LOCUS       sequence-478954-        8397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6A_1-1208.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8397)
  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 8397)
  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-HsTNRC6A_1-1208_M
FEATURES             Location/Qualifiers
     source          1..8397
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(1960..2670,2671..2673,2674..2676))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1960..2670,2671..2673,2674..2676))
                     /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(2002..2023)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2351..2370
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2610..2631
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2673..2682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2709..2728)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2710..2728)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2773..2976)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2802..2822)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2977..3280
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3455..4043)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3535..3554)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4372..4419
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4444..4463
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4651..5445)
                     /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             complement(4651..5445)
                     /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(4762..4781)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5372..5391
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5480..5837)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5494..5629
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5646..5837
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5794..5814
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5839..5943)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5970..6425
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6057..6076)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6267..6288
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6432..6553
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6443..6462)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6497..6516
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 tccccaagtg aaagaactag ttttaccaag ttcgttccaa ccggaaacgg acctgtcact
       61 gtcactacct cctgagctgg atttcttatt ggcctcaccc caatggttat tccttgaagt
      121 ttctccccag ttatcactgg cagaaacaga ccacccttgg cttgattttt gtccatcacc
      181 ccatccctgt ttattctttt gcgagtcatt ccatgcagcc ttctcctctt tgcaattccc
      241 ccactgattg ctcttgacca ttcctgtagc agcagaatca tcttcccacc ccccctggca
      301 gtttgagcct ttggaatctc cccacctcag agcaggtttg ggatcgcccc accctgatac
      361 agatgaggta tcattaccat tagggctagt cttatctata catgcccctg agttaaaagt
      421 ctgtgttgca gagcttcccc aggcctctgt cccattgtca gtctttcgtt cccctctagg
      481 tgatgtttct gtatcccagg cagtattctg cttaatagga gtctgtcccc aaccagagtt
      541 ggacaggaca cgtggatcta agtcagttct gtttacaatg ctttggagta atgtgtgctg
      601 atcaattttt cttctgtctc tactctgact ttccccagtt cctttttcct caaggcgtcc
      661 agtgctttct gtactaccat cgctctccac tgtacctcct gttttggccc acacactgct
      721 ttgctcattt gtctgagatg tggcagaacc ctgatcctct tcctcagtag atttccatcc
      781 atttgtaaac gtcttaccat tgccatttgc actatcattg gaatgctgat tgctaggcag
      841 tttgttccac tcaacgctgg gtaaattagt gccagtgttt tgtgcagggg ttccccatcc
      901 tcttcgactt cctccagaat ttgtgccatt tccacttccc catgatgtac tctgggagtt
      961 tgctgcacca gattcccaca caccacctcc tttgtttgtg ttaacttgaa agttagtgcc
     1021 cataggacca tttacgccag gctgcattag agttgcattc acagtgtcac cattagcttg
     1081 gccattaggg cctgaacatt tgtctccaga gtaattagaa ccataggcac cccatgtagt
     1141 accataagag cctccacttt ttgactctcc attgctaagg tgagaaaggg aagtgccatt
     1201 catacctgat ggagcctgca tctgaggatg attcattgtg ctcacacgcc aggcccctgt
     1261 gttactacta ggcagctcat tattctgcac tgaaccggag tttggtaaac tagaggtcat
     1321 aaagttagta gtgttatttg gtccagtcat ttcagtggta atattttgag gttgaccact
     1381 gaatgaaacc ttctgtgtac cacttacttc agattcacaa gtttcctgaa ggcttcccca
     1441 ggtaccatgg gtagaaccac cactcacttt agagttaata ctctgattgt taggcatctg
     1501 gcctatagta ctgcactgaa tattaatgcc agaactacca ctccctacag gcccttttag
     1561 ggcaagtcca ttgttctcta atactggcca ggcaccatgg ttgctagctg aattcaaagt
     1621 gcttggattt aaccctccat ttgatgaaga acttacagtg ccccaagcat tcattctatt
     1681 gttgctactt tcagatttgc tttcaggagc atccacagag acctgacatg tgcttattat
     1741 ggctccatgg gaaaaacccc atggcccagt actacttcca tggcccacat tattgctgct
     1801 gctaccaact acaaacttgt tttgggaacc aagtccagtg ctattccgaa ggccatcttt
     1861 ttcaccacct gtgttccctg aagccatgat agtgatgttt ctctctgatt cagagctgga
     1921 ggcagaatca gcatccatag ctcgagatct gagtccggac ttgtacagct cgtccatgcc
     1981 gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt
     2041 ggggtctttg ctcagggcgg actgggtgct caggtagtgg ttgtcgggca gcagcacggg
     2101 gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc
     2161 gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt cggccatgat
     2221 atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc
     2281 cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac
     2341 ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta
     2401 gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct
     2461 gaagcactgc acgccgtagg tcagggtggt cacgagggtg ggccagggca cgggcagctt
     2521 gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcgccctc
     2581 gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac
     2641 caccccggtg aacagctcct cgcccttgct caccatggtg gcgaccggta gcgctagagc
     2701 ttgggtctcc ctatagtgag tcgtattaat ttcgataagc cagtaagcag tgggttctct
     2761 agttagccag agagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg
     2821 cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa
     2881 caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta
     2941 tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac
