LOCUS pCMV_mCherry_2A_ 5499 bp DNA circular UNA 07-JUL-2015 DEFINITION Transient expression of humanized MS2 (V75E/A81G), fused to the VP64 transcrption activator, in human cells, under a CMV promoter. ACCESSION urn.local...1435874543004.32 VERSION urn.local...1435874543004.32 KEYWORDS . SOURCE ORGANISM . FEATURES Location/Qualifiers gene complement(1..861) /gene="Ampicillin" /label="Ampicillin" CDS complement(1..861) /gene="Ampicillin" /note="ORF frame 1" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW*" /label="Ampicillin" promoter complement(903..931) /label="AmpR_promoter" origin 997^998 /label="origin" source 998..>1049 /organism="pcDNA3.1(+)" /mol_type="other DNA" /label="source pcDNA3.1(+)" source <1050..>1578 /organism="pcDNA3.1(+)" /mol_type="other DNA" /label="source pcDNA3.1(+)" rep_origin 1141..1447 /label="f1_origin" misc_feature complement(1561..1581) /modified_by="User" /label="pBABE_3_primer" /note="Geneious type: primer_bind_reverse" promoter 1582..2198 /label="CMV_immearly_promoter" source <1582..>2221 /organism="pcDNA3.1(+)" /mol_type="other DNA" /label="source pcDNA3.1(+)" misc_feature 1661..1948 /label="CAG_enhancer" primer_bind 2115..2135 /modified_by="User" /label="CMV_fwd_primer" misc_feature 2225..2234 /created_by="User" /label="Kozak_Sequence" sig_peptide 2231..2257 /vntifkey=21 /modified_by="User" /label="NLS" CDS 2258..2965 /created_by="User" /label="mCherry" CDS 2966..2980 /created_by="User" /modified_by="User" /label="Linker" CDS 2981..3043 /created_by="User" /modified_by="User" /label="2A_Self-Cleaving_Peptide" CDS 3044..3049 /created_by="User" /label="Linker" misc_feature 3050..3076 /created_by="User" /label="HA_Tag" CDS 3050..3478 /created_by="User" /label="HA_MS2_1" misc_feature 3314..3316 /created_by="User" /modified_by="User" /modified_by="User" /label="V75E" misc_feature 3332..3334 /created_by="User" /modified_by="User" /label="A81G" CDS 3479..3499 /created_by="User" /label="Linker" CDS 3500..3925 /created_by="User" /label="MS2_2_NLS" misc_feature 3719..3721 /created_by="User" /modified_by="User" /label="V75E" misc_feature 3737..3739 /created_by="User" /modified_by="User" /label="A81G" sig_peptide 3893..3925 /created_by="User" /modified_by="User" /modified_by="User" /label="NLS1" CDS 3926..4099 /created_by="User" /label="VP64 Transcription Activator" source <4114..>4132 /organism="pcDNA3.1(+)" /mol_type="other DNA" /label="source pcDNA3.1(+)" terminator 4133..4252 /label="SV40_PA_terminator" source <4133..>4525 /organism="pcDNA3.1(+)" /mol_type="other DNA" /label="source pcDNA3.1(+)" misc_feature complement(4167..4186) /created_by="User" /modified_by="User" /label="SV40pA Rev Primer from Genewiz" /note="Geneious type: primer_bind_reverse" primer_bind 4221..4240 /modified_by="User" /label="EBV_rev_primer" misc_feature complement(4296..4314) /modified_by="User" /label="M13_reverse_primer" /note="Geneious type: primer_bind_reverse" misc_feature complement(4313..4335) /modified_by="User" /label="M13_pUC_rev_primer" /note="Geneious type: primer_bind_reverse" promoter complement(4349..4378) /label="lac_promoter" terminator complement(4529..4682) /label="up_PA_terminator" source complement(<4529..>4682) /organism="pGL3-Basic" /mol_type="other DNA" /label="source pGL3-Basic" source <4684..997 /organism="pcDNA3.1(+)" /mol_type="other DNA" /label="source pcDNA3.1(+)" rep_origin complement(4729..5345) /label="pBR322_origin" ORIGIN 1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 961 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat 1021 cccctatggt gcactctcag tacaatctgg catgctgggg atgcggtggg ctctatggct 1081 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc 1141 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 1201 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 1261 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 1321 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 1381 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 1441 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt 1501 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt 1561 ggaatgtgtg tcagttaggg tacattgatt attgactagt tattaatagt aatcaattac 1621 