LOCUS       sequence-289391-        8403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6C_1-1207.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8403)
  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 8403)
  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-HsTNRC6C_1-1207_M
FEATURES             Location/Qualifiers
     source          1..8403
                     /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     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     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"
     CDS             3679..3696
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5191..5312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5228..5247)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5282..5301
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5319..5774)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5456..5477)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5668..5687
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5801..5905
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5907..6264
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5907..6098
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5930..5950)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6115..6250
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6299..7093
                     /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             6299..7093
                     /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(6353..6372)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6963..6982
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7281..7300)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7325..7372
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7701..8289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8190..8209
                     /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 gagctacagg gagtgcccag ggcaacttca ctggacatac
     1441 caagaagaca aatggcaata atggcaccaa tggcgcactc gtccaaagcc cttctaatca
     1501 gagtgccctt ggagcagggg gagcgaacag taatggaagt gcggccagag tgtggggtgt
     1561 agccacaggc tccagctctg gcctggctca ctgctctgtc agtggtgggg atggaaaaat
     1621 ggacactatg attggagatg ggagaagtca gaattgctgg ggtgcttcca actccaatgc
     1681 tggcattaat cttaacctta atcctaatgc caacccagct gcctggcctg tacttggaca
     1741 tgaaggaacc gtggcgacag gcaacccttc cagtatttgc agtccagtca gtgccatagg
     1801 tcaaaatatg ggcaaccaga acgggaaccc aacaggcact ttaggtgctt ggggaaactt
     1861 gctgccacaa gagagcacag aaccacaaac gtccacttct cagaatgtgt ctttcagcgc
     1921 acaacctcag aaccttaaca ctgatggacc aaataacact aaccccatga actcttcacc
     1981 caaccctatc aatgcaatgc agacaaatgg actgccaaac tggggcatgg ctgttggtat
     2041 gggggccatc atcccgcccc acctgcaagg ccttcctggt gctaatggat catcagtttc
     2101 tcaagtcagt gggggcagtg ctgaaggaat aagcaattct gtgtggggac tgtccccagg
     2161 taaccctgcc acaggaaata gcaattctgg gttcagtcag gggaatggag acactgtgaa
     2221 ctcagcatta agtgctaaac aaaatggatc cagcagtgct gtgcaaaagg aaggaagtgg
     2281 aggaaatgct tgggattcag gacctcctgc tggtcctgga atactcgcct ggggaagggg
     2341 cagtggcaac aatggcgttg gtaatatcca ttcaggagct tggggccacc ccagccgaag
     2401 cacctctaac ggtgtgaatg gggaatgggg aaagccccca aaccagcatt ccaacagtga
     2461 catcaatggg aaaggatcaa cagggtggga gagtcctagt gtcaccagcc agaaccctac
     2521 cgtacagcct ggtggtgaac acatgaactc ctgggccaaa gcggcatctt ctggaactac
     2581 agcaagtgaa ggaagtagtg atggttctgg caaccacaat gaaggaagca ctgggaggga
     2641 aggaacggga gaaggccgaa ggcgagataa agggattata gaccaagggc acatccagtt
     2701 gccaaggaat gatcttgacc caagagttct gtctaatact ggttggggac agactcctgt
     2761 aaagcaaaac actgcctggg aatttgaaga atcccctagg tctgaaagga aaaatgacaa
     2821 tgggacagag gcctggggtt gtgcagctac tcaggcttca aactcagggg ggaagaacga
     2881 tgggtccatc atgaacagta caaatacctc ttcagtatct gggtgggtca acgcgccacc
     2941 tgccgctgtg ccagcaaaca caggttgggg agacagcaac aacaaagcgc caagtggccc
     3001 gggggtttgg ggggactcga taagctctac tgctgttagt actgctgctg ctgccaagag
     3061 tggccatgct tggagtgggg ccgcaaatca ggaggacaag tcacccacct ggggtgagcc
     3121 tccaaagccc aaatcccaac actggggaga tggacaaaga tcaaatccag cctggagtgc
     3181 aggaggggga gattgggcag attcatcgtc tgtccttgga cacttggggg atgggaaaaa
     3241 aaatggatct ggatgggatg ctgacagtaa taggtcaggg tctggttgga atgacaccac
     3301 gagatctggg aacagtggct ggggcaacag cacaaataca aaggccaatc caggtacaaa
     3361 ctggggggag actttaaaac ctggccccca acagaactgg gctagcaaac cccaagacaa
     3421 caatgtgagt aactggggag gagctgcttc tgtgaaacag acaggaacag ggtggatcgg
     3481 ggggccggta ccggtcaaac agaaggacag cagtgaagca actggctggg aagaaccctc
     3541 tccaccgtcc attcgccgca aaatggaaat tgatgatggt acctcagctt ggggggaccc
     3601 aagcaactat aacaataaaa ctgtaaacat gtgggataga aacaacccgg tcatccagag
     3661 cagtaccacg accaatacca ccaccaccac caccactacc acgagcaaca ccacacacag
     3721 ggtcgagacg ccgcccccgc accaggctgg tactcagctg aatcgatcac cgttgcttgg
     3781 tccaggtagg aaagtttcat caggctgggg agaaatgcct aatgttcact caaagactga
