LOCUS       sequence-274399-        5025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of mCherry with nuclear localization signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5025)
  AUTHORS   Taylor-Weiner H, Grigsby CL, Ferreira DMS, Dias JM, Stevens MM, Ruas
            JL, Teixeira AI
  TITLE     Modeling the transport of nuclear proteins along single skeletal 
            muscle cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Feb 11;117(6):2978-2986. doi: 
            10.1073/pnas.1919600117. Epub 2020 Jan 27.
  PUBMED    31988126
REFERENCE   2  (bases 1 to 5025)
  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"; doi: 
            "10.1073/pnas.1919600117"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2020-02-11- 11"; volume: "117"; issue: "6"; pages: 
            "2978-2986"
COMMENT     Sequence Label: pTetOne-mCherry-cNLS
FEATURES             Location/Qualifiers
     source          1..5025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..371
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    12..30
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    48..66
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    84..102
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    120..138
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    156..174
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    192..210
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    228..246
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      381..390
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             387..1094
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(532..550)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     800..819
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1119..1139
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1143..1163
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1167..1187
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1359..1440
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1473..1577
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1578..1646,1647..2438)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1796..1815)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2609..3197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3098..3117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3351..3368
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3379..3460
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3577..4323)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     promoter        complement(4342..4852)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4405..4423)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     polyA_signal    4872..4920
                     /note="synthetic polyadenylation signal"
     misc_feature    4934..5025
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4974..4993
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ctcgagttta ctccctatca gtgatagaga acgtatgaag agtttactcc ctatcagtga
       61 tagagaacgt atgcagactt tactccctat cagtgataga gaacgtataa ggagtttact
      121 ccctatcagt gatagagaac gtatgaccag tttactccct atcagtgata gagaacgtat
      181 ctacagttta ctccctatca gtgatagaga acgtatatcc agtttactcc ctatcagtga
      241 tagagaacgt ataagctttg cttatgtaaa ccagggcgcc tataaaagag tgctgatttt
      301 ttgagtaaac ttcaattcca caacactttt gtcttatacc aactttccgt accacttcct
      361 accctcgtaa agaattcgcc gccaccatgg tgagcaaggg cgaggaggat aacatggcca
      421 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
      481 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
      541 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
      601 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
      661 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
      721 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
      781 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
      841 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
      901 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
      961 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     1021 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     1081 acgagctgta caagtccgga ctcagatctc gagctgatcc aaaaaagaag agaaaggtag
     1141 atccaaaaaa gaagagaaag gtagatccaa aaaagaagag aaaggtagga tccaccggat
     1201 ctagataact gatcaaccgg tggatccaga tctctgcagc atatgtctag aggatcataa
     1261 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     1321 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     1381 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1441 cgacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1501 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1561 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1621 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1681 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1741 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1801 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     1861 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1921 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1981 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2041 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2101 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2161 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2221 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2281 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2341 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2401 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2461 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2521 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2581 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2641 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2701 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2761 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2821 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2881 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2941 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3001 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3061 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3121 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3181 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3241 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3301 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3361 tgagcgagga agctcggggc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3421 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3481 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     3541 caaatgtggt atggctgatt atgatcctct ggagatttac ccggggagca tgtcaaggtc
     3601 aaaatcgtca agagcgtcag caggcagcat atcaaggtca aagtcgtcaa gggcatcggc
     3661 tgggagcatg tctaagtcaa aatcgtcaag ggcgtcggtc ggcccgccgc tttcgcactt
     3721 tagctgtttc tccaggccac atatgattag ttccaggccg aaaaggaagg caggttcggc
     3781 tccctgccgg tcgaacagct caattgcttg tttcagaagt gggggcatag aatcggtggt
     3841 aggtgtctct ctttcctctt ttgctacttg atgctcctgt tcctccaata cgcagcccag
     3901 tgtaaagtgg cccacggcgg acagagcgta cagtgcgttc tccagggaga agccttgctg
     3961 acacaggaac gcgagctgat tttccagggt ttcgtactgt ttctctgttg ggcgggtgcc
     4021 gagatgcact ttagccccgt cgcgatgtga gaggagagca cagcggtatg acttggcgtt
     4081 gttccgcaga aagtcttgcc atgactcgcc ttccaggggg caggagtggg tatgatgcct
     4141 gtccagcatc tcgattggca gggcatcgag cagggcccgc ttgttcttca cgtgccagta
     4201 cagggtaggc tgctcaactc ccagcttttg agcgagtttc cttgtcgtca ggccttcgat
     4261 accgactcca ttgagtaatt ccagagcaga gtttatgact ttgctcttgt ccagtctaga
     4321 catggtaatt cgatgatccc cctggggaga gaggtcggtg attcggtcaa cgagggagcc
     4381 gactgccgac gtgcgctccg gaggcttgca gaatgcggaa caccgcgcgg gcaggaacag
     4441 ggcccacact accgccccac accccgcctc ccgcaccgcc ccttcccggc cgctgctctc
     4501 ggcgcgccct gctgagcagc cgctattggc cacagcccat cgcggtcggc gcgctgccat
     4561 tgctccctgg cgctgtccgt ctgcgagggt actagtgaga cgtgcggctt ccgtttgtca
     4621 cgtccggcac gccgcgaacc gcaaggaacc ttcccgactt aggggcggag caggaagcgt
     4681 cgccgggggg cccacaaggg tagcggcgaa gatccgggtg acgctgcgaa cggacgtgaa
     4741 gaatgtgcga gacccagggt cggcgccgct gcgtttcccg gaaccacgcc cagagcagcc
     4801 gcgtccctgc gcaaacccag ggctgccttg gaaaaggcgc aaccccaacc ccgtggaatt
     4861 atcacctcga gaataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg
     4921 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
     4981 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctct
//
