LOCUS       pUBC_V5_MS2~mChe        5255 bp    DNA     circular UNA 07-JUL-2015
DEFINITION  Transient expression of humanized MS2 (V75E/A81G), fused to mCherry
            in human cells, under a UBC promoter.
ACCESSION   urn.local...1435874812996.48
VERSION     urn.local...1435874812996.48
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     gene            complement(1..861)
                     /gene="bla"
                     /label="bla gene"
     CDS             complement(1..861)
                     /gene="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /codon_start=1
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /label="bla CDS"
     sig_peptide     complement(793..861)
                     /gene="bla"
                     /note="required for secretion to the periplasm; cleaved
                     off to form the mature beta-lactamase protein."
                     /label="bla signal peptide"
     -10_signal      complement(905..910)
                     /note="bla promoter (counter-clockwise); GAGACA;
                     transcript start 2548 (complementary strand)"
                     /label="-10 signal"
     -35_signal      complement(926..931)
                     /note="bla promoter (counter-clockwise); TTCAAA"
                     /label="-35 signal"
     source          order(<1058..>1452,<4051..>1057)
                     /organism="Cloning vector pNEB193"
                     /mol_type="other DNA"
                     /label="source Cloning vector pNEB193"
     primer_bind     1401..1424
                     /created_by="User"
                     /label="M13F (-47) Primer from GeneWiz"
     primer_bind     1427..1444
                     /created_by="User"
                     /label="M13F(-21) Primer From GeneWiz"
     Promoter        1475..2679
                     /created_by="User"
                     /label="human_UbC_Promoter"
     misc_feature    2698..2707
                     /created_by="User"
                     /label="Kozak_Sequence"
     sig_peptide     2704..2730
                     /vntifkey=21
                     /modified_by="User"
                     /label="NLS"
     CDS             2704..3171
                     /created_by="User"
                     /label="NLS_V5_MS2 (V75E/A81G)"
     misc_feature    2737..2778
                     /created_by="User"
                     /label="V5_Tag"
     CDS             3172..3207
                     /created_by="User"
                     /label="Linker"
     CDS             3208..3915
                     /created_by="User"
                     /label="mCherry"
     terminator      3931..4050
                     /label="SV40_PA_terminator"
     source          <3931..>4050
                     /organism="pcDNA3.1(+)"
                     /mol_type="other DNA"
                     /label="source pcDNA3.1(+)"
     misc_feature    complement(3965..3984)
                     /created_by="User"
                     /modified_by="User"
                     /label="SV40pA Rev Primer from Genewiz"
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     4019..4038
                     /modified_by="User"
                     /label="EBV_rev_primer"
     misc_feature    complement(4095..4111)
                     /created_by="User"
                     /label="M13R Primer From GeneWiz"
                     /note="Geneious type: primer_bind_reverse"
     -10_signal      complement(4144..4149)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TATGTT"
                     /label="-10 signal"
     -35_signal      complement(4168..4173)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TTTACA"
                     /label="-35 signal"
     protein_bind    complement(4193..4205)
                     /note="CAP protein binding site"
                     /bound_moiety="catabolite activator protein (CAP)"
                     /label="CAP protein binding site protein bind"
     rep_origin      complement(4497..5085)
                     /note="pMB1 origin of replication (counter-clockwise)
                     (RNAII -35 to RNA/DNA switch point)"
                     /label="rep origin"
     misc_RNA        complement(4497..5049)
                     /note="RNAII transcript (complementary strand)"
                     /label="RNA"
     -35_signal      4903..4908
                     /note="RNAI promoter (clockwise); TTGAAG"
                     /label="RNAI promoter (clockwise); TTGAAG -35 signal"
     -10_signal      4925..4930
                     /note="RNAI promoter (clockwise); GCTACA"
                     /label="RNAI promoter (clockwise); GCTACA -10 signal"
     misc_RNA        4939..5046
                     /note="RNAI transcript"
                     /label="RNAI transcript"
     -10_signal      complement(5059..5064)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
                     /label="-10 signal"
     -35_signal      complement(5080..5085)
                     /note="RNAII promoter (counter-clockwise); TTGAGA"
                     /label="-35 signal"
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 tgacgtctaa gaaaccatta ttatcatgac
     1021 attaacctat aaaaataggc gtatcacgag gccctttgcg cgtttcggtg atgacggtga
     1081 aaacctctga cacatgcagc tcccggtcac agcttgtctg taagcggatg ccgggagcag
     1141 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     1201 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     1261 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     1321 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     1381 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     1441 cagtgaattc gagctcggta cacgtatgtc gagggcgccg ggttttggcg cctcccgcgg
     1501 gcgcccccct cctcacggcg agcgctgcca cgtcagacga agggcgcagc gagcgtcctg
     1561 atccttccgc ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac
     1621 cccagtatca gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt
     1681 tctttccaga gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg
     1741 gatctccgtg gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc
     1801 tagttccgtc gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc
     1861 acttggtgag tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg
     1921 tgggacggaa gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt
     1981 gccctgaact gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat
     2041 ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc
     2101 ggcaagaacc caaggtcttg aggccttcgc taatgcggga aagctcttat tcgggtgaga
     2161 tgggctgggg caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg
     2221 tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc
     2281 tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca
     2341 gggtggggcc acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt
     2401 tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga
     2461 taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag
     2521 ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca
     2581 ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac tagtaaattg
     2641 tccgctaaat tctggccgtt tttggctttt ttgttagacc ctggaggtac cgccaccgcc
     2701 accatggctc caaagaagaa gcgtaaggta ggcagcggca agcccatccc caaccccctg
     2761 ctgggcctgg acagcaccgg cagcgctagc aatttcactc aatttgtgct cgtggataac
     2821 ggaggaacgg gcgacgtgac cgtggcacca agcaactttg ctaatggagt ggccgagtgg
     2881 attagctcaa atagcaggtc ccaggcttac aaggtcacct gtagtgtccg gcaatcatcc
     2941 gcccagaatc ggaaatatac tattaaggta gaagtgccga aagtggcaac tcaaaccgtg
     3001 gggggcgagg aattgcctgt ggccggctgg agaagctacc tcaacatgga actcaccatt
     3061 ccaatcttcg ccacaaacag cgactgtgag ctcatcgtca aagccatgca gggtctgctg
     3121 aaggacggca atcctattcc atctgcaatc gcggccaatt ccggcattta tggatccaag
     3181 ggctcgggct cgacctcggg ctccggaatg gtgagcaagg gcgaggagga caacatggcc
     3241 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccgcgag
     3301 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     3361 aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     3421 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     3481 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     3541 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     3601 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     3661 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     3721 ctgaaggacg gcggccacta cgacgccgag gtcaagacca cctacaaggc caagaagccc
     3781 gtgcagctgc ccggcgccta caacgtcaac atcaagctgg acatcacctc ccacaacgag
     3841 gactacacca tcgtggaaca gtacgagcgc gccgagggcc gccactccac cggcggcatg
     3901 gacgagctgt acaagtaatg ataagtcgac ttgtttattg cagcttataa tggttacaaa
     3961 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4021 ggtttgtcca aactcatcaa tgtatcttat tcgactgttt aaacctgcag gcatgcaagc
     4081 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4141 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4201 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4261 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4321 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4381 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4441 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4501 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4561 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4621 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4681 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4741 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4801 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4861 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4921 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4981 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5041 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5101 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5161 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5221 atctaaagta tatatgagta aacttggtct gacag
//
