LOCUS       sequence-289546-        6082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsRck_296-472.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6082)
  AUTHORS   Chen Y, Boland A, Kuzuoglu-Ozturk D, Bawankar P, Loh B, Chang CT, 
            Weichenrieder O, Izaurralde E
  TITLE     A DDX6-CNOT1 complex and W-binding pockets in CNOT9 reveal direct 
            links between miRNA target recognition and silencing.
  JOURNAL   Mol Cell. 2014 Jun 5;54(5):737-50. doi: 
            10.1016/j.molcel.2014.03.034. Epub 2014 Apr 24.
  PUBMED    24768540
REFERENCE   2  (bases 1 to 6082)
  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: 
            "10.1016/j.molcel.2014.03.034"; journalName: "Mol Cell"; date: 
            "2014-06-5- 5"; volume: "54"; issue: "5"; pages: "737-50"
COMMENT     Sequence Label: pCIneo-lambdaN-HA-HsRCK_296-472_O
FEATURES             Location/Qualifiers
     source          1..6082
                     /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             1176..1202
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1786..1907
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1797..1816)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1851..1870
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2093..2548
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2180..2199)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2390..2411
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2665..3022
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2665..2856
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2688..2708)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2873..3008
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3073..3867
                     /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(3127..3146)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3737..3756
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3931..3979
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4020..4039)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4139..4161
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4199..4217)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4285..4389
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4390..4458,4459..5250)
                     /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(4608..4627)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5421..6009
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5910..5929
                     /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 aggcttaccc atacgatgtt ccagattacg
     1201 ctcctaaggg agtaacccag tactacgcat atgtaactga gcgccaaaaa gtacactgcc
     1261 tcaacacact tttctccagg cttcagataa accagtcgat cattttctgt aactcctctc
     1321 agcgagttga attgctagcc aagaagattt ctcaactggg ttattcttgc ttctatattc
     1381 atgctaaaat gaggcaggaa catcgaaatc gtgtatttca tgatttccga aatggcttat
     1441 gccgcaatct tgtttgcact gatctgttta cccgaggtat tgatatacaa gctgtgaatg
     1501 tggtaataaa ctttgatttc ccaaagctgg cagagaccta tctccatcgt attggaagat
     1561 caggtcgctt tggtcatctt ggcttagcca tcaacttgat cacatatgat gatcgcttca
     1621 acctgaaaag tattgaggag cagctgggaa cagaaattaa acctattccg agcaacattg
     1681 ataagagcct gtatgtggca gaataccaca gcgagcctgt agaagatgag aaaccttaag
     1741 cggccgcttc cctttagtga gggttaatgc ttcgagcaga catgataaga tacattgatg
     1801 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     1861 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     1921 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa
     1981 acctctacaa atgtggtaaa atccgataag gatcgatccg ggctggcgta atagcgaaga
     2041 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc
     2101 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2161 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2221 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2281 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2341 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2401 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2461 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2521 tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     2581 gcggtatttc acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga
     2641 aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca
     2701 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2761 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2821 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2881 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     2941 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3001 tggaggccta ggcttttgca aaaagcttga ttcttctgac acaacagtct cgaacttaag
     3061 gctagagcca ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3121 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3181 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3241 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3301 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3361 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3421 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3481 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3541 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3601 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3661 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3721 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3781 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     3841 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     3901 cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg
     3961 tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc gcgtatggtg cactctcagt
     4021 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     4081 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4141 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4201 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4261 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4321 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4381 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4441 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4501 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4561 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4621 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4681 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4741 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4801 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4861 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4921 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4981 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5041 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5101 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5161 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5221 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5281 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5341 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5401 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5461 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5521 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5581 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5641 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5701 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5761 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5821 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5881 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5941 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6001 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6061 gctcacatgg ctcgacagat ct
//
