LOCUS       sequence-181325-        5923 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses V5-CSNK1A1, includes IRES-mCherry .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5923)
  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 5923)
  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_V5-CSNK1A1
FEATURES             Location/Qualifiers
     source          1..5923
                     /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..255
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1273..1297)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    join(1374..1948,1949..1951,1952..1960)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1541..1558)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1768..1787
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1961..2671
                     /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(2106..2124)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2374..2393
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2738..2757)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2803..2858
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2857..2876)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3219..3235)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3219..3235)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3232..3254)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3243..3259
                     /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(3267..3273,3274..3291,3292..3297))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3312..3333
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3392..3587
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3438..3573
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3593..3727
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3630..3649)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3684..3703
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3795..3812)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3966..4554)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4046..4065)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4725..5516,5517..5585))
                     /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     5348..5367
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5586..5690)
                     /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 aggcaccatg gcgagtagca gcggctccaa ggctgaattc attgtcggag
      301 ggaaatataa actggtacgg aagatcgggt ctggctcctt cggggacatc tatttggcga
      361 tcaacatcac caacggcgag gaagtggcag tgaagctaga atctcagaag gccaggcatc
      421 cccagttgct gtacgagagc aagctctata agattcttca aggtggggtt ggcatccccc
      481 acatacggtg gtatggtcag gaaaaagact acaatgtact agtcatggat cttctgggac
      541 ctagcctcga agacctcttc aatttctgtt caagaaggtt cacaatgaaa actgtactta
      601 tgttagctga ccagatgatc agtagaattg aatatgtgca tacaaagaat tttatacaca
      661 gagacattaa accagataac ttcctaatgg gtattgggcg tcactgtaat aagttattcc
      721 ttattgattt tggtttggcc aaaaagtaca gagacaacag gacaaggcaa cacataccat
      781 acagagaaga taaaaacctc actggcactg cccgatatgc tagcatcaat gcacatcttg
      841 gtattgagca gagtcgccga gatgacatgg aatcattagg atatgttttg atgtatttta
      901 atagaaccag cctgccatgg caagggctaa aggctgcaac aaagaaacaa aaatatgaaa
      961 agattagtga aaagaagatg tccacgcctg ttgaagtttt atgtaagggg tttcctgcag
     1021 aatttgcgat gtacttaaac tattgtcgtg ggctacgctt tgaggaagcc ccagattaca
     1081 tgtatctgag gcagctattc cgcattcttt tcaggaccct gaaccatcaa tatgactaca
     1141 catttgattg gacaatgtta aagcagaaag cagcacagca ggcagcctct tccagtgggc
     1201 agggtcagca ggcccaaacc cccacaggca agcaaactga caaaaccaag agtaacatga
     1261 aaggtttcta gaacccagct ttcttgtaca aagtggttga tctcgaggat tggcgcgcca
     1321 agatatctta attaagttaa caggcggccg ctacgtaaat tccgcccccc ccccccctct
     1381 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     1441 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     1501 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     1561 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     1621 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     1681 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     1741 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     1801 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
     1861 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     1921 gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca agggcgagga
     1981 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     2041 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     2101 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     2161 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     2221 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     2281 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     2341 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     2401 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     2461 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     2521 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     2581 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     2641 caccggcggc atggacgagc tgtacaagtg agtttggggc cggccgccag cacagtggtc
     2701 gatcgaccaa tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt
     2761 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     2821 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     2881 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     2941 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     3001 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     3061 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     3121 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga
     3181 gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     3241 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3301 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3361 cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc
     3421 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3481 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3541 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt
     3601 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3661 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     3721 ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3781 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3841 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3901 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3961 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4021 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4081 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4141 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4201 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4261 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4321 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4381 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4441 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4501 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4561 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4621 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4681 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4741 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4801 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4861 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4921 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4981 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5041 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5101 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5161 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5221 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5281 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5341 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5401 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5461 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5521 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5581 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5641 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5701 cgaaaagtgc cacctggtcg acattgatta ttgactagtt attaatagta atcaattacg
     5761 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     5821 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     5881 atagtaacgc caatagggac tttccattga cgtcaatggg tgg
//
