LOCUS       sequence-289334-        8410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6Biso1_1-1218.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8410)
  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 8410)
  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: pT7-EGFP-C1-HsTNRC6Biso1 1-1218_L
FEATURES             Location/Qualifiers
     source          1..8410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(710..731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(971..990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1318..1339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5198..5319
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5235..5254)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5289..5308
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5326..5781)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5463..5484)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5675..5694
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5808..5912
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5914..6271
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5914..6105
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5937..5957)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6122..6257
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6306..7100
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6306..7100
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6360..6379)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6970..6989
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7288..7307)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7332..7379
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7708..8296
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8197..8216
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      661 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      721 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      781 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      841 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      961 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1021 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1081 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1141 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1261 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1381 gtccggactc agatctcgag ctcaagcttc gatgagagag aaggagcaag aaagggaaga
     1441 acagttaatg gaagacaaga aaaggaagaa agaggataaa aagaaaaaag aagccactca
     1501 gaaggtcacg gaacaaaaaa ccaaagtgcc cgaagtgacg aaaccaagtt taagccaacc
     1561 aacggccgcc agcccaattg gcagctctcc atcgccacca gtcaatggtg gcaacaatgc
     1621 caaaagggtg gcagtgccga acggacaacc gccaagcgcc gcccgctaca tgcctcggga
     1681 ggtgccgccg cgattccgtt gccagcagga ccacaaagtg ttactaaaac gtgggcagcc
     1741 ccctccaccg tcctgcatgc tccttggggg tggggcaggg cctcctccct gcacagcacc
     1801 tggagcaaac ccaaacaacg cacaagtgac aggagcgctg ctgcagagtg agagtgggac
     1861 tgcgccagac tcaacccttg gaggtgctgc tgcttcaaat tatgcaaatt ccacttgggg
     1921 ctcgggagcc tcctccaaca acggcacctc ccccaaccca attcacatct gggacaaggt
     1981 gattgtagac gggtctgaca tggaagagtg gccttgtatt gccagcaaag acactgaatc
     2041 ttcttccgaa aacaccaccg ataacaacag tgcctcgaac cctggctctg agaagagcac
     2101 tctgccagga agcaccacta gtaacaaagg aaaagggagc cagtgccagt ctgcaagttc
     2161 tgggaacgaa tgtaatcttg gggtctggaa atctgaccct aaggctaaat ctgttcaatc
     2221 ttccaactct actacagaga acaacaatgg actaggaaat tggaggaatg tgagtggtca
     2281 ggatagaatt ggacctggct ctggcttcag caactttaac ccaaatagca acccatctgc
     2341 ctggccagca ctggtccaag aaggaacttc taggaaaggg gcattggaaa cagataatag
     2401 taattccagt gcacaggtta gcacagtagg tcagacatcc agggaacagc agtcaaagat
     2461 ggaaaatgcg ggtgttaatt ttgttgtctc tggcagagaa caggctcaaa ttcataacac
     2521 tgatggacca aaaaatggaa acactaactc cttgaactta agttcaccaa accccatgga
     2581 gaataaggga atgccctttg gaatgggctt ggggaacacc tccaggagca ctgatgcccc
     2641 ttcacaaagc actggagatc gaaagactgg gagtgttgga tcttggggtg cagctagggg
     2701 gccttctgga actgacacag tctctggaca aggcaattct ggaaacaatg ggaacaatgg
     2761 aaaagagaga gaggactcct ggaaaggagc ttctgttcag aaatcaactg ggtcaaaaaa
     2821 tgactcttgg gacaacaata acaggtctac gggtgggtcc tggaactttg gcccccagga
     2881 ctctaatgac aacaaatggg gtgaagggaa caaaatgaca tctggggtct ctcagggaga
     2941 atggaaacag ccgactgggt ctgatgagtt gaaaattgga gaatggagtg gtccaaacca
     3001 accaaattct agcactggag catgggacaa tcaaaagggc caccccctcc ctgaaaacca
     3061 aggcaatgcc caggctccct gttggggaag atcttccagc tccacaggaa gtgaagttgg
     3121 aggtcaaagc actggaagca accacaaagc aggaagtagt gacagtcata actctggccg
     3181 tcggtcgtac aggcccacac atcctgattg tcaggctgtc ttgcagactc ttttgagccg
     3241 aactgatttg gaccccaggg tgctctcaaa cactggctgg ggccaaactc aaattaagca
     3301 ggacacagtg tgggacattg aagaggtgcc aaggcctgag gggaaatctg acaaaggaac
     3361 tgaggggtgg gagagcgctg ccacacagac caagaactca gggggctggg gagatgcacc
     3421 cagccaaagc aatcaaatga agtctggatg gggggagctc tcagcctcta cagagtggaa
     3481 agaccccaag aacacaggag gctggaatga ctacaagaac aacaactctt ccaactgggg
     3541 aggaggacga cctgatgaaa agacaccttc ctcttggaat gagaatccca gcaaggatca
     3601 ggggtgggga ggtggacgcc agcccaatca aggatggtct tctggaaaga atggttgggg