     3841 aaactcttgg ggagaaccat cctccccttc taccctggtg gataatggca cagcagcatg
     3901 ggggaagcca cccagcagtg gcagcgggtg gggagatcac cctgccgagc cgccggtggc
     3961 atttggaaga gctggcgcac ctgttgctgc ctcagccctg tgcaaaccag cttcaaaatc
     4021 tatgcaagaa ggctggggca gtggtgggga tgaaatgaac ctcagtacca gccagtggga
     4081 ggatgaagaa ggggacgtgt ggaataatgc tgcttcccaa gaaagcacct cctcctgcag
     4141 ctcctggggg aacgccccca aaaaaggact tcaaaagggc atgaagacgt ctggcaagca
     4201 ggatgaggcc tggatcatga gccggctgat caaacaactc acagacatgg gcttcccgag
     4261 agagccagct gaggaggcct tgaagagtaa caatatgaat cttgatcagg ccatgagcgc
     4321 tctgctggaa aagaaggtgg acgtggacaa gcgtgggctg ggagtgaccg accataatgg
     4381 aatggccgcc aagcccctcg gctgccgccc gccaatctcc aaagagtctt ccgtggaccg
     4441 ccccaccttt cttgacaagg atggcggcct cgtggaagag cccacgcctt caccgttctt
     4501 gccttcccca agcctgaagc tccccctttc acacagtgca ctccccagtc aggccctggg
     4561 tgggattgcc tccgggctgg gcatgcaaaa cttgaattct tctagacaga taccgagtgg
     4621 caatctgggt atgtttggca atagtggagc agcacaagcc aggaccatgc agcagccgcc
     4681 acagccacca gtgcagcctc ttaactcttc ccagcccagt ctccgtgctc aagtgcctca
     4741 gtttctatcc cctcaggttc aagcacagct tttgcagttt gcagcaaaaa acattggtct
     4801 caaccctgca ctattaacct cgccaattaa tcctcaacat atgacgatgt tgaaccagct
     4861 ctatcagctg cagctggcat accaacgttt acaaatccag cagcagatgt tacaggccca
     4921 gcgtaatgtg tccggatcca tgagacaaca ggagcagcaa gttgcgcgca caatcactaa
     4981 tctgcagcag cagatccagc agcaccagcg ccagctggcc tagaagcttc gaattctgca
     5041 gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat
     5101 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     5161 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     5221 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5281 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     5341 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     5401 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     5461 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5521 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     5581 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5641 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5701 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5761 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5821 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5881 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     5941 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     6001 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     6061 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     6121 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     6181 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     6241 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     6301 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     6361 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     6421 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     6481 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     6541 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     6601 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     6661 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     6721 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     6781 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     6841 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     6901 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     6961 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     7021 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     7081 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     7141 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     7201 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     7261 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     7321 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     7381 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     7441 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     7501 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     7561 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7621 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7681 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7741 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7801 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     7861 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7921 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7981 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     8041 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     8101 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     8161 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     8221 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     8281 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     8341 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     8401 cat
//
