LOCUS       sequence-274395-        4992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of DsRed-Express2 with nuclear localization 
            signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4992)
  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 4992)
  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-DsRed-cNLS
FEATURES             Location/Qualifiers
     source          1..4992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    86..134
                     /note="synthetic polyadenylation signal"
     misc_feature    148..239
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     188..207
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        243..610
                     /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    251..269
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    287..305
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    323..341
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    359..377
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    395..413
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    431..449
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    467..485
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      620..629
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             626..1300
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(806..826)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1024..1043
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1218..1241
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             1325..1345
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1349..1369
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1373..1393
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1565..1646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1679..1783
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1784..1852,1853..2644)
                     /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(2002..2021)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2815..3403
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3304..3323
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3557..3574
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3585..3666
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3783..4529)
                     /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(join(4548..4992,1..66))
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4611..4629)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
ORIGIN
        1 cgcccagagc agccgcgtcc ctgcgcaaac ccagggctgc cttggaaaag gcgcaacccc
       61 aaccccgtgg aattatcacc tcgagaataa aatatcttta ttttcattac atctgtgtgt
      121 tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca
      181 aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctctc
      241 tcgagtttac tccctatcag tgatagagaa cgtatgaaga gtttactccc tatcagtgat
      301 agagaacgta tgcagacttt actccctatc agtgatagag aacgtataag gagtttactc
      361 cctatcagtg atagagaacg tatgaccagt ttactcccta tcagtgatag agaacgtatc
      421 tacagtttac tccctatcag tgatagagaa cgtatatcca gtttactccc tatcagtgat
      481 agagaacgta taagctttgc ttatgtaaac cagggcgcct ataaaagagt gctgattttt
      541 tgagtaaact tcaattccac aacacttttg tcttatacca actttccgta ccacttccta
      601 ccctcgtaaa gaattcgccg ccaccatgga tagcactgag aacgtcatca agcccttcat
      661 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
      721 gggcgagggc aagccctacg agggcaccca gaccgccaag ctgcaggtga ccaagggcgg
      781 ccccctgccc ttcgcctggg acatcctgtc cccccagttc cagtacggct ccaaggtgta
      841 cgtgaagcac cccgccgaca tccccgacta caagaagctg tccttccccg agggcttcaa
      901 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
      961 cctgcaggac ggcaccttca tctaccacgt gaagttcatc ggcgtgaact tcccctccga
     1021 cggccccgta atgcagaaga agactctggg ctgggagccc tccaccgagc gcctgtaccc
     1081 ccgcgacggc gtgctgaagg gcgagatcca caaggcgctg aagctgaagg gcggcggcca
     1141 ctacctggtg gagttcaagt caatctacat ggccaagaag cccgtgaagc tgcccggcta
     1201 ctactacgtg gactccaagc tggacatcac ctcccacaac gaggactaca ccgtggtgga
     1261 gcagtacgag cgcgccgagg cccgccacca cctgttccag tccggactca gatctcgagc
     1321 tgatccaaaa aagaagagaa aggtagatcc aaaaaagaag agaaaggtag atccaaaaaa
     1381 gaagagaaag gtaggatcca ccggatctag ataactgatc aaccggtgga tccagatctc
     1441 tgcagcatat gtctagagga tcataatcag ccataccaca tttgtagagg ttttacttgc
     1501 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     1561 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     1621 cacaaataaa gcattttttt cactgccgac gtcaggtggc acttttcggg gaaatgtgcg
     1681 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1741 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     1801 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     1861 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     1921 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     1981 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     2041 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2101 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2161 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2221 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2281 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     2341 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2401 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2461 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     2521 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2581 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2641 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2701 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2761 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2821 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2881 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2941 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3001 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3061 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3121 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3181 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3241 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3301 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3361 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3421 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3481 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3541 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagct cggggcagtg aaaaaaatgc
     3601 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     3661 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     3721 gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcctctggag
     3781 atttacccgg ggagcatgtc aaggtcaaaa tcgtcaagag cgtcagcagg cagcatatca
     3841 aggtcaaagt cgtcaagggc atcggctggg agcatgtcta agtcaaaatc gtcaagggcg
     3901 tcggtcggcc cgccgctttc gcactttagc tgtttctcca ggccacatat gattagttcc
     3961 aggccgaaaa ggaaggcagg ttcggctccc tgccggtcga acagctcaat tgcttgtttc
     4021 agaagtgggg gcatagaatc ggtggtaggt gtctctcttt cctcttttgc tacttgatgc
     4081 tcctgttcct ccaatacgca gcccagtgta aagtggccca cggcggacag agcgtacagt
     4141 gcgttctcca gggagaagcc ttgctgacac aggaacgcga gctgattttc cagggtttcg
     4201 tactgtttct ctgttgggcg ggtgccgaga tgcactttag ccccgtcgcg atgtgagagg
     4261 agagcacagc ggtatgactt ggcgttgttc cgcagaaagt cttgccatga ctcgccttcc
     4321 agggggcagg agtgggtatg atgcctgtcc agcatctcga ttggcagggc atcgagcagg
     4381 gcccgcttgt tcttcacgtg ccagtacagg gtaggctgct caactcccag cttttgagcg
     4441 agtttccttg tcgtcaggcc ttcgataccg actccattga gtaattccag agcagagttt
     4501 atgactttgc tcttgtccag tctagacatg gtaattcgat gatccccctg gggagagagg
     4561 tcggtgattc ggtcaacgag ggagccgact gccgacgtgc gctccggagg cttgcagaat
     4621 gcggaacacc gcgcgggcag gaacagggcc cacactaccg ccccacaccc cgcctcccgc
     4681 accgcccctt cccggccgct gctctcggcg cgccctgctg agcagccgct attggccaca
     4741 gcccatcgcg gtcggcgcgc tgccattgct ccctggcgct gtccgtctgc gagggtacta
     4801 gtgagacgtg cggcttccgt ttgtcacgtc cggcacgccg cgaaccgcaa ggaaccttcc
     4861 cgacttaggg gcggagcagg aagcgtcgcc ggggggccca caagggtagc ggcgaagatc
     4921 cgggtgacgc tgcgaacgga cgtgaagaat gtgcgagacc cagggtcggc gccgctgcgt
     4981 ttcccggaac ca
//
