LOCUS       sequence-274407-        5745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of tdTomato with nuclear localization signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5745)
  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 5745)
  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-tdTomato-cNLS
FEATURES             Location/Qualifiers
     source          1..5745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      10..19
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             16..1443
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             1468..1488
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1492..1512
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1516..1536
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1708..1789
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1822..1926
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1927..1995,1996..2787)
                     /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(2145..2164)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2958..3546
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3447..3466
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3700..3717
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3728..3809
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3926..4672)
                     /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(4691..5201)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4754..4772)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     polyA_signal    5221..5269
                     /note="synthetic polyadenylation signal"
     misc_feature    5283..5374
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5323..5342
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        5378..5745
                     /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    5386..5404
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5422..5440
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5458..5476
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5494..5512
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5530..5548
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5566..5584
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5602..5620
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 gaattcgccg ccaccatggt gagcaagggc gaggaggtca tcaaagagtt catgcgcttc
       61 aaggtgcgca tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag
      121 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg
      181 cccttcgcct gggacatcct gtccccccag ttcatgtacg gctccaaggc gtacgtgaag
      241 caccccgccg acatccccga ttacaagaag ctgtccttcc ccgagggctt caagtgggag
      301 cgcgtgatga acttcgagga cggcggtctg gtgaccgtga cccaggactc ctccctgcag
      361 gacggcacgc tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc
      421 gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac
      481 ggcgtgctga agggcgagat ccaccaggcc ctgaagctga aggacggcgg ccactacctg
      541 gtggagttca agaccatcta catggccaag aagcccgtgc aactgcccgg ctactactac
      601 gtggacacca agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
      661 gagcgctccg agggccgcca ccacctgttc ctggggcatg gcaccggcag caccggcagc
      721 ggcagctccg gcaccgcctc ctccgaggac aacaacatgg ccgtcatcaa agagttcatg
      781 cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat cgagggcgag
      841 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc
      901 cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac
      961 gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag
     1021 tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc
     1081 ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac
     1141 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc
     1201 cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccac
     1261 tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac
     1321 tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa
     1381 cagtacgagc gctccgaggg ccgccaccac ctgttcctgt acggcatgga cgagctgtac
     1441 aagtccggac tcagatctcg agctgatcca aaaaagaaga gaaaggtaga tccaaaaaag
     1501 aagagaaagg tagatccaaa aaagaagaga aaggtaggat ccaccggatc tagataactg
     1561 atcaaccggt ggatccagat ctctgcagca tatgtctaga ggatcataat cagccatacc
     1621 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     1681 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     1741 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgcc gacgtcaggt
     1801 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1861 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1921 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1981 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     2041 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     2101 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     2161 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     2221 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     2281 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     2341 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     2401 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     2461 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     2521 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     2581 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     2641 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     2701 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     2761 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     2821 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     2881 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2941 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3001 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3061 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     3121 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3181 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3241 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3301 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3361 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3421 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3481 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3541 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3601 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3661 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3721 gctcggggca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3781 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3841 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     3901 tggctgatta tgatcctctg gagatttacc cggggagcat gtcaaggtca aaatcgtcaa
     3961 gagcgtcagc aggcagcata tcaaggtcaa agtcgtcaag ggcatcggct gggagcatgt
     4021 ctaagtcaaa atcgtcaagg gcgtcggtcg gcccgccgct ttcgcacttt agctgtttct
     4081 ccaggccaca tatgattagt tccaggccga aaaggaaggc aggttcggct ccctgccggt
     4141 cgaacagctc aattgcttgt ttcagaagtg ggggcataga atcggtggta ggtgtctctc
     4201 tttcctcttt tgctacttga tgctcctgtt cctccaatac gcagcccagt gtaaagtggc
     4261 ccacggcgga cagagcgtac agtgcgttct ccagggagaa gccttgctga cacaggaacg
     4321 cgagctgatt ttccagggtt tcgtactgtt tctctgttgg gcgggtgccg agatgcactt
     4381 tagccccgtc gcgatgtgag aggagagcac agcggtatga cttggcgttg ttccgcagaa
     4441 agtcttgcca tgactcgcct tccagggggc aggagtgggt atgatgcctg tccagcatct
     4501 cgattggcag ggcatcgagc agggcccgct tgttcttcac gtgccagtac agggtaggct
     4561 gctcaactcc cagcttttga gcgagtttcc ttgtcgtcag gccttcgata ccgactccat
     4621 tgagtaattc cagagcagag tttatgactt tgctcttgtc cagtctagac atggtaattc
     4681 gatgatcccc ctggggagag aggtcggtga ttcggtcaac gagggagccg actgccgacg
     4741 tgcgctccgg aggcttgcag aatgcggaac accgcgcggg caggaacagg gcccacacta
     4801 ccgccccaca ccccgcctcc cgcaccgccc cttcccggcc gctgctctcg gcgcgccctg
     4861 ctgagcagcc gctattggcc acagcccatc gcggtcggcg cgctgccatt gctccctggc
     4921 gctgtccgtc tgcgagggta ctagtgagac gtgcggcttc cgtttgtcac gtccggcacg
     4981 ccgcgaaccg caaggaacct tcccgactta ggggcggagc aggaagcgtc gccggggggc
     5041 ccacaagggt agcggcgaag atccgggtga cgctgcgaac ggacgtgaag aatgtgcgag
     5101 acccagggtc ggcgccgctg cgtttcccgg aaccacgccc agagcagccg cgtccctgcg
     5161 caaacccagg gctgccttgg aaaaggcgca accccaaccc cgtggaatta tcacctcgag
     5221 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
     5281 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
     5341 cccagtgcaa gtgcaggtgc cagaacattt ctctctcgag tttactccct atcagtgata
     5401 gagaacgtat gaagagttta ctccctatca gtgatagaga acgtatgcag actttactcc
     5461 ctatcagtga tagagaacgt ataaggagtt tactccctat cagtgataga gaacgtatga
     5521 ccagtttact ccctatcagt gatagagaac gtatctacag tttactccct atcagtgata
     5581 gagaacgtat atccagttta ctccctatca gtgatagaga acgtataagc tttgcttatg
     5641 taaaccaggg cgcctataaa agagtgctga ttttttgagt aaacttcaat tccacaacac
     5701 ttttgtctta taccaacttt ccgtaccact tcctaccctc gtaaa
//
