LOCUS       sequence-200868-        3498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides linker:mCherry as GreenGate C-terminal tag module..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3498)
  AUTHORS   Lampropoulos A, Sutikovic Z, Wenzl C, Maegele I, Lohmann JU, Forner 
            J
  TITLE     GreenGate - A Novel, Versatile, and Efficient Cloning System for 
            Plant Transgenesis.
  JOURNAL   PLoS One. 2013 Dec 20;8(12):e83043. doi: 
            10.1371/journal.pone.0083043.
  PUBMED    24376629
REFERENCE   2  (bases 1 to 3498)
  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 One.";
            date: "2013-12-20"; volume: "8(12):e83043. doi"; pages: " 
            10.1371/journal.pone.0083043"
COMMENT     Sequence Label: pGGD003
FEATURES             Location/Qualifiers
     source          1..3498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    87..108
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(123..128,129..146,147..153)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    161..177
                     /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        191..209
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     191..208
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             332..1042
                     /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(477..495)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     745..764
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1063..1082)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1064..1082)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1092..1114)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1308..1327)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1427..1449
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1487..1505)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1573..1677
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1678..1746,1747..2538)
                     /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     complement(1896..1915)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2709..3297
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3198..3217
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3451..3468
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
       61 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
      121 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
      181 cacacgagct atttaggtga cactatagaa gtgaagcttg gtctcatcag gtgcagcggc
      241 cgcttcaggg agtggttccg gaagcggctc tggatcgggc tcagggagtg gttccggcag
      301 cggctctgga tcggcgatcg cagctgcagc catggtgagc aagggcgagg aggataacat
      361 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
      421 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
      481 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
      541 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
      601 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
      661 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
      721 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagacgatgg gctgggaggc
      781 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
      841 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
      901 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
      961 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     1021 catggacgag ctgtacaagt agctgctgag accgaattct cgccctatag tgagtcgtat
     1081 tacctggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     1141 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     1201 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     1261 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     1321 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     1381 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     1441 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     1501 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     1561 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     1621 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     1681 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     1741 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     1801 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     1861 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     1921 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     1981 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     2041 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     2101 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2161 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2221 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     2281 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2341 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggctctcga
     2401 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2461 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2521 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     2581 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     2641 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2701 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2761 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2821 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     2881 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2941 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3001 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3061 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3121 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3181 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3241 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3301 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3361 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3421 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3481 cgcccaatac gcaaaccg
//
