LOCUS       sequence-234044-        3927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For making C-terminal mCherry fusions using multisite Gateway 
            cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3927)
  AUTHORS   Gu Y, Innes RW
  TITLE     The KEEP ON GOING protein of Arabidopsis recruits the ENHANCED 
            DISEASE RESISTANCE1 protein to trans-Golgi network/early endosome 
            vesicles.
  JOURNAL   Plant Physiol. 2011 Apr;155(4):1827-38. doi: 10.1104/pp.110.171785. 
            Epub 2011 Feb 22.
  PUBMED    21343429
REFERENCE   2  (bases 1 to 3927)
  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"; doi: 
            "10.1104/pp.110.171785"; journalName: "Plant Physiol"; date: 
            "2011-04"; volume: "155"; issue: "4"; pages: "1827-38"
COMMENT     Sequence Label: mCherry pBSDONR P4r-P2
FEATURES             Location/Qualifiers
     source          1..3927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     100..122
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     114..131
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     115..131
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     141..160
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        141..159
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(214..337)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             340..1050
                     /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(485..503)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     753..772
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(1052..1151)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1200..1216)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1200..1216)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1253..1273)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1253..1271)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1292..1308)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1292..1308)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1305..1327)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1316..1332
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1340..1346,1347..1364,1365..1370))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1385..1406
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1523..1540)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1694..2282)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1774..1793)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2453..3244,3245..3313))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3076..3095
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3314..3418)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3445..3901
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3532..3551)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3742..3763
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
       61 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
      121 cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc
      181 tacaggtcac taataccatc taagtagttg attcatagtg actggatatg ttgtgtttta
      241 cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca
      301 ttttacgttt ctcgttcaac ttttctatac aaagttgaca tggtgagcaa gggcgaggag
      361 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
      421 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
      481 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
      541 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
      601 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
      661 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
      721 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
      781 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
      841 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
      901 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
      961 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     1021 accggcggca tggacgagct gtacaagtaa tacccagctt tcttgtacaa agttggcatt
     1081 ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     1141 atcattattt gccatccagc tgcagggcgg ccgcgatatc gaattcctgc agcccggggg
     1201 atccactagt tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt
     1261 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1321 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1381 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1441 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1501 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1561 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1621 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1681 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1741 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1801 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1861 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1921 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1981 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2041 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2101 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2161 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2221 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2281 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2341 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2401 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2461 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2521 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2581 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2641 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2701 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2761 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     2821 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     2881 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     2941 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3001 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3061 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3121 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3181 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3241 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3301 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3361 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3421 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3481 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3541 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3601 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3661 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3721 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3781 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3841 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     3901 tccattcgcc attcaggctg cgcaact
//
