LOCUS       sequence-289647-        6113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNanos3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6113)
  AUTHORS   Bhandari D, Raisch T, Weichenrieder O, Jonas S, Izaurralde E
  TITLE     Structural basis for the Nanos-mediated recruitment of the CCR4-NOT 
            complex and translational repression.
  JOURNAL   Genes Dev. 2014 Apr 15;28(8):888-901. doi: 10.1101/gad.237289.113.
  PUBMED    24736845
REFERENCE   2  (bases 1 to 6113)
  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: 
            "10.1101/gad.237289"; journalName: "Genes Dev"; date: "2014-04-15- 
            15"; volume: "28"; issue: "8"; pages: "888-901"
COMMENT     Sequence Label: pCIneo-lambdaN-HA-HsNanos3_P
FEATURES             Location/Qualifiers
     source          1..6113
                     /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      1098..1107
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1104..1169
                     /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             1194..1220
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1817..1938
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1828..1847)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1882..1901
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2124..2579
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2211..2230)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2421..2442
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2696..3053
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2696..2887
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2719..2739)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2904..3039
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3104..3898
                     /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(3158..3177)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3768..3787
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3962..4010
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4051..4070)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4170..4192
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4230..4248)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4316..4420
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4421..4489,4490..5281)
                     /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(4639..4658)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5452..6040
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5941..5960
                     /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 aattgcggcc accatggacg cacaaacacg acgacgtgag cgtcgcgctg
     1141 agaaacaagc tcaatggaaa gctgcaaacg gtggatctcc tccacatatg gcttacccat
     1201 acgatgttcc agattacgct cctccactcg agatgggcac cttcgacctg tggaccgact
     1261 atctgggact ggcccatctc gtgcgggccc tgtctggaaa agagggcccc gagacaagac
     1321 tgagccccca gcctgagcca gagcccatgc tggaacccga ccagaagcgg agcctggaaa
     1381 gctctcctgc ccccgagaga ctgtgcagct tctgcaagca caacggcgag agccgggcca
     1441 tctaccagag ccacgtgctg aaggatgagg ccggcagagt gctgtgcccc atcctgagag
     1501 actacgtgtg ccctcagtgc ggcgccacca gagagagagc ccacaccaga agattctgcc
     1561 ccctgaccgg ccagggctac accagcgtgt acagccacac caccagaaac agcgccggca
     1621 agaaactcgt gcggcccgac aaggccaaga cccaggatac cggacacaga agaggcggcg
     1681 gaggcggagc tggctttaga ggcgccggaa agtctgagcc cagccccagc tgtagcccca
     1741 gcatgagcac ctaatctaga gtcgacccgg gcggccgctt ccctttagtg agggttaatg
     1801 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     1861 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     1921 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     1981 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatccgataa
     2041 ggatcgatcc gggctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     2101 tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2161 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2221 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2281 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     2341 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2401 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     2461 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     2521 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc
     2581 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgcggatct
     2641 gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag
     2701 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     2761 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     2821 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2881 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2941 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     3001 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg
     3061 attcttctga cacaacagtc tcgaacttaa ggctagagcc accatgattg aacaagatgg
     3121 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3181 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3241 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     3301 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     3361 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     3421 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     3481 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     3541 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     3601 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     3661 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     3721 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     3781 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     3841 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     3901 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgatggccg
     3961 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagc
     4021 gataaggatc cgcgtatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4081 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4141 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4201 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4261 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4321 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4381 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4441 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4501 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4561 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4621 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4681 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4741 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4801 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4861 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4921 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4981 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5041 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5101 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5161 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5221 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5281 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5341 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5401 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5461 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5521 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5581 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5641 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5701 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5761 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5821 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5881 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5941 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6001 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6061 tttacggttc ctggcctttt gctggccttt tgctcacatg gctcgacaga tct
//
