LOCUS       sequence-288716-        6743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmGW182_1-593.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6743)
  AUTHORS   Behm-Ansmant I, Rehwinkel J, Doerks T, Stark A, Bork P, Izaurralde E
  TITLE     mRNA degradation by miRNAs and GW182 requires both CCR4:NOT 
            deadenylase and DCP1:DCP2 decapping complexes.
  JOURNAL   Genes Dev. 2006 Jul 15. 20(14):1885-98.
  PUBMED    16815998
REFERENCE   2  (bases 1 to 6743)
  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. 
            2006 Jul 15. 20(14):1885-98."
COMMENT     Sequence Label: pGEX-5X-3-GST-DmGW182_1-593_F
FEATURES             Location/Qualifiers
     source          1..6743
                     /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             921..932
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2793..2815)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2960..2978)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3046..3150
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3151..3219,3220..4011)
                     /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(3369..3388)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4182..4770
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4671..4690
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4924..4941
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5014..5091
                     /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             5092..6174
                     /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
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5111..5130)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    6187..6208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6223..6228,6229..6246,6247..6253)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6261..6277
                     /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     6285..6301
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6285..6301
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6297..6743,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6317..6334)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6317..6333)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6326..6348)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttatcga 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 atcgaaggtc gtgggatccc caggaattct atgcgtgaag
      961 cccttttttc ccaagatggc tggggctgtc agcatgttaa ccaggatact aattgggaag
     1021 ttcccagttc gccagaacca gccaataagg atgcacccgg tccaccaatg tggaagccaa
     1081 gcattaacaa tggtactgat ctttgggagt ccaatttgag aaacggaggt cagccggccg
     1141 cacagcaagt tccaaagccg tcgtggggtc atacaccatc ctctaactta ggtggaacat
     1201 ggggtgagga cgacgatggc gccgatagta gtagtgtgtg gactggagga gctgttagca
     1261 acgcgggatc cggagctgca gtgggagtaa accaagccgg agttaatgtc ggtccaggcg
     1321 gtgttgtttc gtctggcgga cctcagtggg gacaaggtgt cgttggcgtc ggacttggat
     1381 caactggagg taacgggtca agcaatataa ctggatcgtc tggagtcgca acaggtagta
     1441 gcggaaactc cagcaacgct ggtaacggtt ggggagaccc tcgtgaaata cgccctttgg
     1501 gagttggtgg ctccatggat attcgaaatg ttgaacatcg cggcggtaac ggttctggag
     1561 caacttcgag cgatccacga gacattcgca tgatcgatcc gcgtgaccct attcgaggag
     1621 atccccgtgg aatatctggt cgtcttaatg ggacctctga aatgtggggt catcatccac
     1681 aaatgtccca taaccagttg caaggtatca acaaaatggt tggtcaaagt gtagcaactg
     1741 ccagcaccag tgtcggaaca tctggctcgg gcatcggtcc tggaggtccc ggtcctagta
     1801 cagtatcagg caatatccca acacagtggg ggcctgctca accggtaagc gttggtgtaa
     1861 gtggtcccaa agacatgtca aaacagataa gtggatggga ggaaccatca ccaccgcctc
     1921 agcgtcgcag tattcctaac tacgatgatg gtacatcgtt gtggggtcag caaactcgtg
     1981 ttcccgctgc aagcggtcac tggaaagaca tgactgattc gataggtcgt agtagtcatc
     2041 tcatgcgtgg ccaaagccaa acgggaggta taggaatagc cggcgttgga aatagcaatg
     2101 ttccagtggg agccaatcca agtaatccta taagcagtgt agttggacct caagcccgga
     2161 ttccatctgt gggcggcgta caacacaaac cagacggcgg cgctatgtgg gtgcattccg
     2221 gcaatgtagg tggcagaaat aatgttgctg ctgttactac ttggggagat gacactcata
     2281 gcgttaatgt cggcgctccc agcagtggca gtgtatccag caacaattgg gttgatgaca
     2341 agtccaactc aaccttggca caaaactctt ggagcgaccc ggcccctgtt ggagttagtt
     2401 ggggcaataa gcaaagcaaa ccgccaagca atagtgcttc atcaggttgg agcactgctg
     2461 cgggcgtggt ggatggggtt gatctaggat ctgagtggaa cacgcacggg gggattattg
     2521 gaaaatctca gcaacaacaa aaactagcgg gacttaacgt gggaatggtg aacgtaatta
     2581 acgcggagat cattaagcaa agcaagcaat acaggatcct tgtcgagaac ggctttaaaa
     2641 aggaagatgt agagcgggca ttagtgattg ctaatatgaa catcgaagag gcagccgata
     2701 tgctccgtgc caactcatcc caatcaatgt aggcggccgc atcgtgactg actgacgatc
     2761 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     2821 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     2881 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     2941 aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3001 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3061 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3121 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3181 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3241 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3301 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3361 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     3421 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3481 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3541 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3601 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3661 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     3721 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3781 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3841 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3901 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3961 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4021 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4081 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4141 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4201 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4261 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4321 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4381 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4441 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4501 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4561 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4621 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4681 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4741 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4801 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4861 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4921 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     4981 catctgtgcg gtatttcaca ccgcataaat tccgacacca tcgaatggtg caaaaccttt
     5041 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
     5101 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
     5161 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
     5221 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
     5281 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
     5341 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
     5401 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
     5461 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
     5521 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
     5581 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
     5641 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
     5701 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
     5761 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
     5821 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
     5881 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
     5941 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
     6001 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
     6061 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     6121 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     6181 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     6241 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     6301 ccatgattac ggattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     6361 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     6421 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg
     6481 cctggtttcc ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg
     6541 ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca
     6601 ccaacgtaac ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg
     6661 gttgttactc gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa
     6721 ttatttttga tggcgttgga att
//
