LOCUS       sequence-181327-        6561 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Empty backbone, includes IRES-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6561)
  AUTHORS   Golden RJ, Chen B, Li T, Braun J, Manjunath H, Chen X, Wu J, Schmid 
            V, Chang TC, Kopp F, Ramirez-Martinez A, Tagliabracci VS, Chen ZJ, 
            Xie Y, Mendell JT
  TITLE     An Argonaute phosphorylation cycle promotes microRNA-mediated 
            silencing.
  JOURNAL   Nature. 2017 Feb 9;542(7640):197-202. doi: 10.1038/nature21025. Epub
            2017 Jan 23.
  PUBMED    28114302
REFERENCE   2  (bases 1 to 6561)
  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/nature21025"; journalName: "Nature"; date: "2017-02-9- 9"; 
            volume: "542"; issue: "7640"; pages: "197-202"
COMMENT     Sequence Label: pCAGImC_Empty
FEATURES             Location/Qualifiers
     source          1..6561
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     38..60
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     122..141
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             180..221
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    231..355
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(380..385,386..403,404..410)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             464..1123
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(530..549)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1465..1770
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1649..1668
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1811..1935)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    join(2012..2586,2587..2589,2590..2598)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2179..2196)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2406..2425
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2599..3309
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2744..2762)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3012..3031
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3376..3395)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3441..3496
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3495..3514)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3857..3873)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3857..3873)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3870..3892)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3881..3897
                     /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(3905..3911,3912..3929,3930..3935))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3950..3971
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4030..4225
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4076..4211
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4231..4365
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4268..4287)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4322..4341
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4433..4450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4604..5192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4684..4703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5363..6154,6155..6223))
                     /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     5986..6005
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6224..6328)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
       61 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
      121 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattacc accatggcag
      181 gcaagccaat ccctaaccct ctgctgggcc tggacagcac cgaattgatc acaagtttgt
      241 acaaaaaagc tgaacgagaa acgtaaaatg atataaatat caatatatta aattagattt
      301 tgcataaaaa acagactaca taatactgta aaacacaaca tatccagtca ctatggcggc
      361 cgcattaggc accccaggct ttacacttta tgcttccggc tcgtataatg tgtggatttt
      421 gagttaggat ccgtcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac
      481 tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca
      541 gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa
      601 gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct
      661 gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg tgatatggga
      721 tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg
      781 gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg
      841 ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc
      901 agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggcaaata tggacaactt
      961 cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc
     1021 gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa
     1081 tgaattacaa cagtactgtg atgagtggca gggcggggcg taaacgcgtg gatccggctt
     1141 actaaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta taagaatata
     1201 tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca gcgtattaca
     1261 gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc aatatctccg
     1321 gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag
     1381 cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac ggctcttttg
     1441 ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa aagagagagc
     1501 cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg
     1561 gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg
     1621 gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg
     1681 gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac
     1741 gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca cagccagtct
     1801 gcaggtcgac catagtgact ggatatgttg tgttttacag tattatgtag tctgtttttt
     1861 atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc gttcagcttt
     1921 cttgtacaaa gtggttgatc tcgaggattg gcgcgccaag atatcttaat taagttaaca
     1981 ggcggccgct acgtaaattc cgcccccccc ccccctctcc ctcccccccc cctaacgtta
     2041 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
     2101 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     2161 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     2221 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     2281 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     2341 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     2401 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     2461 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt
     2521 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
     2581 gataatatgg ccacaaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
     2641 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
     2701 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     2761 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     2821 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
     2881 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     2941 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     3001 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     3061 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     3121 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     3181 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     3241 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     3301 tacaagtgag tttggggccg gccgccagca cagtggtcga tcgaccaatg ccctggctca
     3361 caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct
     3421 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     3481 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa
     3541 tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg
     3601 ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca
     3661 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc
     3721 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg
     3781 actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa
     3841 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3901 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3961 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4021 agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4081 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4141 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     4201 ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac
     4261 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4321 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa
     4381 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4441 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4501 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4561 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4621 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4681 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4741 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4801 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4861 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4921 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4981 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5041 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5101 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5161 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5221 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5281 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5341 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5401 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5461 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5521 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5581 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5641 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5701 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5761 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5821 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5881 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5941 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6001 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6061 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6121 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6181 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6241 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6301 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctggtcgac
     6361 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     6421 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     6481 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     6541 tccattgacg tcaatgggtg g
//
