LOCUS       sequence-207484-        5082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of non-aggregating delta-FG mutant, fused to 
            mCherry in mammalian cells, under a hUBC promoter, HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5082)
  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 5082)
  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_HA_PP7-mCherry
FEATURES             Location/Qualifiers
     source          1..5082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     728..747
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      909..918
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             921..941
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             948..974
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1386..2096
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGREFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1531..1549)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1799..1818
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1993..2016
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2143..2162)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2197..2216
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2273..2289)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2273..2289)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2286..2308)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2297..2313
                     /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(2321..2327,2328..2345,2346..2351))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2366..2387
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2504..2521)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2675..3263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2755..2774)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3434..4225,4226..4294))
                     /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     4057..4076
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4295..4399)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4489..4508
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4702..4724
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4716..4733
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4717..4733
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(4762..5082,1..891)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
ORIGIN
        1 gtgtggcaca gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc
       61 gctgtgatcg tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc
      121 cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc
      181 gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc
      241 cgagtcttga atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg
      301 ggcatggtgg gcggcaagaa cccaaggtct tgaggccttc gctaatgcgg gaaagctctt
      361 attcgggtga gatgggctgg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac
      421 tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag
      481 tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg
      541 gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc
      601 aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg
      661 tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt
      721 aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa
      781 gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag
      841 actagtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga cctggaggta
      901 ccgccaccgc caccatggct ccaaagaaga agcgtaaggt aggcagctac ccatacgatg
      961 ttccagatta cgctggcagc tccatgtcaa agacgatcgt gctgtctgta ggtgaagcca
     1021 cacgcaccct cactgagata cagtccacgg ccgataggca gatttttgag gagaaggtgg
     1081 gtccactggt gggtaggctt cgactgacgg cctccctgag gcaaaacggc gctaaaacgg
     1141 cttatcgcgt taacctcaag ctggaccagg ccgatgtcgt tgatagtgga ctgcccaaag
     1201 tcaggtacac acaggtgtgg agccacgatg tgaccatcgt ggctaactca acagaggcgt
     1261 cccggaagtc tctgtacgac ctgacaaagt ccctggttgc cacctctcag gtggaggatc
     1321 tggtggttaa tctcgtgccc ctcggacggg gatccaaggg ctcgggctcg acctcgggct
     1381 ccggaatggt gagcaagggc gaggaggaca acatggccat catcaaggag ttcatgcgct
     1441 tcaaggtgca catggagggc tccgtgaacg gccgcgagtt cgagatcgag ggcgagggcg
     1501 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc
     1561 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
     1621 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
     1681 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
     1741 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     1801 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
     1861 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     1921 acgccgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     1981 acgtcaacat caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     2041 acgagcgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaatgat
     2101 aagtcgactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2161 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2221 tatcttattc gactgtttaa acctgcaggc atgcaagctt ggcgtaatca tggtcatagc
     2281 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     2341 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     2401 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     2461 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2521 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2581 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2641 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2701 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2761 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2821 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2881 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2941 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3001 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3061 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3121 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3181 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3241 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3301 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3361 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3421 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3481 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3541 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3601 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3661 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3721 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3781 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3841 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3901 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3961 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4021 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4081 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4141 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4201 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4261 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4321 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4381 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     4441 ttattatcat gacattaacc tataaaaata ggcgtatcac gagctatgcg gcatcagagc
     4501 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     4561 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     4621 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     4681 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattcg
     4741 agctcggtac acgtatgtcg aggtaggcgc gccgggtttt ggcgcctccc gcgggcgccc
     4801 ccctcctcac ggcgagcgct gccacgtcag acgaagggcg cagcgagcgt cctgatcctt
     4861 ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt
     4921 atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc
     4981 cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc
     5041 cgtggggcgg tgaacgccga tgattatata aggacgcgcc gg
//
