LOCUS       sequence-208934-        5124 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of humanized MS2 (V75E/A81G), fused to mVenus 
            in mammalian cells, under a hUBC promoter, V5 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5124)
  AUTHORS   Maass PG, Barutcu AR, Shechner DM, Weiner CL, Mele M, Rinn JL
  TITLE     Spatiotemporal allele organization by allele-specific CRISPR 
            live-cell imaging (SNP-CLING).
  JOURNAL   Nat Struct Mol Biol. 2018 Jan 8. pii: 10.1038/s41594-017-0015-3. 
            doi: 10.1038/s41594-017-0015-3.
  PUBMED    29343869
REFERENCE   2  (bases 1 to 5124)
  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"; journalName: "Nat Struct 
            Mol Biol. 2018 Jan 8. pii: 10.1038/s41594-017-0015-3. doi: 
            10.1038/s41594-017-0015-3."
COMMENT     Sequence Label: pUBC_V5_MS2-mVenus
FEATURES             Location/Qualifiers
     source          1..5124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(1..792,793..861))
                     /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     624..643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(862..966)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1056..1075
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1269..1291
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1283..1300
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1284..1300
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1329..2540
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2377..2396
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      2558..2567
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2570..2590
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2597..2638
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             join(3068..3070,3071..3073,3074..3787)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3113..3134)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3374..3393)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3721..3742
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3834..3853)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3888..3907
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3964..3980)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3964..3980)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3977..3999)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3988..4004
                     /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(4012..4018,4019..4036,4037..4042))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4057..4078
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4195..4212)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4366..4954)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4446..4465)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
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 ctatgcggca tcagagcaga ttgtactgag
     1081 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     1141 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     1201 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     1261 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc tcggtacacg
     1321 tatgtcgagg taggcgcgcc gggttttggc gcctcccgcg ggcgcccccc tcctcacggc
     1381 gagcgctgcc acgtcagacg aagggcgcag cgagcgtcct gatccttccg cccggacgct
     1441 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga
     1501 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac
     1561 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga
     1621 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg
     1681 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct
     1741 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga
     1801 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg
     1861 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg
     1921 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt
     1981 gaggccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg gcaccatctg
     2041 gggaccctga cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg
     2101 gcggcagtta tggcggtgcc gttgggcagt gcacccgtac ctttgggagc gcgcgccctc
     2161 gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc agggtggggc cacctgccgg
     2221 taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc
     2281 tcctgaatcg acaggcgccg gacctctggt gaggggaggg ataagtgagg cgtcagtttc
     2341 tttggtcggt tttatgtacc tatcttctta agtagctgaa gctccggttt tgaactatgc
     2401 gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat
     2461 ttgggtcaat atgtaatttt cagtgttaga ctagtaaatt gtccgctaaa ttctggccgt
     2521 ttttggcttt tttgttagac ctggaggtac cgccaccgcc accatggctc caaagaagaa
     2581 gcgtaaggta ggcagcggca agcccatccc caaccccctg ctgggcctgg acagcaccgg
     2641 cagcgctagc aatttcactc aatttgtgct cgtggataac ggaggaacgg gcgacgtgac
     2701 cgtggcacca agcaactttg ctaatggagt ggccgagtgg attagctcaa atagcaggtc
     2761 ccaggcttac aaggtcacct gtagtgtccg gcaatcatcc gcccagaatc ggaaatatac
     2821 tattaaggta gaagtgccga aagtggcaac tcaaaccgtg gggggcgagg aattgcctgt
     2881 ggccggctgg agaagctacc tcaacatgga actcaccatt ccaatcttcg ccacaaacag
     2941 cgactgtgag ctcatcgtca aagccatgca gggtctgctg aaggacggca atcctattcc
     3001 atctgcaatc gcggccaatt ccggcattta tggatccaag ggctcgggct cgacctcggg
     3061 ctccggaatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     3121 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     3181 cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg
     3241 gcccaccctc gtgaccaccc tgggctacgg cctgcagtgc ttcgcccgct accccgacca
     3301 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     3361 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     3421 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     3481 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca
     3541 gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca
     3601 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     3661 caaccactac ctgagctacc agtccaagct gagcaaagac cccaacgaga agcgcgatca
     3721 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     3781 caagtaatga taagtcgact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3841 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3901 actcatcaat gtatcttatt cgactgttta aacctgcagg catgcaagct tggcgtaatc
     3961 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4021 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4081 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4141 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4201 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4261 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4321 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4381 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4441 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4501 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4561 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4621 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4681 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4741 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4801 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4861 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4921 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4981 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5041 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5101 atatgagtaa acttggtctg acag
//
