LOCUS       sequence-258695-        3399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains the sequence of mOrange 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 3399)
  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 3399)
  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: pGD254
FEATURES             Location/Qualifiers
     source          1..3399
                     /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"
     CDS             125..835
                     /codon_start=1
                     /product="orange monomeric derivative of DsRed fluorescent 
                     protein (Shanet et al., 2004)"
                     /label=mOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     538..557
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(856..875)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(857..875)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(885..907)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1101..1120)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1220..1242
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1280..1298)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1366..1470
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1471..1539,1540..2331)
                     /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(1689..1708)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2502..3090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2991..3010
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3244..3261
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3378..3399
                     /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 aggaatggtg agcaagggcg aggagaataa catggccatc atcaaggagt tcatgcgctt
      181 caaggtgcgc atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
      241 gggccgcccc tacgagggct ttcagaccgc taagctgaag gtgaccaagg gtggccccct
      301 gcccttcgcc tgggacatcc tgtcccctca gttcacctac ggctccaagg cctacgtgaa
      361 gcaccccgcc gacatccccg actacttcaa gctgtccttc cccgagggct tcaagtggga
      421 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
      481 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
      541 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
      601 cggcgccctg aagggcgaga tcaagatgag gctgaagctg aaggacggcg gccactacac
      661 ctccgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacat
      721 cgtcggcatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
      781 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtaactgct
      841 gagaccgaat tctcgcccta tagtgagtcg tattacctgg ccgtcgtttt acaacgtcgt
      901 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
      961 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     1021 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     1081 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     1141 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     1201 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     1261 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     1321 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1381 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1441 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1501 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1561 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1621 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1681 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     1741 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1801 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1861 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     1921 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     1981 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     2041 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2101 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2161 gataaatctg gagccggtga gcgtggctct cgaggtatca ttgcagcact ggggccagat
     2221 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2281 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2341 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2401 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2461 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2521 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2581 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2641 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2701 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2761 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2821 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2881 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2941 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3001 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3061 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3121 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3181 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3241 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     3301 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     3361 agtgagcgca acgcaattaa tgtgagttag ctcactcat
//
