LOCUS       sequence-289012-        6523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsSmg6_1-515.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6523)
  AUTHORS   Kashima I, Jonas S, Jayachandran U, Buchwald G, Conti E, Lupas AN, 
            Izaurralde E
  TITLE     SMG6 interacts with the exon junction complex via two conserved 
            EJC-binding motifs (EBMs) required for nonsense-mediated mRNA decay.
  JOURNAL   Genes Dev. 2010 Nov 1. 24(21):2440-50.
  PUBMED    20930030
REFERENCE   2  (bases 1 to 6523)
  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"; journalName: "Genes Dev. 
            2010 Nov 1. 24(21):2440-50."
COMMENT     Sequence Label: pGEX-6P-1-HsSmg6_1-515_I
FEATURES             Location/Qualifiers
     source          1..6523
                     /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(2573..2595)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2740..2758)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2826..2930
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2931..2999,3000..3791)
                     /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(3149..3168)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3962..4550
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4704..4721
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4794..4871
                     /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             4872..5954
                     /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(4891..4910)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    5967..5988
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6003..6008,6009..6026,6027..6033)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6041..6057
                     /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     6065..6081
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6065..6081
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6077..6523,1..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(6097..6114)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6097..6113)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6106..6128)
                     /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 ccggaattga
      961 tggcggaagg gctggagcgt gtgcggatct ccgcgtcgga gctgcgcggg atcctggcta
     1021 ctctggcccc gcaggccggg agcagagaaa acatgaagga attaaaggag gccaggccgc
     1081 gcaaagataa caggcgtcca gatctggaaa tctataagcc tggcctttct cggctaagga
     1141 acaagcccaa aatcaaggaa ccccctggga gtgaggaatt caaagatgaa attgttaatg
     1201 accgagattg ctctgctgtt gaaaatggta cacagcccgt taaagatgtc tgcaaggaac
     1261 tgaacaacca agagcagaat ggtcctatag acccagaaaa taatcgggga caagaatcct
     1321 ttcctaggac tgctggacaa gaggatcgta gtctaaaaat tatcaaaaga acaaagaaac
     1381 ccgacctgca gatctatcag cctggacgac gtttgcagac tgttagcaaa gaatccgcca
     1441 gtcgggtgga ggaggaagaa gtcctcaacc aggtagaaca actgagagta gaggaagatg
     1501 agtgtagggg aaatgttgcg aaggaggaag ttgcgaataa accagacagg gccgagatag
     1561 aaaagagccc aggtggtggg agagtagggg ctgcaaaagg agaaaaagga aagaggatgg
     1621 gaaaagggga gggggtgagg gaaacccacg acgacccggc ccgcgggagg ccgggctccg
     1681 caaagcgcta ctcccgctca gacaaacgaa ggaatcgcta ccgcacgcgc agcaccagct
     1741 cagctggcag caacaacagc gctgagggag ctggcctgac ggataatgga tgtcgccgcc
     1801 gccgacagga taggaccaag gagaggccac cactgaagaa gcaagtgtct gtgtcctcaa
     1861 ccgattcctt agacgaggac agaattgatg agcctgatgg attaggaccc aggagaagtt
     1921 cagaaaggaa gagacattta gaaagaaact ggtctggccg tggggagggt gagcagaaaa
     1981 ccagtgctaa agaatatcga ggcactcttc gtgtcacttt cgatgcagaa gccatgaaca
     2041 aagagtctcc catggtgagg tcagccaggg atgatatgga tagaggaaag cctgacaaag
     2101 gcttgagcag tgggggcaaa ggctctgaga agcaggagtc caaaaacccg aaacaagaac
     2161 ttcggggtcg tggtcgtggc attctgattt tgcctgccca taccacccta tctgtcaatt
     2221 cagcaggttc tccagagtcc gcgcctttgg gacctcggct tttgtttgga tctggtagta
     2281 agggatctcg gagttggggc cgtggaggca ccacacgccg attgtgggac ccaaacaatc
     2341 ctgatcagaa acctgctcta aagactcaga cgccccagct acatttcttg gacactgatg
     2401 atgaagtcag ccctacatct tggggtgact cacgccaggc tcaggcatct tactataagt
     2461 ttcaaaactc tgacaacccc tattattacc cccggacacc aggctgactc gagcggccgc
     2521 atcgtgactg actgacgatc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     2581 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     2641 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     2701 gtagcgatag cggagtgtat aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
     2761 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     2821 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2881 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2941 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3001 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3061 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3121 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     3181 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3241 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3301 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3361 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3421 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     3481 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3541 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3601 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3661 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3721 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3781 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3841 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3901 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3961 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4021 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4081 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     4141 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4201 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4261 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4321 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4381 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4441 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4501 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4561 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4621 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4681 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     4741 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataaat tccgacacca
     4801 tcgaatggtg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     4861 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     4921 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     4981 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     5041 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     5101 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     5161 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     5221 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     5281 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     5341 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
     5401 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     5461 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
     5521 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
     5581 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
     5641 ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg
     5701 aagacagctc atgttatatc ccgccgtcaa ccaccatcaa acaggatttt cgcctgctgg
     5761 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     5821 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
     5881 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     5941 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     6001 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     6061 cacacaggaa acagctatga ccatgattac ggattcactg gccgtcgttt tacaacgtcg
     6121 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     6181 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     6241 gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa gcggtgccgg aaagctggct
     6301 ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg
     6361 ttacgatgcg cccatctaca ccaacgtaac ctatcccatt acggtcaatc cgccgtttgt
     6421 tcccacggag aatccgacgg gttgttactc gctcacattt aatgttgatg aaagctggct
     6481 acaggaaggc cagacgcgaa ttatttttga tggcgttgga att
//
