LOCUS       sequence-208135-        5463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tdTomato under the TRE promoter in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5463)
  AUTHORS   Sakaguchi R, Leiwe MN, Imai T
  TITLE     Bright multicolor labeling of neuronal circuits with fluorescent 
            proteins and chemical tags.
  JOURNAL   eLife. 2018 Nov 20;7. pii: 40350. doi: 10.7554/eLife.40350.
  PUBMED    30454553
REFERENCE   2  (bases 1 to 5463)
  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: "eLife."; 
            date: "2018-11-20"; volume: "7. pii: 40350. doi"; pages: " 
            10.7554/eLife.40350"
COMMENT     Sequence Label: pBS-TRE-tdTomato-WPRE
FEATURES             Location/Qualifiers
     source          1..5463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    659..677
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    695..713
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    730..748
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    766..784
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    802..820
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    837..855
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    873..891
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        903..941
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     938..962
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      976..985
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             982..2412
                     /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"
     misc_feature    2431..3019
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2484..2504)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2902..2913)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3180..3261
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3217..3236)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3275..3295)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3275..3293)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3314..3330)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3314..3330)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3327..3349)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3338..3354
                     /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(3362..3368,3369..3386,3387..3392))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3407..3428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3545..3562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3716..4304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3796..3815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4475..5266,5267..5335))
                     /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     5098..5117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5336..5440)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggtttactc
      661 cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgatgt
      721 cgagtttact ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata
      781 gagaacgtat gtcgagttta ctccctatca gtgatagaga acgtatgtcg agtttatccc
      841 tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc
      901 gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg aaccgtcaga
      961 tcgcctggag aattcgccac catggtgagc aagggcgagg aggtcatcaa agagttcatg
     1021 cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat cgagggcgag
     1081 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc
     1141 cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac
     1201 gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag
     1261 tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc
     1321 ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac
     1381 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc
     1441 cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccac
     1501 tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac
     1561 tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa
     1621 cagtacgagc gctccgaggg ccgccaccac ctgttcctgg ggcatggcac cggcagcacc
     1681 ggcagcggca gctccggcac cgcctcctcc gaggacaaca acatggccgt catcaaagag
     1741 ttcatgcgct tcaaggtgcg catggagggc tccatgaacg gccacgagtt cgagatcgag
     1801 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     1861 ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttcatgta cggctccaag
     1921 gcgtacgtga agcaccccgc cgacatcccc gattacaaga agctgtcctt ccccgagggc
     1981 ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc tggtgaccgt gacccaggac
     2041 tcctccctgc aggacggcac gctgatctac aaggtgaaga tgcgcggcac caacttcccc
     2101 cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg
     2161 tacccccgcg acggcgtgct gaagggcgag atccaccagg ccctgaagct gaaggacggc
     2221 ggccactacc tggtggagtt caagaccatc tacatggcca agaagcccgt gcaactgccc
     2281 ggctactact acgtggacac caagctggac atcacctccc acaacgagga ctacaccatc
     2341 gtggaacagt acgagcgctc cgagggccgc caccacctgt tcctgtacgg catggacgag
     2401 ctgtacaagt aagcggccgc gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     2461 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2521 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2581 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2641 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2701 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     2761 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     2821 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     2881 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     2941 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3001 ctttgggccg cctccccgca tcgataccgt cgacgcggcc gcatcgataa gcttgtcgac
     3061 gatatctcta gaggatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     3121 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     3181 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3241 ataaagcatt tttttcactg cctcgctttt gttcccttta gtgagggtta attgcgcgct
     3301 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3361 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3421 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3481 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3541 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3601 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3661 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3721 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3781 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3841 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3901 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3961 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4021 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4081 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4141 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4201 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4261 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4321 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4381 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4441 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4501 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4561 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4621 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4681 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4741 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4801 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4861 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4921 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4981 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5041 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5101 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5161 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5221 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5281 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5341 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5401 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5461 cac
//
