LOCUS       sequence-205195-        4919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of nuclear localized mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4919)
  AUTHORS   Ariotti N, Rae J, Giles N, Martel N, Sierecki E, Gambin Y, Hall TE, 
            Parton RG
  TITLE     Ultrastructural localisation of protein interactions using 
            conditionally stable nanobodies.
  JOURNAL   PLoS Biol. 2018 Apr 5;16(4):e2005473. doi: 
            10.1371/journal.pbio.2005473. eCollection 2018 Apr.
  PUBMED    29621251
REFERENCE   2  (bases 1 to 4919)
  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: "PLoS 
            Biol."; date: "2018-04-5"; volume: "16(4):e2005473. doi"; pages: " 
            10.1371/journal.pbio.2005473. eCollection 2018 Apr"
COMMENT     Sequence Label: nls-mCherry
FEATURES             Location/Qualifiers
     source          1..4919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    108..132
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      136..145
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             148..168
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(173..197)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             199..906
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(341..359)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     609..628
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     803..826
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(908..928)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    968..1102
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(992..1011)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1046..1065
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1215..1235)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1215..1233)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1254..1270)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1254..1270)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1267..1289)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1278..1294
                     /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(1302..1308,1309..1326,1327..1332))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1347..1368
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1485..1502)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1656..2244)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1736..1755)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2415..3206,3207..3275))
                     /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     3038..3057
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3276..3380)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3406..3861)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3543..3564)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3755..3774
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3935..4919
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattca aggatcaaca agtttgtaca
      121 aaaaagcagg ctgccgccac catggctcca aagaagaagc gtaaggtacg gtacccagct
      181 ttcttgtaca aagtggctgt gagcaagggc gaggaggaca acatggccat catcaaggag
      241 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
      301 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
      361 ggcggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
      421 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
      481 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
      541 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
      601 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg
      661 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
      721 ggccactacg acgccgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
      781 ggcgcctaca acgtcaacat caagctggac atcacctccc acaacgagga ctacaccatc
      841 gtggaacagt acgagcgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
      901 aagtaaccaa ctttattata catagttgca ctcgagcctc tagaactata gtgagtcgta
      961 ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     1021 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     1081 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     1141 gggggaggtg tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt gggcccggta
     1201 cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     1261 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     1321 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     1381 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1441 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     1501 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1561 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1621 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1681 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1741 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1801 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1861 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1921 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1981 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2041 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     2101 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2161 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2221 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2281 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2341 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2401 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2461 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2521 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     2581 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     2641 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     2701 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     2761 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     2821 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     2881 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     2941 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3001 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3061 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3121 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3181 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3241 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3301 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3361 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     3421 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3481 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3541 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3601 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3661 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3721 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3781 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3841 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     3901 gatcggtgcg ggcctcttcg ctattacgcc agtcgaccat agccaattca atatggcgta
     3961 tatggactca tgccaattca atatggtgga tctggacctg tgccaattca atatggcgta
     4021 tatggactcg tgccaattca atatggtgga tctggacccc agccaattca atatggcgga
     4081 cttggcacca tgccaattca atatggcgga cttggcactg tgccaactgg ggaggggtct
     4141 acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc cgccatattg
     4201 aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc aatatatagg
     4261 caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata ctatgtattg
     4321 gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat agcccctccc
     4381 aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag ccactccccc
     4441 attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg gcggtcctat
     4501 tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg cctggctcaa
     4561 tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc tcattgccca
     4621 ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc cacttggcag
     4681 tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag gacgccaggg
     4741 tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag
     4801 tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt ccattgacgt
     4861 aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt ttagggaac
//
