LOCUS       sequence-289916-        6224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of CeSMG5_1-420-G91E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6224)
  AUTHORS   Jonas S, Weichenrieder O, Izaurralde E
  TITLE     An unusual arrangement of two 14-3-3-like domains in the SMG5-SMG7 
            heterodimer is required for efficient nonsense-mediated mRNA decay.
  JOURNAL   Genes Dev. 2013 Jan 15;27(2):211-25. doi: 10.1101/gad.206672.112.
  PUBMED    23348841
REFERENCE   2  (bases 1 to 6224)
  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: 
            "10.1101/gad.206672"; journalName: "Genes Dev"; date: "2013-01-15- 
            15"; volume: "27"; issue: "2"; pages: "211-25"
COMMENT     Sequence Label: pGEX-6P-1-CeSMG5_1-420-G91E_S
FEATURES             Location/Qualifiers
     source          1..6224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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)."
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2274..2296)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2441..2459)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2527..2631
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2632..2700,2701..3492)
                     /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(2850..2869)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3663..4251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4152..4171
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4405..4422
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4495..4572
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4573..5655
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4592..4611)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    5668..5689
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5704..5709,5710..5727,5728..5734)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5742..5758
                     /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)."
     primer_bind     5766..5782
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5766..5782
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5778..6224,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGITLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5798..5815)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5798..5814)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5807..5829)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc atgcaaaagt
      961 cagatgaagt cactgaaaag ttcaaacgat attgtaatca actggaaaaa tacggccaaa
     1021 ctgaaaatgt gcattctcca gtgatggcga tgttgcgcag aaaaggccga aaacagctca
     1081 ttgagattat gaaacgcgac ggggactgta caagttcaat caataaattg tggattgttg
     1141 gatattatca ccctttccag ttttttattc gagataagga gaaaaacatg gcaatagccg
     1201 tgttgctgac aatgttctgt gaagagcttc aagaaatgct ttcgcttccc gatgacaaat
     1261 acccagcatt gtggaacatg tacattggag attttcatcg ttatatgcca gacgaagaaa
     1321 ttcagaaatg cttggcagtt ggatattatt caagagcaat cgatctagat ccgaatcaag
     1381 gaagagcttt ccacgttctc gctggtctcc gagctgacct gaatgttgca cagaagctga
     1441 gactgatgat tttaggacag ctcgccgatg ctccatataa aaaaggcact gaactcttgg
     1501 aatatttgaa atttccacaa aaagaatcca ctgataaatt gatggtagat tttgtcattt
     1561 gggcattgaa cgagaaatca aaacgaatgg attatcagat gactggaatc aaaatagtaa
     1621 acgagtttaa agcagaaatc gaacagaaac tcgaattcga ttggtctctg atcatgtcaa
     1681 cgtgtcgttt agcatctaaa ttggcaatga agaaatttgg atttcaacaa ttctacaatt
     1741 gcttcgacac aatttcaacg ctgtatatta caatttattc acgtacgatt tcatccaaat
     1801 gccttctagc tgaagctatc tcatggattt ccgattcagc cgaaatactc ggacacttgg
     1861 atgagcagaa aaatgagcca catttccaga agctatcggt tttcgccaaa acgaaatgga
     1921 acgagctcaa cgacttggtt atgaatcata tcaattccgt attcacatcc atgtcactca
     1981 caatcaatcc atctatttct atgaccagct ttctgctgaa tggtccaata tccgaaccca
     2041 atgttgaatt tttgagtcaa ttaatcaatt atctggtttc cgtcgaattc ccaccaatgg
     2101 agataatcca cgatagagaa gaatcgggtc cattgctacg acgaattaat caatcggaac
     2161 aaaaacgttt ggatattcaa ataaagacac aaaatgatga agttaaccgg taggcggccg
     2221 catcgtgact gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     2281 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     2341 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     2401 cgtagcgata gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt
     2461 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     2521 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2581 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     2641 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2701 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2761 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2821 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     2881 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2941 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3001 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3061 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3121 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     3181 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3241 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3301 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3361 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3421 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3481 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3541 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3601 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3661 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3721 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3781 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     3841 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3901 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3961 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4021 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4081 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4141 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4201 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4261 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4321 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4381 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     4441 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataaa ttccgacacc
     4501 atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     4561 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     4621 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     4681 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     4741 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     4801 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     4861 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     4921 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     4981 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     5041 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     5101 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     5161 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     5221 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     5281 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     5341 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     5401 gaagacagct catgttatat cccgccgtca accaccatca aacaggattt tcgcctgctg
     5461 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     5521 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     5581 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     5641 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     5701 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     5761 tcacacagga aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc
     5821 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     5881 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     5941 tgaatggcga atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc
     6001 tggagtgcga tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg
     6061 gttacgatgc gcccatctac accaacgtaa cctatcccat tacggtcaat ccgccgtttg
     6121 ttcccacgga gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc
     6181 tacaggaagg ccagacgcga attatttttg atggcgttgg aatt
//
