
     3241 cgcggtgaaa accaacgata aacgtatcga tccggtaggt gcttgcgtag gtatgcgtgg
     3301 cgcccgtgtt caggcggtgt ctactgaact gggtggcgag cgtatcgata tcgtcctgtg
     3361 ggatgataac ccggcgcagt tcgtgattaa cgcaatggca ccggcagacg ttgcttctat
     3421 cgtggtggat gaagataaac acacgatgga catcgccgtt gaagccggta atctggcgca
     3481 ggcgattggc cgtaacggtc agaacgtgcg tctggcttcg caactgagcg gttgggaact
     3541 caacgtgatg accgttgacg acctgcaagc taagcatcag gcggaagcgc acgcagcgat
     3601 cgacaccttc accaaatatc tcgacatcga cgaagacttc gcgactgttc tggtagaaga
     3661 aggcttctcg acgctggaag aattggccta tgtgccgatg aaagagctgt tggaaatcga
     3721 aggccttgat gagccgaccg ttgaagcact gcgcgagcgt gctaaaaatg cactggccac
     3781 cattgcacag gcccaggaag aaagcctcgg tgataacaaa ccggctgacg atctgctgaa
     3841 ccttgaaggg gtagatcgtg atttggcatt caaactggcc gcccgtggcg tttgtacgct
     3901 ggaagacctc gccgaacagg gcattgatga tctggctgat atcgaagggt tgaccgacga
     3961 aaaagccgga gcactgatta tggctgcccg taatatttgc tggttcggtg acgaagcggg
     4021 taccggcagc ggcgaaaacc tttacttcca gggccatatg ctacttcaac agcaaagagc
     4081 tccgtttccc ggacaaatgc ccaatctacc caagccaccg ctcttctggc agcaagaggc
     4141 gcagaagcag gaggcgctgc agcaggaggc gcagcagcag gaggcgcaga agaagcagca
     4201 gcaggcgcat gcgcaaatgg agcctgaacg tatcaatagc cagtagggat cctaatagtc
     4261 tagagctagc cctaggagat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     4321 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     4381 gttttttgct gaaaggagga actatatccg gatatcccgc aagaggcccg gcagtaccgg
     4441 cataaccaag cctatgccta cagcatccag ggtgacggtg ccgaggatga cgatgagcgc
     4501 attgttagat ttcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc
     4561 gcattaaagc ttatcgatga taagctgtca aacatgagaa ttacaactta tatcgtatgg
     4621 ggctgacttc aggtgctaca tttgaagaga taaattgcac tgaaatcaag aaatatttta
     4681 tctgattaat aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg
     4741 aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt tctctgagct accaactctt
     4801 tgaaccgagg taactggctt ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc
     4861 cttaaccggc gcatgacttc aagactaact cctctaaatc aattaccagt ggctgctgcc
     4921 agtggtgctt ttgcatgtct ttccgggttg gactcaagac gatagttacc ggataaggcg
     4981 cagcggtcgg actgaacggg gggttcgtgc atacagtcca gcttggagcg aactgcctac
     5041 ccggaactga gtgtcaggcg tggaatgaga caaacgcggc cataacagcg gaatgacacc
     5101 ggtaaaccga aaggcaggaa caggagagcg cacgagggag ccgccagggg gaaacgcctg
     5161 gtatctttat agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg
     5221 cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc gcggccctct cacttccctg
     5281 ttaagtatct tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc catttccgct
     5341 cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga atatatcctg
     5401 tatcacatat tctgctgacg caccggtgca gccttttttc tcctgccaca tgaagcactt
     5461 cactgacacc ctcatcagtg ccaacatagt aagccagtat acactcccct agcgctgatg
     5521 tccggcggtg cttttgccgt tacgcaccac cccgtcagta gctgaacagg agggacagct
     5581 gatagaaaca gaagccactg gagcacctca aaaacaccat catacactaa atcagtaagt
     5641 tggcagcatc acccgacgca ctttgcgccg aataaatacc tgtgacggaa gatcacttcg
     5701 cagaataaat aaatcctggt gtccctgttg ataccgggaa gccctgggcc aacttttggc
     5761 gaaaatgaga cgttgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat
     5821 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg
     5881 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
     5941 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
     6001 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
     6061 ttcttgcccg cctgatgaat gctcatccgg aattc
//
