LOCUS       sequence-76199-e        4356 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of tdTomato under the control of the TRE-promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4356)
  AUTHORS   Tang JC, Szikra T, Kozorovitskiy Y, Teixiera M, Sabatini BL, Roska 
            B, Cepko CL
  TITLE     A nanobody-based system using fluorescent proteins as scaffolds for 
            cell-specific gene manipulation.
  JOURNAL   Cell. 2013 Aug 15;154(4):928-39. doi: 10.1016/j.cell.2013.07.021.
  PUBMED    23953120
REFERENCE   2  (bases 1 to 4356)
  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: 
            "10.1016/j.cell.2013.07"; journalName: "Cell"; date: "2013-08-15- 
            15"; volume: "154"; issue: "4"; pages: "928-39"
COMMENT     Sequence Label: pTRE-tdTomato
FEATURES             Location/Qualifiers
     source          1..4356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        109..423
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    117..135
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    153..171
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    188..206
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    224..242
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    260..278
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    295..313
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    331..349
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     396..420
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      434..443
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             440..1870
                     /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"
     polyA_signal    1910..2031
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1921..1940)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1975..1994
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2279..2296)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2450..3038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2530..2549)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3238..4029,4030..4098))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4203..4251
                     /note="synthetic polyadenylation signal"
     misc_feature    4265..4356
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4305..4324
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcctcgagga tatcaagatc tggcctcggc
       61 ggccaagctt gcatgcctgc aggtcacgag gccctttcgt cttcactcga gtttactccc
      121 tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag aacgatgtcg
      181 agtttactcc ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga
      241 gaacgtatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag tttatcccta
      301 tcagtgatag agaacgtatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga
      361 ggtaggcgtg tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc
      421 gcctggagaa ttcgccacca tggtgagcaa gggcgaggag gtcatcaaag agttcatgcg
      481 cttcaaggtg cgcatggagg gctccatgaa cggccacgag ttcgagatcg agggcgaggg
      541 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc
      601 cctgcccttc gcctgggaca tcctgtcccc ccagttcatg tacggctcca aggcgtacgt
      661 gaagcacccc gccgacatcc ccgattacaa gaagctgtcc ttccccgagg gcttcaagtg
      721 ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc gtgacccagg actcctccct
      781 gcaggacggc acgctgatct acaaggtgaa gatgcgcggc accaacttcc cccccgacgg
      841 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc accgagcgcc tgtacccccg
      901 cgacggcgtg ctgaagggcg agatccacca ggccctgaag ctgaaggacg gcggccacta
      961 cctggtggag ttcaagacca tctacatggc caagaagccc gtgcaactgc ccggctacta
     1021 ctacgtggac accaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     1081 gtacgagcgc tccgagggcc gccaccacct gttcctgggg catggcaccg gcagcaccgg
     1141 cagcggcagc tccggcaccg cctcctccga ggacaacaac atggccgtca tcaaagagtt
     1201 catgcgcttc aaggtgcgca tggagggctc catgaacggc cacgagttcg agatcgaggg
     1261 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     1321 cggccccctg cccttcgcct gggacatcct gtccccccag ttcatgtacg gctccaaggc
     1381 gtacgtgaag caccccgccg acatccccga ttacaagaag ctgtccttcc ccgagggctt
     1441 caagtgggag cgcgtgatga acttcgagga cggcggtctg gtgaccgtga cccaggactc
     1501 ctccctgcag gacggcacgc tgatctacaa ggtgaagatg cgcggcacca acttcccccc
     1561 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta
     1621 cccccgcgac ggcgtgctga agggcgagat ccaccaggcc ctgaagctga aggacggcgg
     1681 ccactacctg gtggagttca agaccatcta catggccaag aagcccgtgc aactgcccgg
     1741 ctactactac gtggacacca agctggacat cacctcccac aacgaggact acaccatcgt
     1801 ggaacagtac gagcgctccg agggccgcca ccacctgttc ctgtacggca tggacgagct
     1861 gtacaagtaa tctagagtcg gggcggccgg ccgcttcgag cagacatgat aagatacatt
     1921 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     1981 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2041 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     2101 taaaacctct acaaatgtgg taaaatcgat aaggatccgt cgaccgatgc ccttgagagc
     2161 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
     2221 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct
     2281 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2341 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2401 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2461 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2521 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2581 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2641 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2701 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2761 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2821 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2881 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2941 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3001 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3061 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3121 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3181 gtatatatga gtaaacttgg tctgacagcg gccgcaaatg ctaaaccact gcagtggtta
     3241 ccagtgcttg atcagtgagg caccgatctc agcgatctgc ctatttcgtt cgtccatagt
     3301 ggcctgactc cccgtcgtgt agatcactac gattcgtgag ggcttaccat caggccccag
     3361 cgcagcaatg atgccgcgag agccgcgttc accggccccc gatttgtcag caatgaacca
     3421 gccagcaggg agggccgagc gaagaagtgg tcctgctact ttgtccgcct ccatccagtc
     3481 tatgagctgc tgtcgtgatg ctagagtaag aagttcgcca gtgagtagtt tccgaagagt
     3541 tgtggccatt gctactggca tcgtggtatc acgctcgtcg ttcggtatgg cttcgttcaa
     3601 ctctggttcc cagcggtcaa gccgggtcac atgatcaccc atattatgaa gaaatgcagt
     3661 cagctcctta gggcctccga tcgttgtcag aagtaagttg gccgcggtgt tgtcgctcat
     3721 ggtaatggca gcactacaca attctcttac cgtcatgcca tccgtaagat gcttttccgt
     3781 gaccggcgag tactcaacca agtcgttttg tgagtagtgt atacggcgac caagctgctc
     3841 ttgcccggcg tctatacggg acaacaccgc gccacatagc agtactttga aagtgctcat
     3901 catcgggaat cgttcttcgg ggcggaaaga ctcaaggatc ttgccgctat tgagatccag
     3961 ttcgatatag cccactcttg cacccagttg atcttcagca tcttttactt tcaccagcgt
     4021 ttcggggtgt gcaaaaacag gcaagcaaaa tgccgcaaag aagggaatga gtgcgacacg
     4081 aaaatgttgg atgctcatac tcgtcctttt tcaatattat tgaagcattt atcagggtta
     4141 ctagtacgtc tctcaaggat aagtaagtaa tattaaggta cgggaggtat tggacaggcc
     4201 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
     4261 tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg
     4321 tccccagtgc aagtgcaggt gccagaacat ttctct
//