     3661 ggaggaagtc gatcagacaa aaaacagcaa ttgggaaagt tctgcaagta aacctgtgtc
     3721 tgggtggggt gaaggagggc agaatgaaat cgggacttgg ggtaatggtg gcaatgcaag
     3781 cctagcttca aaaggtgggt gggaggattg caaaagatcc ccagcatgga atgagacggg
     3841 ccgacagccc aattcctgga ataaacaaca ccaacagcag cagcccccac agcagccgcc
     3901 gccaccacaa ccagaggctt ctggttcgtg gggaggccca cccccaccac ctccaggcaa
     3961 cgttcgacct tccaattcca gctggagcag cgggccacag cctgcaacac ctaaggatga
     4021 ggaacccagt ggttgggaag agccatcccc acagtcaatt agtcggaaaa tggacattga
     4081 tgatggcact tcagcatggg gagaccctaa cagttataac tacaagaatg tgaatctgtg
     4141 ggataagaat tcccaagggg gcccagcacc tcgagaacca aacctgccca ccccaatgac
     4201 cagtaaatcg gcatcagatt ccaaatctat gcaagacggc tggggggaga gtgacgggcc
     4261 agtcacagga gctcgccatc ccagctggga agaggaggag gatggaggag tctggaacac
     4321 cactggctct cagggcagtg cttcctccca caactcagca agctggggac aaggaggaaa
     4381 gaaacaaatg aagtgctcac tcaaaggagg aaacaatgat tcatggatga atcctcttgc
     4441 caaacagttt tcaaatatgg gattgctgag tcagactgaa gataatccaa gcagcaaaat
     4501 ggatttgtct gtaggaagcc tttcagataa aaaatttgat gtggacaagc gagcgatgaa
     4561 tctcggggat tttaatgata tcatgaggaa ggatcgatct gggttccgtc cacctaattc
     4621 caaagacatg ggaaccacag atagtgggcc ttattttgag aagggcggta gtcgtggttt
     4681 gtttggaaac agcacagcac aatcgagagg tctgcacaca cccgtgcggc cactaaattc
     4741 ttctcccagt ctccgggcgc aagtgcctcc ccagtttatt tccccccagg tttctgcctc
     4801 aatgctcaag cagtttccca acagtggcct gagtccaggt cttttcaatg tggggcccca
     4861 gttatctcct caacaaattg ccatgctgag ccagcttcca caaattcccc agtttcagtt
     4921 ggcatgtcag cttctcttgc agcagcagca acagcagcgg ttgttacaga accagagaaa
     4981 gatttctcaa gctgtacgcc aacagcaaga gcagcagctg gctcgaatgg tgagtgcact
     5041 gcagcagcag cagcagcagc agaggtgagg atccaccgga tctagataac tgatcataat
     5101 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     5161 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     5221 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5281 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta
     5341 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     5401 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     5461 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     5521 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     5581 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     5641 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     5701 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     5761 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5821 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5881 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     5941 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     6001 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     6061 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     6121 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     6181 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     6241 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     6301 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     6361 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     6421 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     6481 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     6541 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     6601 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     6661 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     6721 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     6781 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     6841 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     6901 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     6961 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     7021 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     7081 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     7141 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     7201 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     7261 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     7321 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     7381 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     7441 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     7501 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     7561 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     7621 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     7681 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     7741 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     7801 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     7861 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     7921 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     7981 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     8041 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     8101 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     8161 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     8221 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     8281 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     8341 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     8401 cgccatgcat
//
