LOCUS       sequence-258690-        3387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains the sequence of mKate2 for C-term fusions in the pGGD000 
            backbone (GreenGate).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3387)
  AUTHORS   Denay G, Schultz P, Hansch S, Weidtkamp-Peters S, Simon R
  TITLE     Over the rainbow: A practical guide for fluorescent protein 
            selection in plant FRET experiments.
  JOURNAL   Plant Direct. 2019 Dec 6;3(12):e00189. doi: 10.1002/pld3.189. 
            eCollection 2019 Dec.
  PUBMED    31844834
REFERENCE   2  (bases 1 to 3387)
  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: "Plant 
            Direct."; date: "2019-12-6"; volume: "3(12):e00189. doi"; pages: " 
            10.1002/pld3.189. eCollection 2019 Dec"
COMMENT     Sequence Label: pRD141
FEATURES             Location/Qualifiers
     source          1..3387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(15..20,21..38,39..45)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    53..69
                     /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        83..101
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     83..100
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(844..863)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(845..863)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(873..895)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1089..1108)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1208..1230
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1268..1286)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1354..1458
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1459..1527,1528..2319)
                     /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(1677..1696)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2490..3078
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2979..2998
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3232..3249
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3366..3387
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
       61 ggataacaat ttcacacgag ctatttaggt gacactatag aagtgaagct tggtctcatc
      121 agttatggtg tcggagctca tcaaggagaa catgcacatg aagctctaca tggagggcac
      181 cgtcaacaac caccacttca agtgcacctc ggaaggcgag ggcaagccct acgagggcac
      241 ccagaccatg cgcatcaagg ccgtcgaggg tggtcctctg cccttcgcct tcgacatcct
      301 cgccacctcg ttcatgtacg gctccaagac gttcatcaac cacacgcagg gcatccccga
      361 ctttttcaag cagtcgttcc ccgagggctt cacctgggag cgtgtcacca cctacgagga
      421 cggcggtgtc ctcaccgcca cccaggacac ctcgctccag gacggctgcc tcatctacaa
      481 cgtcaagatc cgcggtgtca acttcccctc gaacggtccc gtcatgcaga agaagacgct
      541 cggctgggag gcctcgaccg agacgctcta ccccgccgac ggtggcctcg agggtcgtgc
      601 cgacatggcg ctcaagctcg tcggtggtgg tcacctcatc tgcaacctca agaccaccta
      661 ccgctccaag aagcccgcca agaacctcaa gatgcccggt gtctactacg tcgaccgtcg
      721 tctcgagcgc atcaaggagg ccgacaagga aacctacgtc gagcagcacg aggtcgccgt
      781 cgctcgctac tgcgatctgc cctcgaaact cggtcaccgc taactgctga gaccgaattc
      841 tcgccctata gtgagtcgta ttacctggcc gtcgttttac aacgtcgtga ctgggaaaac
      901 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
      961 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     1021 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     1081 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     1141 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     1201 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     1261 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     1321 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1381 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1441 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1501 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1561 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1621 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1681 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     1741 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1801 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1861 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1921 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     1981 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     2041 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2101 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2161 gccggtgagc gtggctctcg aggtatcatt gcagcactgg ggccagatgg taagccctcc
     2221 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2281 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2341 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2401 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2461 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2521 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2581 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2641 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2701 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2761 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2821 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2881 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2941 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3001 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3061 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3121 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3181 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3241 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     3301 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     3361 gcaattaatg tgagttagct cactcat
//
