LOCUS       sequence-289191-        6997 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsTNRC6Biso1_1250-1723-F1370A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6997)
  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 6997)
  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: pCIneo-lambdaN-HA-HsTNRC6Biso1_1250-1723-F1370A_K
FEATURES             Location/Qualifiers
     source          1..6997
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1089..1098
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1095..1160
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             1203..1229
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2701..2822
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2712..2731)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2766..2785
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3008..3463
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3095..3114)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3305..3326
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3580..3937
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3580..3771
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3603..3623)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3788..3923
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3988..4782
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4042..4061)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4652..4671
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4846..4894
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4935..4954)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5054..5076
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5114..5132)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5200..5304
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5305..5373,5374..6165)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5523..5542)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6336..6924
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6825..6844
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc caccatggac gcacaaacac gacgacgtga gcgtcgcgct gagaaacaag
     1141 ctcaatggaa agctgcaaac ccaccgctcg agaattcacg cgtggtacct ctagagatgg
     1201 cctacccata cgatgttcca gattacgctc cagaccttca aaccaaaggg ccaatacctg
     1261 gatatggttc tggcttcagc tctggcggca tggactatgg catggttggt gggaaggagg
     1321 ctggaaccga gtctcgcttt aaacagtgga cctccatgat ggaggggctg ccctctgtag
     1381 ccacacagga agccaatatg cacaaaaatg gcgctatagt ggcccctggt aaaacccggg
     1441 gagggtcacc gtacaaccag tttgatatca tccctggtga cacactgggt ggccatacgg
     1501 gtcctgctgg tgatagctgg ttacctgcca aatctccacc aacaaataaa atcggaagta
     1561 aatccagcaa tgccagttgg cctccagaag cccaaccagg agtgccatgg aaaggtatcc
     1621 aaaacattga ccctgaatct gacccctatg tcaccccagg aagtgtgctg gggggtacag
     1681 ccacatctcc cattgtagat actgaccacc aactgctgcg ggataacacc acagggtcta
     1741 attcttccct caacacctcg ctgccttcac ctggtgcctg gccctacagt gcctctgaca
     1801 actcctttac caacgttcat agcacttcag caaagttccc tgattacaaa tcaacatggt
     1861 ccccagatcc cataggacac aaccccactc atctctccaa caagatgtgg aaaaaccata
     1921 tttcctccag gaacactaca ccgctgcccc gcccacctcc tggtctgacc aaccccaaac
     1981 catcatctcc ctggagcagc acagcacccc gatcagtcag ggggtggggg acacaggact
     2041 cacggctcgc ctcggcctct acctggagtg atggtggctc agttcgtcct agttactggc
     2101 tggttcttca caatctcacc ccacagattg atgggtcaac cttgagaacg atctgcatgc
     2161 agcatggccc actgctgaca ttccatctga atctaaccca gggcactgcc ctgatccgat
     2221 acagcaccaa acaggaggcg gccaaggccc aaactgcact gcacatgtgt gtgttgggaa
     2281 acactaccat ccttgctgag tttgccactg atgatgaagt cagccgcttt ctggcacaag
     2341 ctcagccccc tacacctgca gcaaccccaa gtgcgccagc tgcggggtgg cagtcgctgg
     2401 agaccggcca gaaccagtca gatcccgtgg gacctgctct gaatcttttt ggtgggtcca
     2461 ctgggctcgg gcagtggagc agcagcgctg gtggcagcag cggggccgat cttgctggcg
     2521 cttcattgtg ggggccccca aactattctt ctagcttatg gggagtccca acggtggaag
     2581 atccccatag gatgggcagc cctgctcctt tactacctgg tgaccttctg ggaggagggt
     2641 cggattcaat ctgagcggcc gcttcccttt agtgagggtt aatgcttcga gcagacatga
     2701 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2761 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2821 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     2881 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatccg ataaggatcg atccgggctg
     2941 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     3001 cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3061 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3121 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3181 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3241 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3301 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3361 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3421 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
     3481 tccttacgca tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc
     3541 tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct
     3601 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3661 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3721 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3781 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3841 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3901 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttgattctt ctgacacaac
     3961 agtctcgaac ttaaggctag agccaccatg attgaacaag atggattgca cgcaggttct
     4021 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4081 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4141 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4201 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4261 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4321 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4381 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4441 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4501 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4561 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4621 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4681 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4741 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4801 aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa aatatcttta
     4861 ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag gatccgcgta
     4921 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4981 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     5041 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     5101 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5161 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5221 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5281 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5341 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5401 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5461 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5521 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5581 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5641 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5701 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5761 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5821 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5881 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5941 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6001 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6061 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6121 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6181 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6241 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6301 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6361 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6421 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6481 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6541 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6601 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6661 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6721 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6781 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6841 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6901 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6961 ttttgctggc cttttgctca catggctcga cagatct
//
