LOCUS       sequence-209940-        3350 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3350)
  AUTHORS   Ariotti N, Rae J, Giles N, Martel N, Sierecki E, Gambin Y, Hall TE, 
            Parton RG
  TITLE     Ultrastructural localisation of protein interactions using 
            conditionally stable nanobodies.
  JOURNAL   PLoS Biol. 2018 Apr 5;16(4):e2005473. doi: 
            10.1371/journal.pbio.2005473. eCollection 2018 Apr.
  PUBMED    29621251
REFERENCE   2  (bases 1 to 3350)
  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 
            Biol."; date: "2018-04-5"; volume: "16(4):e2005473. doi"; pages: " 
            10.1371/journal.pbio.2005473. eCollection 2018 Apr"
COMMENT     Sequence Label: p3E-mCherry
FEATURES             Location/Qualifiers
     source          1..3350
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     84..101
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(260..287)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(379..465)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     514..536
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     528..545
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     529..545
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(616..739)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             742..1449
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(884..902)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1152..1171
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1346..1369
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(1451..1546)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1583..1602)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1584..1602)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1607..1623)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1607..1623)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1725..1744)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1736..2545
                     /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(1803..1822)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2692..3280
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3181..3200
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
       61 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
      121 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
      181 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg
      241 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg
      301 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt
      361 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
      421 aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc
      481 ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt aaaacgacgg
      541 ccagtcttaa gctcgggccc tgcagctcta gagctcgaat tctacaggtc actaatacca
      601 tctaagtagt tggttcatag tgactgcata tgttgtgttt tacagtatta tgtagtctgt
      661 tttttatgca aaatctaatt taatatattg atatttatat cattttacgt ttctcgttca
      721 actttcttgt acaaagtggc tgtgagcaag ggcgaggagg acaacatggc catcatcaag
      781 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
      841 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
      901 aagggcggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
      961 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
     1021 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     1081 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     1141 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     1201 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     1261 ggcggccact acgacgccga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     1321 cccggcgcct acaacgtcaa catcaagctg gacatcacct cccacaacga ggactacacc
     1381 atcgtggaac agtacgagcg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     1441 tacaagtaac caactttatt atacaaagtt ggcattataa aaaagcattg cttatcaatt
     1501 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttggag ctccatggta
     1561 gcgttaacgc ggccgcgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     1621 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     1681 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     1741 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     1801 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     1861 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     1921 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     1981 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     2041 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2101 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2161 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     2221 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     2281 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     2341 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     2401 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     2461 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     2521 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     2581 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     2641 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2701 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2761 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2821 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2881 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2941 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3001 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3061 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3121 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3181 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3241 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3301 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
//
