LOCUS       sequence-274730-        3303 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mCherry ORF with stop codon between attL5-attL2 on Gateway entry 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3303)
  TITLE     Minoru Ko Lab plasmids
REFERENCE   2  (bases 1 to 3303)
  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-L5L2-mCherry-stop
FEATURES             Location/Qualifiers
     source          1..3303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     95..112
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(271..298)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(390..476)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     525..547
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     539..556
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     540..556
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    573..668
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             677..1384
                     /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(822..840)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1090..1109
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(1418..1517)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1547..1566)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1548..1566)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1571..1587)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1571..1587)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1689..1708)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1700..2509
                     /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(1767..1786)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2656..3244
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3145..3164
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
       61 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      121 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
      181 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac
      241 cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc
      301 ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc
      361 ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac
      421 aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc
      481 ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg
      541 taaaacgacg gccagtctta agctcgggcc ccaaataatg attttatttt gactgatagt
      601 gacctgttcg ttgcaacaaa ttgatgagca atgctttttt ataatgccaa ctttgtatac
      661 aaaagttgta ggcaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
      721 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
      781 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
      841 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
      901 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
      961 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     1021 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     1081 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     1141 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     1201 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     1261 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     1321 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     1381 caagggatat ccatcacact ggcggccgct cgagtaaacc cagctttctt gtacaaagtt
     1441 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     1501 aataaaatca ttatttgcca tccagctgca gggcggccgc gatatcccct atagtgagtc
     1561 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     1621 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     1681 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     1741 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     1801 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     1861 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     1921 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     1981 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2041 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2101 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     2161 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     2221 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     2281 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     2341 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     2401 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     2461 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     2521 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     2581 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     2641 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2701 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2761 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2821 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2881 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2941 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3001 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3061 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3121 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3181 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3241 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3301 cat
//
