LOCUS       sequence-289651-        5729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsTNRC6Biso1_1379-1723del1535-1624.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5729)
  AUTHORS   Braun JE, Huntzinger E, Fauser M, Izaurralde E
  TITLE     GW182 proteins directly recruit cytoplasmic deadenylase complexes to
            miRNA targets.
  JOURNAL   Mol Cell. 2011 Oct 7;44(1):120-33.
  PUBMED    21981923
REFERENCE   2  (bases 1 to 5729)
  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.";
            date: "2011-10-7"; volume: "44(1)"; pages: "120-33"
COMMENT     Sequence Label: pGEX-6P-1-HsTNRC6Biso1_1379-1723del1535-1624_P
FEATURES             Location/Qualifiers
     source          1..5729
                     /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(1779..1801)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1946..1964)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2032..2136
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2137..2205,2206..2997)
                     /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(2355..2374)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3168..3756
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3657..3676
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3910..3927
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4000..4077
                     /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             4078..5160
                     /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(4097..4116)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    5173..5194
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5209..5214,5215..5232,5233..5239)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5247..5263
                     /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     5271..5287
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5271..5287
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5283..5729,1..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(5303..5320)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5303..5319)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5312..5334)
                     /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 atccaaaaca
      961 ttgaccctga atctgacccc tatgtcaccc caggaagtgt gctggggggt acagccacat
     1021 ctcccattgt agatactgac caccaactgc tgcgggataa caccacaggg tctaattctt
     1081 ccctcaacac ctcgctgcct tcacctggtg cctggcccta cagtgcctct gacaactcct
     1141 ttaccaacgt tcatagcact tcagcaaagt tccctgatta caaatcaaca tggtccccag
     1201 atcccatagg acacaacccc actcatctct ccaacaagat gtggaaaaac catatttcct
     1261 ccaggaacac tgcaccgctg ccccgcccac ctcctggtct gaccaacccc aaaccatcat
     1321 ctccctggag cagcacagca ccccgatcag tcagggggtg ggggacacag gactcacggc
     1381 tcgcctcggc ctctacctgg agtgatggtg gctcagttcc tgcagcaacc ccaagtgcgc
     1441 cagctgcggg gtggcagtcg ctggagaccg gccagaacca gtcagatccc gtgggacctg
     1501 ctctgaatct ttttggtggg tccactgggc tcgggcagtg gagcagcagc gctggtggca
     1561 gcagcggggc cgatcttgct ggcgcttcat tgtgggggcc cccaaactat tcttctagct
     1621 tatggggagt cccaacggtg gaagatcccc ataggatggg cagccctgct cctttactac
     1681 ctggtgacct tctgggagga gggtcggatt caatctgagc ggccgcatcg tgactgactg
     1741 acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     1801 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     1861 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     1921 gtgtataatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     1981 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     2041 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2101 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2161 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2221 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2281 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2341 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     2401 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2461 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2521 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2581 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2641 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     2701 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2761 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2821 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2881 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2941 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3001 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3061 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3121 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3181 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3241 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3301 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3361 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3421 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3481 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3541 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3601 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3661 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3721 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3781 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3841 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3901 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     3961 cttacgcatc tgtgcggtat ttcacaccgc ataaattccg acaccatcga atggtgcaaa
     4021 acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg
     4081 aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc
     4141 cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg
     4201 atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg
     4261 ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg
     4321 gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga
     4381 agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg
     4441 ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact
     4501 aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc
     4561 tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa
     4621 atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg
     4681 cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt
     4741 gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg
     4801 atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg
     4861 ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt
     4921 tatatcccgc cgtcaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg
     4981 gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc
     5041 tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg
     5101 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     5161 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     5221 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     5281 ctatgaccat gattacggat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     5341 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     5401 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     5461 gctttgcctg gtttccggca ccagaagcgg tgccggaaag ctggctggag tgcgatcttc
     5521 ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac gatgcgccca
     5581 tctacaccaa cgtaacctat cccattacgg tcaatccgcc gtttgttccc acggagaatc
     5641 cgacgggttg ttactcgctc acatttaatg ttgatgaaag ctggctacag gaaggccaga
     5701 cgcgaattat ttttgatggc gttggaatt
//
