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
//