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg 1681 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc 1741 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac 1801 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa 1861 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac 1921 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta 1981 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga 2041 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa 2101 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag 2161 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatag acccaagctg 2221 gctagccacc atggctccaa agaagaagcg taaggtaatg gtgagcaagg gcgaggagga 2281 caacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa 2341 cggccgcgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac 2401 cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc 2461 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt 2521 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg 2581 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa 2641 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg 2701 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca 2761 gaggctgaag ctgaaggacg gcggccacta cgacgccgag gtcaagacca cctacaaggc 2821 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagctgg acatcacctc 2881 ccacaacgag gactacacca tcgtggaaca gtacgagcgc gccgagggcc gccactccac 2941 cggcggcatg gacgagctgt acaagggcag caccagcggc ggcagtggag agggcagagg 3001 aagtctgcta acatgcggtg acgtcgagga gaatcctggc ccaggcagtt acccctacga 3061 cgtgcccgac tacgccggat ccggcgccgg taccagcaat ttcactcaat ttgtgctcgt 3121 ggataacgga ggaacgggcg acgtgaccgt ggcaccaagc aactttgcta atggagtggc 3181 cgagtggatt agctcaaata gcaggtccca ggcttacaag gtcacctgta gtgtccggca 3241 atcatccgcc cagaatcgga aatatactat taaggtagaa gtgccgaaag tggcaactca 3301 aaccgtgggg ggcgaggaat tgcctgtggc cggctggaga agctacctca acatggaact 3361 caccattcca atcttcgcca caaacagcga ctgtgagctc atcgtcaaag ccatgcaggg 3421 tctgctgaag gacggcaatc ctattccatc tgcaatcgcg gccaattccg gcatttatgg 3481 ttccacgagc gggtcaggga gcaatttcac tcaatttgtg ctcgtggata acggaggaac 3541 gggcgacgtg accgtggcac caagcaactt tgctaatgga gtggccgagt ggattagctc 3601 aaatagcagg tcccaggctt acaaggtcac ctgtagtgtc cggcaatcat ccgcccagaa 3661 tcggaaatat actattaagg tagaagtgcc gaaagtggca actcaaaccg tggggggcga 3721 ggaattgcct gtggccggct ggagaagcta cctcaacatg gaactcacca ttccaatctt 3781 cgccacaaac agcgactgtg agctcatcgt caaagccatg cagggtctgc tgaaggacgg 3841 caatcctatt ccatctgcaa tcgcggccaa ttccggcatt tatggttccg gggaccccaa 3901 gaagaagaga aaggtggagg ccagcggttc cggacgggct gacgcattgg acgattttga 3961 tctggatatg ctgggaagtg acgccctcga tgattttgac cttgacatgc ttggttcgga 4021 tgcccttgat gactttgacc tcgacatgct cggcagtgac gcccttgatg atttcgacct 4081 ggacatgctg attaactgaa taaatgaggg cccgtttaaa cccgctgatc agttgtttat 4141 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 4201 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 4261 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 4321 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 4381 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 4441 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 4501 cggtttgcgt attgggcgct cttccgctag agaaatgttc tggcacctgc acttgcactg 4561 gggacagcct attttgctag tttgttttgt ttcgttttgt tttgatggag agcgtatgtt 4621 agtactatcg attcacacaa aaaaccaaca cacagatgta atgaaaataa agatatttta 4681 ttacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 4741 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 4801 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 4861 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 4921 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 4981 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 5041 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 5101 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 5161 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 5221 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 5281 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 5341 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 5401 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 5461 atcaatctaa agtatatatg agtaaacttg gtctgacag //