LOCUS       sequence-437536-        6385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsRck.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6385)
  AUTHORS   Tritschler F, Braun JE, Eulalio A, Truffault V, Izaurralde E, 
            Weichenrieder O
  TITLE     Structural basis for the mutually exclusive anchoring of P body 
            components EDC3 and Tral to the DEAD box protein DDX6/Me31B.
  JOURNAL   Mol Cell. 2009 Mar 13. 33(5):661-8.
  PUBMED    19285948
REFERENCE   2  (bases 1 to 6385)
  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: "Mol Cell. 
            2009 Mar 13. 33(5):661-8."
COMMENT     Sequence Label: pGEX-6P-1-HsRCK_E
FEATURES             Location/Qualifiers
     source          1..6385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(213..218,219..234,235..241)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    249..265
                     /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             288..941
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     899..921
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             948..971
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2465..2487)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2632..2650)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2718..2822
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2823..2891,2892..3683)
                     /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(3041..3060)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3854..4442
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4343..4362
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4596..4613
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4686..4763
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4764..5846
                     /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(4783..4802)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5895..5900,5901..5918,5919..5925)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5933..5949
                     /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     5957..5973
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5957..5973
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5969..6385,1..111)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5989..6006)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5989..6005)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5998..6020)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 acgcgaatta tttttgatgg cgttggaatt agcttatcga ctgcacggtg caccaatgct
       61 tctggcgtca ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta aatcactgca
      121 taattcgtgt cgctcaaggc gcactcccgt tctggataat gttttttgcg ccgacatcat
      181 aacggttctg gcaaatattc tgaaatgagc tgttgacaat taatcatcgg ctcgtataat
      241 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agtattcatg tcccctatac
      301 taggttattg gaaaattaag ggccttgtgc aacccactcg acttcttttg gaatatcttg
      361 aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa
      421 agtttgaatt gggtttggag tttcccaatc ttccttatta tattgatggt gatgttaaat
      481 taacacagtc tatggccatc atacgttata tagctgacaa gcacaacatg ttgggtggtt
      541 gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc ggttttggat attagatacg
      601 gtgtttcgag aattgcatat agtaaagact ttgaaactct caaagttgat tttcttagca
      661 agctacctga aatgctgaaa atgttcgaag atcgtttatg tcataaaaca tatttaaatg
      721 gtgatcatgt aacccatcct gacttcatgt tgtatgacgc tcttgatgtt gttttataca
      781 tggacccaat gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag
      841 ctatcccaca aattgataag tacttgaaat ccagcaagta tatagcatgg cctttgcagg
      901 gctggcaagc cacgtttggt ggtggcgacc atcctccaaa atcggatctg gaagttctgt
      961 tccaggggcc cctgggatcc atgggtctgt ccagtcaaaa tggtcagctg agaggccctg
     1021 tgaaacccac tggtggccct ggaggagggg gcacacagac acagcaacag atgaaccagc
     1081 tgaaaaacac caacacaatc aataatggca ctcagcagca agcacagagt atgaccacca
     1141 ctattaaacc tggtgatgac tggaaaaaga ctttaaaact ccctccaaag gatctaagaa
     1201 tcaaaacttc ggatgtgacc tccacaaaag gaaatgagtt tgaagattac tgtttgaaac
     1261 gggagttact gatgggaatt tttgaaatgg gctgggaaaa gccatctcct attcaggagg
     1321 agagcattcc cattgcttta tctggtaggg atatcttagc tagagcaaaa aatggaacag
     1381 gcaagagcgg tgcctacctc attcccttac ttgaacggct agacctgaag aaggacaata
     1441 tacaagcaat ggtgattgtt cccactagag aacttgctct acaggtcagt caaatttgca
     1501 tccaggtcag caaacacatg ggaggggcca aagtgatggc aaccacagga ggaaccaatt
     1561 tacgagatga cataatgagg cttgatgata cagtgcacgt ggtgattgct acccctggga
     1621 gaatcctgga tcttattaag aaaggagtag caaaggttga tcatgtccag atgatagtat
     1681 tggatgaggc agataagttg ctgtcacagg attttgtgca gataatggag gatattattc
     1741 tcacgctacc taaaaacagg cagattttac tatattccgc tactttccct cttagtgtac
     1801 agaagttcat gaattcccat ttgcagaaac cctatgagat taacctgatg gaggaactaa
     1861 ctctgaaggg agtaacccag tactacgcat atgtaactga gcgccaaaaa gtacactgcc
     1921 tcaacacact tttctccagg cttcagataa accagtcgat cattttctgt aactcctctc
     1981 agcgagttga attgctagcc aagaagattt ctcaactggg ttattcttgc ttctatattc
     2041 atgctaaaat gaggcaggaa catcgaaatc gtgtatttca tgatttccga aatggcttat
     2101 gccgcaatct tgtttgcact gatctgttta cccgaggtat tgatatacaa gctgtgaatg
     2161 tggtaataaa ctttgatttc ccaaagctgg cagagaccta tctccatcgt attggaagat
     2221 caggtcgctt tggtcatctt ggcttagcca tcaacttgat cacatatgat gatcgcttca
     2281 acctgaaaag tattgaggag cagctgggaa cagaaattaa acctattccg agcaacattg
     2341 ataagagcct gtatgtggca gaataccaca gcgagcctgt agaagatgag aaaccttaac
     2401 tcgagcggcc gcatcgtgac tgactgacga tctgcctcgc gcgtttcggt gatgacggtg
     2461 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     2521 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
     2581 tgacccagtc acgtagcgat agcggagtgt ataattcttg aagacgaaag ggcctcgtga
     2641 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     2701 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2761 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2821 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2881 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2941 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3001 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3061 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3121 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3181 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3241 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3301 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3361 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3421 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3481 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3541 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3601 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3661 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3721 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3781 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3841 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3901 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3961 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     4021 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4081 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4141 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4201 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4261 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4321 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4381 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4441 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4501 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     4561 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     4621 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcataa
     4681 attccgacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
     4741 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
     4801 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
     4861 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
     4921 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
     4981 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
     5041 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
     5101 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
     5161 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
     5221 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
     5281 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
     5341 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
     5401 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
     5461 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
     5521 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
     5581 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
     5641 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     5701 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacg ctagcgccca
     5761 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     5821 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat
     5881 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     5941 ggataacaat ttcacacagg aaacagctat gaccatgatt acggattcac tggccgtcgt
     6001 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     6061 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     6121 gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt ccggcaccag aagcggtgcc
     6181 ggaaagctgg ctggagtgcg atcttcctga ggccgatact gtcgtcgtcc cctcaaactg
     6241 gcagatgcac ggttacgatg cgcccatcta caccaacgtg acctatccca ttacggtcaa
     6301 tccgccgttt gttcccacgg agaatccgac gggttgttac tcgctcacat ttaatgttga
     6361 tgaaagctgg ctacaggaag gccag
//