     3001 gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc
     3061 gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg
     3121 tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc
     3181 tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg
     3241 cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct
     3301 atgaactaat gaccccgtaa ttgattacta ttaataacta atgcatggcg gtaatacggt
     3361 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3421 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3481 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3541 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3601 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3661 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3721 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3781 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3841 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3901 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3961 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4021 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4081 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4141 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac
     4201 ctgaggctat ggcagggcct gccgccccga cgttggctgc gagccctggg ccttcacccg
     4261 aacttggggg gtggggtggg gaaaaggaag aaacgcgggc gtattggccc caatggggtc
     4321 tcggtggggt atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg
     4381 acccaacacc gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg
     4441 gtattgtctc cttccgtgtt tcagttagcc tccccctagg gtgggcgaag aactccagca
     4501 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
     4561 acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt
     4621 ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa
     4681 ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca
     4741 ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc
     4801 cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat
     4861 attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc
     4921 cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc
     4981 ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg
     5041 gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat
     5101 gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc
     5161 gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg
     5221 aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt cattcagggc
     5281 accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac
     5341 ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac
     5401 ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca
     5461 tcctgtctct tgatcgatct ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
     5521 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc
     5581 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
     5641 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     5701 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac
     5761 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct
     5821 ggttctttcc gcctcaggac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5881 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5941 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
     6001 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     6061 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     6121 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     6181 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     6241 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     6301 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     6361 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     6421 aatttacgcg ttaagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     6481 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     6541 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     6601 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     6661 gttatctaga tccggtggat cccgggcccg cggtaccgtc gactgcagtt aatcaagttg
     6721 acgagattgt tgcatcattt gctgctgcac actaagaggt cgaccctgct ggtcttgctg
     6781 cggccggttc ccagaaggca cgcttctctg actctgcgcc ctttgctgct gcgctaacaa
     6841 tcgctgtaac tgggagatct gagaaagctg gtttagctgg gatagctggt tcagcatggc
     6901 tacctgttga gggccaaaga gtggattcag gccaccgttg tttggtgcat acttcagcaa
     6961 tgaaactgga acctgagggg agagtaatgg aggaggcact tgagcacgga gattaggctg
     7021 agatgaacta agaggttgtg ctggaggctg ctgcatgccc cggggttgtg ctgctgtgtt
     7081 tccaacacca aacatactgg gattgccatt ctgtctaaca gaattcaagt tttgcatacc
     7141 cagccctgct atggagccaa gggcctggtt aggtagtgca ctatttgaag ggggaagctt
     7201 catgctttga ctggacataa actgcatgtt ttgggattca tctgctacaa tgccatcctt
     7261 atcaaaataa ggattgcgct ccatggaaga ctctttggaa atctgaggcc gagaaccagg
     7321 gcctttcccg accgttcgat tgtaatcacc aatatttagg ctatgtttat ctatctccat
     7381 tcgtttgtct tgtaacattc ctccagaaag gtccatctta ttgctttgta cattttcctc
     7441 tggtgagtct ctcgaaaaac tgatgtttga aaactgttta acaaatggat ttatccacgc
     7501 ttcttcttgt ttgtttccac ctttcatcat tcccctttcc cttctccttt ttttgccact
     7561 cagacccttc gatgacattt tctttgtata tggtggccaa tttaaagatg agttaagctc
     7621 ttgagatgaa ttactattcc acattccaat ctccacatcc tcttcctctt cccagccagt
     7681 ggggcgattt ccaggcaatg gcatatcatc accacaccag ccatcttgca tagattttgg
     7741 cccagatttg cttaatgctt gtggaccaac agagctggag ccccacgtgg atgtgctgga
     7801 tgccgcagca atgggttccc cccagctggg accactgtct atgggcttac cccatgctga
     7861 agtaccatta tccacagttg tggctggagt agaaggctcc ccccagggct caccccagcc
     7921 agagccactg cttgcctctt tgtttgagat ggcacttgca ggcgttagca gctgacgtac
     7981 ctgtgcttgc tggtctgaac ggctgttgcc atttgggacg tttttgttcc acatgttcac
     8041 atttttgtag ttgtatttgc ttggatctcc ccaagctgaa gttccatcat caatctccat
     8101 tttgcgacgt atagattctg gggatggttc ctcccagcct gtgggttctt cttcttttgc
     8161 tggggctggt ataggtcccc ccagccagcc tgtaccagga ggtttgcctg tagctggtgg
     8221 gattccccaa gatccaacaa tgtcttgttg cttgttccag tctggagagc tgactggctt
     8281 tgacgaatct ccccaaccta gagactgatt agactttgga gggtctcccc atccctggga
     8341 aggagtaggt ttagaagatt catcccatcc tgatgaatta tttggattta tgttgtt
//
