LOCUS       sequence-426350-        6874 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HseIF4AII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6874)
  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 6874)
  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: pCIneo-V5-SBP-HseIF4AII_AC
FEATURES             Location/Qualifiers
     source          1..6874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     589..613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          753..885
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     868..887
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        930..948
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      961..970
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             970..1011
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1033..1146
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWGGGHVVEGLAGELEQLRARLEHHPQGQREP"
     polyA_signal    2441..2562
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2452..2471)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2506..2525
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2748..3203
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2835..2854)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3045..3066
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3320..3677
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3320..3511
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3343..3363)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3528..3663
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3728..4522
                     /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(3782..3801)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4392..4411
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4586..4634
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4675..4694)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4794..4816
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4854..4872)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4940..5044
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5045..5113,5114..5905)
                     /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(5263..5282)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6076..6664
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6565..6584
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcattgatt 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 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      601 gaaccgtcag atcactagaa gctttattgc ggtagtttat cacagttaaa ttgctaacgc
      661 agtcagtgct tctgacacaa cagtctcgaa cttaagctgc agtgactctc ttaaggtagc
      721 cttgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
      781 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
      841 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
      901 tcaattacag ctcttaaggc tagagtactt aatacgactc actataggct agcaattgcg
      961 gccaccatgg gtaagcctat ccctaaccct ctcctcggtc tcgattctac gcccggaagc
     1021 ggtagcggta ccatggacga gaagaccacc ggctgggggg gcggccacgt ggtggagggc
     1081 ctggccggcg agctggagca gctgcgggcc aggctggagc accaccctca gggccagcgg
     1141 gagccctccg gcggctgcaa gcccctcgag atgtctggtg gctccgcgga ttataacaga
     1201 gaacatggcg gcccagaggg aatggacccc gatggtgtca tcgagagcaa ctggaatgag
     1261 attgttgata actttgatga tatgaattta aaggagtctc tccttcgtgg catctatgct
     1321 tacggttttg agaagccttc cgctattcag cagagagcta ttattccctg tattaaaggg
     1381 tatgatgtga ttgctcaagc tcagtcaggt actggcaaga cagccacatt tgctatttcc
     1441 atcctgcaac agttggagat tgagttcaag gagacccaag cactagtatt ggcccccacc
     1501 agagaactgg ctcaacagat ccaaaaggta attctggcac ttggagacta tatgggagcc
     1561 acttgtcatg cctgcattgg tggaacaaat gttcgaaatg aaatgcaaaa actgcaggct
     1621 gaagcaccac atattgttgt tggtacaccc gggagagtgt ttgatatgtt aaacagaaga
     1681 tacctttctc caaaatggat caaaatgttt gttttggatg aagcagatga aatgttgagc
     1741 cgtggtttta aggatcaaat ctatgagatt ttccaaaaac taaacacaag tattcaggtt
     1801 gtgttgcttt ctgccacaat gccaactgat gtgttggaag tgaccaaaaa attcatgaga
     1861 gatccaattc gaattctggt gaaaaaggaa gaattgaccc ttgaaggaat caaacagttt
     1921 tatattaatg ttgagagaga ggaatggaag ttggatacac tttgtgactt gtacgagaca
     1981 ctgaccatta cacaggctgt tatttttctc aatacgaggc gcaaggtgga ctggctgact
     2041 gagaagatgc atgccagaga cttcacagtt tctgctctgc atggtgacat ggaccagaag
     2101 gagagagatg ttatcatgag ggaattccgg tcagggtcaa gtcgtgttct gatcactact
     2161 gacttgttgg ctcgcgggat tgatgtgcaa caagtgtctt tggttataaa ttatgatcta
     2221 cctaccaatc gtgaaaacta tattcacaga attggcagag ggggtcgatt tgggaggaaa
     2281 ggtgtggcta taaactttgt tactgaagaa gacaagagga ttcttcgtga cattgagact
     2341 ttctacaata ctacagtgga ggagatgccc atgaatgtgg ctgaccttat ttaagcggcc
     2401 gcttcccttt agtgagggtt aatgcttcga gcagacatga taagatacat tgatgagttt
     2461 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2521 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     2581 cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc
     2641 tacaaatgtg gtaaaatccg ataaggatcg atccgggctg gcgtaatagc gaagaggccc
     2701 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag
     2761 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2821 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2881 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2941 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3001 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3061 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3121 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3181 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     3241 atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc tctgaaagag
     3301 gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     3361 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3421 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3481 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3541 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3601 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3661 gcctaggctt ttgcaaaaag cttgattctt ctgacacaac agtctcgaac ttaaggctag
     3721 agccaccatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3781 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3841 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3901 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3961 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4021 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4081 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4141 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4201 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4261 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4321 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4381 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4441 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4501 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4561 cccaacctgc catcacgatg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     4621 tggttttttg tgtgaatcga tagcgataag gatccgcgta tggtgcactc tcagtacaat
     4681 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4741 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4801 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     4861 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4921 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4981 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5041 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5101 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5161 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5221 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5281 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5341 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5401 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5461 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5521 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5581 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5641 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5701 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5761 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5821 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5881 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5941 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6001 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6061 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6121 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6181 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6241 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6301 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6361 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6421 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6481 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6541 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6601 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6661 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6721 catggctcga cagatcttca atattggcca ttagccatat tattcattgg ttatatagca
     6781 taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt
     6841 tatattggct catgtccaat atgaccgcca tgtt
//
