LOCUS       sequence-474674-        7287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsRck.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7287)
  AUTHORS   Kuzuoglu-Ozturk D, Bhandari D, Huntzinger E, Fauser M, Helms S, 
            Izaurralde E
  TITLE     miRISC and the CCR4-NOT complex silence mRNA targets independently 
            of 43S ribosomal scanning.
  JOURNAL   EMBO J. 2016 Jun 1;35(11):1186-203. doi: 10.15252/embj.201592901. 
            Epub 2016 Mar 23.
  PUBMED    27009120
REFERENCE   2  (bases 1 to 7287)
  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: 
            "10.15252/embj.201592901"; journalName: "EMBO J"; date: "2016-06-1- 
            1"; volume: "35"; issue: "11"; pages: "1186-203"
COMMENT     Sequence Label: pcDNA3.1-MS2-HA-HsRck_AC
FEATURES             Location/Qualifiers
     source          1..7287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      661..670
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1099..1125
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2606..2624)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2607..2624)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2644..2661)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2650..2874
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2920..3348
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3007..3026)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3217..3238
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3357..3377)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3362..3691
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3542..3677
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3604..3623
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3758..4552
                     /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(3812..3831)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4422..4441
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4726..4847
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4763..4782)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4817..4836
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4896..4912)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4896..4912)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4909..4931)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4920..4936
                     /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)."
     promoter        complement(join(4944..4950,4951..4968,4969..4974))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4989..5010
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5127..5144)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5298..5886)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5378..5397)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6057..6848,6849..6917))
                     /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     6680..6699
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6918..7022)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7097..7116)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 gccaccatgg cttctaactt tactcagttc gttctcgtcg acaatggcgg aactggcgac
      721 gtgactgtcg ccccaagcaa cttcgctaac ggggtcgctg aatggatcag ctctaactcg
      781 cgatcacagg cttacaaagt aacctgtagc gttcgtcaga gctctgcgca gaatcgcaaa
      841 tacaccatca aagtcgaggt gcctaaagtg gcaacccaga ctgttggtgg tgaagagctt
      901 cctgtagccg gatggagatc ttacttaaat atggaactaa ccattccaat tttcgccacg
      961 aattccgact gcgagcttat tgttaaggca atgcaaggtc tcctaaaaga tggaaacccg
     1021 attccctcgg ccatcgcggc caactccggc atctacggag gtggaggtgg atctggtgga
     1081 tctcctccac atatggctta cccatacgat gttccagatt acgctcctcc aaagcttgga
     1141 tccgaattcc cgggtcgact cgagatgggt ctgtccagtc aaaatggtca gctgagaggc
     1201 cctgtgaaac ccactggtgg ccctggagga gggggcacac agacacagca acagatgaac
     1261 cagctgaaaa acaccaacac aatcaataat ggcactcagc agcaagcaca gagtatgacc
     1321 accactatta aacctggtga tgactggaaa aagactttaa aactccctcc aaaggatcta
     1381 agaatcaaaa cttcggatgt gacctccaca aaaggaaatg agtttgaaga ttactgtttg
     1441 aaacgggagt tactgatggg aatttttgaa atgggctggg aaaagccatc tcctattcag
     1501 gaggagagca ttcccattgc tttatctggt agggatatct tagctagagc aaaaaatgga
     1561 acaggcaaga gcggtgccta cctcattccc ttacttgaac ggctagacct gaagaaggac
     1621 aatatacaag caatggtgat tgttcccact agagaacttg ctctacaggt cagtcaaatt
     1681 tgcatccagg tcagcaaaca catgggaggg gccaaagtga tggcaaccac aggaggaacc
     1741 aatttacgag atgacataat gaggcttgat gatacagtgc acgtggtgat tgctacccct
     1801 gggagaatcc tggatcttat taagaaagga gtagcaaagg ttgatcatgt ccagatgata
     1861 gtattggatg aggcagataa gttgctgtca caggattttg tgcagataat ggaggatatt
     1921 attctcacgc tacctaaaaa caggcagatt ttactatatt ccgctacttt ccctcttagt
     1981 gtacagaagt tcatgaattc ccatttgcag aaaccctatg agattaacct gatggaggaa
     2041 ctaactctga agggagtaac ccagtactac gcatatgtaa ctgagcgcca aaaagtacac
     2101 tgcctcaaca cacttttctc caggcttcag ataaaccagt cgatcatttt ctgtaactcc
     2161 tctcagcgag ttgaattgct agccaagaag atttctcaac tgggttattc ttgcttctat
     2221 attcatgcta aaatgaggca ggaacatcga aatcgtgtat ttcatgattt ccgaaatggc
     2281 ttatgccgca atcttgtttg cactgatctg tttacccgag gtattgatat acaagctgtg
     2341 aatgtggtaa taaactttga tttcccaaag ctggcagaga cctatctcca tcgtattgga
     2401 agatcaggtc gctttggtca tcttggctta gccatcaact tgatcacata tgatgatcgc
     2461 ttcaacctga aaagtattga ggagcagctg ggaacagaaa ttaaacctat tccgagcaac
     2521 attgataaga gcctgtatgt ggcagaatac cacagcgagc ctgtagaaga tgagaaacct
     2581 taagcggccg cctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     2641 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2701 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2761 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2821 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     2881 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     2941 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3001 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3061 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3121 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3181 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3241 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3301 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     3361 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3421 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3481 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3541 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3601 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3661 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3721 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3781 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3841 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3901 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3961 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4021 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4081 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4141 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4201 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4261 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4321 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4381 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4441 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4501 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4561 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4621 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4681 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4741 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4801 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4861 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4921 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4981 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5041 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5101 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5161 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5221 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5281 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5341 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5401 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5461 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5521 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5581 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5641 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5701 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5761 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5821 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5881 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5941 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6001 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6061 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6121 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6181 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6241 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6301 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6361 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6421 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6481 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6541 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6601 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6661 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6721 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6781 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6841 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6901 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6961 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7021 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     7081 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     7141 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     7201 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     7261 gcgatgtacg ggccagatat acgcgtt
//
