LOCUS       sequence-156007-        3984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway-compatible tdTomato fusion reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3984)
  AUTHORS   Fowler DK, Stewart S, Seredick S, Eisen JS, Stankunas K, Washbourne 
            P
  TITLE     A MultiSite Gateway Toolkit for Rapid Cloning of Vertebrate 
            Expression Constructs with Diverse Research Applications.
  JOURNAL   PLoS One. 2016 Aug 8;11(8):e0159277. doi: 
            10.1371/journal.pone.0159277. eCollection 2016.
  PUBMED    27500400
REFERENCE   2  (bases 1 to 3984)
  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: "PLoS One.";
            date: "2016-08-8"; volume: "11(8):e0159277. doi"; pages: " 
            10.1371/journal.pone.0159277. eCollection 2016"
COMMENT     Sequence Label: pME-tdTomato no stop
FEATURES             Location/Qualifiers
     source          1..3984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      23..32
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             29..1456
                     /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"
     protein_bind    complement(1458..1557)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1574..1593)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1575..1593)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1598..1614)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1598..1614)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1716..1735)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1727..2536
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1794..1813)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2683..3271
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3172..3191
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3425..3442
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3601..3628)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3720..3806)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3855..3877
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3869..3886
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3870..3886
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    join(3902..3984,1..17)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 gtacaaaaaa gcaggctccg ccgccaccat ggtgagcaag ggcgaggagg tcatcaaaga
       61 gttcatgcgc ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga
      121 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
      181 gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa
      241 ggcgtacgtg aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg
      301 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga
      361 ctcctccctg caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc
      421 ccccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct
      481 gtacccccgc gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg
      541 cggccactac ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc
      601 cggctactac tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat
      661 cgtggaacag tacgagcgct ccgagggccg ccaccacctg ttcctggggc atggcaccgg
      721 cagcaccggc agcggcagct ccggcaccgc ctcctccgag gacaacaaca tggccgtcat
      781 caaagagttc atgcgcttca aggtgcgcat ggagggctcc atgaacggcc acgagttcga
      841 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
      901 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt tcatgtacgg
      961 ctccaaggcg tacgtgaagc accccgccga catccccgat tacaagaagc tgtccttccc
     1021 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac
     1081 ccaggactcc tccctgcagg acggcacgct gatctacaag gtgaagatgc gcggcaccaa
     1141 cttccccccc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga
     1201 gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa
     1261 ggacggcggc cactacctgg tggagttcaa gaccatctac atggccaaga agcccgtgca
     1321 actgcccggc tactactacg tggacaccaa gctggacatc acctcccaca acgaggacta
     1381 caccatcgtg gaacagtacg agcgctccga gggccgccac cacctgttcc tgtacggcat
     1441 ggacgagctg tacaaggacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     1501 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     1561 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     1621 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     1681 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     1741 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     1801 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     1861 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     1921 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     1981 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2041 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2101 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2161 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2221 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     2281 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2341 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     2401 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     2461 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     2521 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     2581 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     2641 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2701 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2761 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2821 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2881 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2941 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3001 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3061 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3121 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3181 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3241 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3301 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3361 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3421 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     3481 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     3541 cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca
     3601 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg
     3661 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
     3721 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     3781 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct
     3841 agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc
     3901 ccaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgatgagca
     3961 atgctttttt ataatgccaa cttt
//
