LOCUS       sequence-289322-        6374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsTNRC6Biso1_1219-1723-del1359-1393.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6374)
  AUTHORS   Huntzinger E, Braun JE, Heimstadt S, Zekri L, Izaurralde E
  TITLE     Two PABPC1-binding sites in GW182 proteins promote miRNA-mediated 
            gene silencing.
  JOURNAL   EMBO J. 2010 Dec 15;29(24):4146-60. doi: 10.1038/emboj.2010.274
  PUBMED    21063388
REFERENCE   2  (bases 1 to 6374)
  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/emboj.2010"; journalName: "EMBO J"; date: "2010-12-15- 15";
            volume: "29"; issue: "24"; pages: "4146-60"
COMMENT     Sequence Label: pGEX-6P-1-HsTNRC6Biso1_1219-1723-del1359-1393_L
FEATURES             Location/Qualifiers
     source          1..6374
                     /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(2424..2446)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2591..2609)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2677..2781
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2782..2850,2851..3642)
                     /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(3000..3019)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3813..4401
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4302..4321
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4555..4572
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4645..4722
                     /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             4723..5805
                     /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(4742..4761)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    5818..5839
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5854..5859,5860..5877,5878..5884)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5892..5908
                     /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     5916..5932
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5916..5932
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5928..6374,1..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(5948..5965)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5948..5964)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5957..5979)
                     /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 aggcagccag
      961 gcatgaagca ctcgccctct catcctgttg ggcccaagcc gcatctggac aacatggtac
     1021 ccaacgcatt gaatgtgggg ctcccagacc ttcaaaccaa agggccaata cctggatatg
     1081 gttctggctt cagctctggc ggcatggact atggcatggt tggtgggaag gaggctggaa
     1141 ccgagtctcg ctttaaacag tggacctcca tgatggaggg gctgccctct gtagccacac
     1201 aggaagccaa tatgcacaaa aatggcgcta tagtggcccc tggtaaaacc cggggagggt
     1261 caccgtacaa ccagtttgat atcatccctg gtgacacact gggtggccat acgggtcctg
     1321 ctggtgatag ctggttacct gccaaatctc caccaacaaa taaaatcgga agtgtgctgg
     1381 ggggtacagc cacatctccc attgtagata ctgaccacca actgctgcgg gataacacca
     1441 cagggtctaa ttcttccctc aacacctcgc tgccttcacc tggtgcctgg ccctacagtg
     1501 cctctgacaa ctcctttacc aacgttcata gcacttcagc aaagttccct gattacaaat
     1561 caacatggtc cccagatccc ataggacaca accccactca tctctccaac aagatgtgga
     1621 aaaaccatat ttcctccagg aacactacac cgctgccccg cccacctcct ggtctgacca
     1681 accccaaacc atcatctccc tggagcagca cagcaccccg atcagtcagg gggtggggga
     1741 cacaggactc acggctcgcc tcggcctcta cctggagtga tggtggctca gttcgtccta
     1801 gttactggct ggttcttcac aatctcaccc cacagattga tgggtcaacc ttgagaacga
     1861 tctgcatgca gcatggccca ctgctgacat tccatctgaa tctaacccag ggcactgccc
     1921 tgatccgata cagcaccaaa caggaggcgg ccaaggccca aactgcactg cacatgtgtg
     1981 tgttgggaaa cactaccatc cttgctgagt ttgccactga tgatgaagtc agccgctttc
     2041 tggcacaagc tcagccccct acacctgcag caaccccaag tgcgccagct gcggggtggc
     2101 agtcgctgga gaccggccag aaccagtcag atcccgtggg acctgctctg aatctttttg
     2161 gtgggtccac tgggctcggg cagtggagca gcagcgctgg tggcagcagc ggggccgatc
     2221 ttgctggcgc ttcattgtgg gggcccccaa actattcttc tagcttatgg ggagtcccaa
     2281 cggtggaaga tccccatagg atgggcagcc ctgctccttt actacctggt gaccttctgg
     2341 gaggagggtc ggattcaatc taagcggccg catcgtgact gactgacgat ctgcctcgcg
     2401 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     2461 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     2521 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta taattcttga
     2581 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     2641 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2701 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2761 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     2821 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     2881 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     2941 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3001 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     3061 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3121 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3181 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3241 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3301 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     3361 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3421 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3481 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     3541 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     3601 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     3661 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3721 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3781 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3841 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3901 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     3961 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4021 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4081 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4141 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4201 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4261 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4321 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4381 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4441 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4501 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4561 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     4621 ggtatttcac accgcataaa ttccgacacc atcgaatggt gcaaaacctt tcgcggtatg
     4681 gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta
     4741 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
     4801 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
     4861 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
     4921 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
     4981 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
     5041 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
     5101 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
     5161 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt
     5221 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
     5281 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
     5341 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
     5401 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
     5461 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
     5521 gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtca
     5581 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
     5641 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
     5701 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     5761 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     5821 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     5881 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     5941 cggattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     6001 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     6061 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc
     6121 cggcaccaga agcggtgccg gaaagctggc tggagtgcga tcttcctgag gccgatactg
     6181 tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc gcccatctac accaacgtaa
     6241 cctatcccat tacggtcaat ccgccgtttg ttcccacgga gaatccgacg ggttgttact
     6301 cgctcacatt taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg
     6361 atggcgttgg aatt
//
