LOCUS       sequence-181324-        6199 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses V5-PPP6C, includes IRES-Emerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6199)
  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 6199)
  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: pCAGIG_V5-PPP6C
FEATURES             Location/Qualifiers
     source          1..6199
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     287..309
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     371..390
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             429..470
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    480..504
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1426..1450)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1527..2100
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1550..2102
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1694..1711)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1921..1940
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2114..2116,2117..2119,2120..2833)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2159..2180)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2420..2439)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2767..2788
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2906..2925)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2971..3026
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3025..3044)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3387..3403)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3387..3403)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3400..3422)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3411..3427
                     /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(3435..3441,3442..3459,3460..3465))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3480..3501
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3560..3755
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3606..3741
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3761..3895
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3798..3817)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3852..3871
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3963..3980)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4134..4722)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4214..4233)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4893..5684,5685..5753))
                     /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     5516..5535
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5754..5858)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag
       61 agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc
      121 cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
      181 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc
      241 tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg
      301 cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta
      361 cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattacca
      421 ccatggcagg caagccaatc cctaaccctc tgctgggcct ggacagcacc gaattgatca
      481 caagtttgta caaaaaagca ggcaccatgg cgccgctaga cctggacaag tatgtggaaa
      541 tagcgcggct gtgcaagtac ctgccagaga acgacctgaa gcggctatgt gactacgttt
      601 gtgacctcct cttagaagag tcaaatgttc agccagtatc aacaccagta acagtgtgtg
      661 gagatatcca tggacagttt tatgaccttt gtgaactgtt cagaactgga ggtcaggttc
      721 ctgacacaaa ctacatattt atgggtgatt ttgtagacag aggttactat agtttggaga
      781 ccttcactta ccttcttgca ttaaaggcta aatggcctga tcgtattaca cttttgcgag
      841 gaaatcatga gagtagacag ataacacagg tctatggatt ttatgatgag tgccaaacca
      901 aatatggaaa tgctaatgcc tggagatact gtaccaaagt ttttgacatg ctcacagtag
      961 cagctttaat agatgagcag attttgtgtg tccatggtgg tttatctcct gatatcaaaa
     1021 cactggatca aattcgaacc atcgaacgga atcaggaaat tcctcataaa ggagcatttt
     1081 gtgatctggt ttggtcagat cctgaagatg tggatacctg ggctatcagt ccccgaggag
     1141 caggttggct ttttggagca aaggtcacaa atgagtttgt tcatatcaac aacttaaaac
     1201 tcatctgcag agcacatcaa ctagtgcacg aaggctataa atttatgttt gatgagaagc
     1261 tggtgacagt atggtctgct cctaattact gctatcgttg tggaaatatt gcttcgatca
     1321 tggtcttcaa agatgtaaat acaagagaac caaagttatt ccgggcagtt ccagattcag
     1381 aacgtgttat tcctcccaga acgacaacgc catatttcct ttagaaccca gctttcttgt
     1441 acaaagtggt tgatctcgag gattggcgcg ccaagatatc ttaattaagt taacaggcgg
     1501 ccgctacgta aattccgccc cccccccccc tctccctccc ccccccctaa cgttactggc
     1561 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     1621 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     1681 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     1741 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     1801 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     1861 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     1921 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     1981 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     2041 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     2101 tatggccaca tcgatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2161 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     2221 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     2281 gccctggccc accctcgtga ccaccttcac ctacggcgtg cagtgcttcg cccgctaccc
     2341 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2401 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2461 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2521 catcctgggg cacaagctgg agtacaacta caacagccac aaggtctata tcaccgccga
     2581 caagcagaag aacggcatca aggtgaactt caagacccgc cacaacatcg aggacggcag
     2641 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2701 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     2761 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2821 gctgtacaag taacaattgg catgcggccg gccgccagca cagtggtcga tcgaccaatg
     2881 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca
     2941 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     3001 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     3061 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca
     3121 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt
     3181 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt
     3241 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc
     3301 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc
     3361 tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3421 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3481 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3541 ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc
     3601 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3661 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3721 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta
     3781 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3841 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg
     3901 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3961 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4021 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4081 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4141 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4201 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4261 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4321 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4381 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4441 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4501 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4561 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4621 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4681 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4741 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4801 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4861 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4921 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4981 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5041 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5101 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5161 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5221 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5281 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5341 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5401 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5461 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5521 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5581 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5641 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5701 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5761 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5821 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5881 cctggtcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     5941 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     6001 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     6061 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     6121 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     6181 cccgcctggc attatgccc
//
