LOCUS       sequence-274631-        3228 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DsRedT4 ORF with stop codon between attL1-attL2 on Gateway entry 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3228)
  TITLE     Minoru Ko Lab plasmids
REFERENCE   2  (bases 1 to 3228)
  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"
COMMENT     Sequence Label: pENTR-DsRedT4
FEATURES             Location/Qualifiers
     source          1..3228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             674..1351
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEPSTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(804..822)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(854..874)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1072..1091
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1266..1289
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(1353..1452)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1469..1488)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1470..1488)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1493..1509)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1493..1509)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1611..1630)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1622..2431
                     /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(1689..1708)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2578..3166
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3067..3086
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatggcct cctccgagga cgtcatcaag gagttcatgc gcttcaaggt
      721 gcgcatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
      781 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt
      841 cgcctgggac atcctgtccc cccagttcca gtacggctcc aaggtgtacg tgaagcaccc
      901 cgccgacatc cccgactaca agaagctgtc cttccccgag ggcttcaagt gggagcgcgt
      961 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     1021 ctgcttcatc tacaaggtga agttcatcgg cgtgaacttc ccctccgacg gccccgtaat
     1081 gcagaagaag actatgggct gggagccctc caccgagcgc ctgtaccccc gcgacggcgt
     1141 gctgaagggc gagatccaca aggccctgaa gctgaaggac ggcggccact acctggtgga
     1201 gttcaagtcc atctacatgg ccaagaagcc cgtgcagctg cccggctact actacgtgga
     1261 ctccaagctg gacatcacct cccacaacga ggactacacc atcgtggagc agtacgagcg
     1321 cgccgagggc cgccaccacc tgttcctgta gcacccagct ttcttgtaca aagttggcat
     1381 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     1441 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     1501 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     1561 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     1621 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     1681 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     1741 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     1801 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     1861 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     1921 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     1981 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     2041 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     2101 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2161 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     2221 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     2281 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     2341 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     2401 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     2461 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     2521 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2581 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2641 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2701 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2761 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2821 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2881 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2941 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3001 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3061 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3121 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3181 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
