LOCUS       sequence-274727-        3311 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mCherry 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 3311)
  TITLE     Minoru Ko Lab plasmids
REFERENCE   2  (bases 1 to 3311)
  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_mCherry
FEATURES             Location/Qualifiers
     source          1..3311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      38..47
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             44..751
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(189..207)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     457..476
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(785..884)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(901..920)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(902..920)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(925..941)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(925..941)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1043..1062)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1054..1863
                     /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(1121..1140)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2010..2598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2499..2518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2752..2769
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2928..2955)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3047..3133)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3182..3204
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3196..3213
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3197..3213
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    join(3229..3311,1..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 gcaggctgct accggtctcg agaattcgcc accatggtga gcaagggcga
       61 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
      121 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
      181 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
      241 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
      301 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
      361 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
      421 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
      481 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
      541 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
      601 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
      661 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
      721 ctccaccggc ggcatggacg agctgtacaa gggatatcca tcacactggc ggccgctcga
      781 gtaaacccag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
      841 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc
      901 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
      961 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     1021 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     1081 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     1141 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     1201 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     1261 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     1321 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     1381 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     1441 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     1501 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     1561 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     1621 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     1681 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     1741 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     1801 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     1861 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     1921 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     1981 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2041 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2101 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2161 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2221 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2281 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2341 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2401 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2461 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2521 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2581 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2641 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     2701 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     2761 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     2821 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     2881 gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg
     2941 tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc
     3001 accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca
     3061 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct
     3121 ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt
     3181 tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggcccca aataatgatt
     3241 ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg cttttttata
     3301 atgccaactt t
//
